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Soderini A, Depietri V, Crespe M, Rodriguez Y, Aragona A. The role of sentinel lymph node mapping in endometrial carcinoma. ACTA ACUST UNITED AC 2020; 72:367-383. [PMID: 32921021 DOI: 10.23736/s0026-4784.20.04626-2] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
Abstract
Endometrial cancer is the most commonly diagnosed gynecological malignancy in developing countries, and the second malignancy after cervical cancer in developing countries. The primary treatment is based on surgical and pathologic staging including extrafascial type A radical hysterectomy bilateral salpingo-oophorectomy and pelvic and latero-aortic lymphadenectomy. Minimally invasive surgery is the most widely used technique. Sentinel node biopsy is part of this concept and has reached the management of endometrial cancer. The aim of this review was to describe the history, the different injection techniques and results of sentinel node biopsy, and analyze the future role of this technique in endometrial carcinoma.
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Affiliation(s)
- Alejandro Soderini
- Unit of Gynecologic Oncology, Marie Curie Oncologic Hospital, University of Buenos Aires, Buenos Aires, Argentina -
| | - Valeria Depietri
- Unit of Gynecologic Oncology, Marie Curie Oncologic Hospital, University of Buenos Aires, Buenos Aires, Argentina
| | - Martin Crespe
- Unit of Gynecologic Oncology, Marie Curie Oncologic Hospital, University of Buenos Aires, Buenos Aires, Argentina
| | - Yanina Rodriguez
- Unit of Gynecologic Oncology, Marie Curie Oncologic Hospital, University of Buenos Aires, Buenos Aires, Argentina
| | - Alejandro Aragona
- Unit of Gynecologic Oncology, Marie Curie Oncologic Hospital, University of Buenos Aires, Buenos Aires, Argentina
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Lee GW, Park JY, Kim DY, Suh DS, Kim JH, Kim YM, Kim YT, Nam JH. Usefulness of sentinel lymph node mapping using indocyanine green and fluorescent imaging in the diagnosis of lymph node metastasis in endometrial cancer. J OBSTET GYNAECOL 2020; 41:605-611. [PMID: 32815448 DOI: 10.1080/01443615.2020.1787965] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/24/2022]
Abstract
The lymph node status is the most important prognostic factor for endometrial cancer. This study aimed to assess whether sentinel lymph node mapping (SLNM) is applicable in endometrial cancer. A retrospective review of patients with endometrial cancer who were diagnosed and treated in Asan Medical Centre from September 2015 to December 2017 was conducted. One hundred patients underwent robotic (da Vinci®) or laparoscopic surgical treatment, including SLNM with indocyanine green (ICG) fluorescence detection using the Firefly® and NIR/ICG systems. At least one lymph node area was observed in 100% of SLNM cases. Sentinel node detection and frozen biopsy were performed in all cases, and all patients with metastasis were found on SLNM. The sensitivity and negative predictive value were both 100% in the patient-by-patient and station-by-station analyses. SLNM appears to be a feasible method to reduce the morbidity and increase the detection rate in early-stage endometrial carcinoma.What is already known on this subject? There are studies that it is safe to diagnose the possibility of lymph node metastasis through sentinel lymph node mapping in endometrial cancer.What do the results of this study add? In this study, it is shown that the accuracy of sentinel lymph node mapping is 100% accurate.What are the implications of these findings for clinical practise and/or further research? Therefore, total lymphadenectomy will not be necessary for the future.
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Affiliation(s)
- Geon-Woo Lee
- Department of Obstetrics and Gynecology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea
| | - Jeong-Yeol Park
- Department of Pathology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea
| | - Dae-Yeon Kim
- Department of Pathology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea
| | - Dae-Shik Suh
- Department of Pathology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea
| | - Jong-Hyeok Kim
- Department of Pathology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea
| | - Yong-Man Kim
- Department of Pathology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea
| | - Young-Tak Kim
- Department of Pathology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea
| | - Joo-Hyun Nam
- Department of Obstetrics and Gynecology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea
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Oncologic impact of micrometastases or isolated tumor cells in sentinel lymph nodes of patients with endometrial cancer: a meta-analysis. Clin Transl Oncol 2019; 22:1272-1279. [PMID: 31863354 DOI: 10.1007/s12094-019-02249-x] [Citation(s) in RCA: 26] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/13/2019] [Accepted: 11/24/2019] [Indexed: 12/17/2022]
Abstract
PURPOSE There is a gap in knowledge regarding the impact of micrometastases (MIC) and isolated tumor cells (ITCs) found in the sentinel lymph nodes of patients with endometrial cancer. Here, we present a meta-analysis of the published literature on the rate of MIC and ITCs after lymphatic mapping and determine trends in postoperative management. METHODS Literature search of Medline and PubMed was done using the terms: micrometastases, isolated tumor cells, endometrial cancer, and sentinel lymph node. Inclusion criteria were: English-language manuscripts, retrospectives, or prospective studies published between January 1999 and June 2019. We removed manuscripts on sentinel node mapping that did not specify information on micrometastases or isolated tumor cells, non-English-language articles, no data about oncologic outcomes, and articles limited to ten cases or less. RESULTS A total of 45 manuscripts were reviewed, and 8 studies met inclusion criteria. We found that the total number of patients with MIC/ITCs was 286 (187 and 99, respectively). The 72% of patients detected with MIC/ITCs in sentinel nodes received adjuvant therapies. The MIC/ITCs group has a higher relative risk of recurrence of 1.34 (1.07, 1.67) than the negative group, even if the adjuvant therapy was given. CONCLUSION We noted that there is an increased relative risk of recurrence in patients with low-volume metastases, even after receiving adjuvant therapy. Whether adjuvant therapy is indicated remains a topic of debate because there are other uterine factors implicated in the prognosis. Multi-institutional tumor registries may help shed light on this important question.
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Accuracy of One-Step Nucleic Acid Amplification in Detecting Lymph Node Metastases in Endometrial Cancer. Pathol Oncol Res 2019; 26:2049-2056. [PMID: 31444708 DOI: 10.1007/s12253-019-00727-9] [Citation(s) in RCA: 18] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 06/25/2019] [Accepted: 08/14/2019] [Indexed: 12/15/2022]
Abstract
One-step nucleic acid amplification (OSNA) is used to intraoperatively detect sentinel lymph node metastases in breast cancer. OSNA has also been proposed in endometrial cancer, but evidence in this regard is unclear to define the diagnostic accuracy of OSNA in detecting lymph node metastases in endometrial cancer. A systematic review and meta-analysis was performed by searching 8 electronic databases from their inception to March 2019 for studies testing the diagnostic accuracy of OSNA in detecting sentinel lymph node metastasis in endometrial cancer. Pathologic ultrastaging was the reference standard. Sensitivity, specificity, positive and negative likelihood ratio (LR+ and LR-), diagnostic odds ratio (DOR) and area under the curve (AUC) on SROC curve were calculated. Four studies with 237 patients and 691 lymph nodes were included. OSNA showed sensitivity = 0.88, specificity = 0.93, LR + =17.95, LR- = 0.15, DOR = 191.23 and high diagnostic accuracy (AUC = 0.959). OSNA appears as a highly accurate tool for intraoperative assessment of sentinel lymph node in endometrial cancer.
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Togami S, Kawamura T, Fukuda M, Yanazume S, Kamio M, Kobayashi H. Quantitative RT-PCR Assay for Detecting Lymph Node Metastasis in Endometrial Cancer: A Preliminary Study. Oncology 2018; 96:179-182. [PMID: 30428472 DOI: 10.1159/000493485] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/16/2018] [Accepted: 09/03/2018] [Indexed: 11/19/2022]
Abstract
OBJECTIVE The detection accuracy of sentinel lymph node (SLN) metastasis by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) for endometrial cancer (EC) remains unclear and was assessed in this preliminary study. METHODS We studied primary cancer tissues and pelvic lymph nodes (PLN) from 105 patients with EC. qRT-PCR assay was performed to determine the copy numbers of CK19 mRNA in EC tissues, and negative and positive LN samples. Further, qRT-PCR results were compared with pathological findings. RESULTS CK19 mRNA expression was detected in 98% (104/106) of the tumors, with a median copy number of 3.0 × 105/μL. Twelve LN were diagnosed as positive by pathological examination. The median copy number of CK19 mRNA for positive and negative LN was 8.1 × 104/μL and 90.4/µL, respectively. CK19 mRNA expression was higher in pathologically positive LN than in pathologically negative LN (p < 0.01); the pathological and qRT-PCR findings showed no discrepancy. When the cutoff value was set at 4,500 copies/µL, qRT-PCR assay using CK19 mRNA exhibited high sensitivity and specificity. CONCLUSIONS Our results demonstrated that qRT-PCR assay, using CK19 mRNA, exhibits a high accuracy for detecting LN metastasis in EC and represents a useful alternative to conventional pathological diagnosis of EC.
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Affiliation(s)
- Shinichi Togami
- Department of Obstetrics and Gynecology, Faculty of Medicine, Kagoshima University, Kagoshima, Japan,
| | - Toshihiko Kawamura
- Department of Obstetrics and Gynecology, Faculty of Medicine, Kagoshima University, Kagoshima, Japan
| | - Mika Fukuda
- Department of Obstetrics and Gynecology, Faculty of Medicine, Kagoshima University, Kagoshima, Japan
| | - Shintaro Yanazume
- Department of Obstetrics and Gynecology, Faculty of Medicine, Kagoshima University, Kagoshima, Japan
| | - Masaki Kamio
- Department of Obstetrics and Gynecology, Faculty of Medicine, Kagoshima University, Kagoshima, Japan
| | - Hiroaki Kobayashi
- Department of Obstetrics and Gynecology, Faculty of Medicine, Kagoshima University, Kagoshima, Japan
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Nagai T, Niikura H, Okamoto S, Nakabayashi K, Matoda M, Utsunomiya H, Nagase S, Watanabe M, Takeshima N, Yaegashi N. A new diagnostic method for rapid detection of lymph node metastases using a one-step nucleic acid amplification (OSNA) assay in endometrial cancer. Ann Surg Oncol 2014; 22:980-6. [PMID: 25190122 DOI: 10.1245/s10434-014-4038-2] [Citation(s) in RCA: 36] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/22/2014] [Indexed: 12/11/2022]
Abstract
BACKGROUND To improve lymph node (LN) metastasis identification for patients with endometrial cancer (EC), this study assessed the usefulness of molecular biologic techniques using a one-step nucleic acid amplification (OSNA) assay. METHODS Using quantitative reverse transcription polymerase chain reaction (qRT-PCR), an optimal mRNA marker was selected, and its expression was compared between histopathologically positive and negative LNs using an OSNA assay. The authors determined copy number cutoff values and evaluated the diagnostic performance of this OSNA assay using sentinel lymph nodes (SLNs). They also investigated whether an OSNA assay could detect LN metastases with sensitivity and specificity equivalent to the 2-mm-interval histopathology method. RESULTS For analysis of EC samples, cytokeratin 19 (CK19) was selected as a useful mRNA marker for the OSNA assay. When the cutoff value was set at 250 copies (using 215 LNs from 70 patients), an OSNA assay using CK19 mRNA had a sensitivity of 93.3%, a specificity of 99.5%, and a concordance rate of 99.1%. For performance evaluations using SLNs (120 histopathologically negative LNs and 17 histopathologically positive LNs from 35 patients), a OSNA assay using CK19 mRNA had a sensitivity of 82.4%, a specificity of 99.2%, a positive predictive value of 93.3%, and a concordance rate of 97.1%. Thus, an OSNA assay using CK19 mRNA provided results equivalent to those with the 2-mm-interval histopathology method. CONCLUSIONS The study data demonstrated that an OSNA assay using CK19 mRNA was applicable for detecting LN metastases in EC. Combined analysis using an OSNA assay and SLNs may improve individualized treatments according to LN metastatic status.
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Affiliation(s)
- Tomoyuki Nagai
- Department of Obstetrics and Gynecology, Tohoku University School of Medicine, Sendai, Japan
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Pathologic ultrastaging improves micrometastasis detection in sentinel lymph nodes during endometrial cancer staging. Int J Gynecol Cancer 2014; 23:964-70. [PMID: 23694985 DOI: 10.1097/igc.0b013e3182954da8] [Citation(s) in RCA: 205] [Impact Index Per Article: 18.6] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/18/2022] Open
Abstract
OBJECTIVE To describe the incidence of low-volume ultrastage-detected metastases in sentinel lymph nodes (SLNs) identified at surgical staging for endometrial carcinoma and to correlate it with depth of myoinvasion and tumor grade. METHODS We reviewed all patients who underwent primary surgery for endometrial carcinoma with successful mapping of at least one SLN at our institution from September 2005 to December 2011. All patients underwent a cervical injection for mapping. The SLN ultrastaging protocol involved cutting an additional 2 adjacent 5-μm sections at each of 2 levels, 50-μm apart, from each paraffin block lacking metastatic carcinoma on routine hematoxylin and eosin (H&E) staining. At each level, one slide was stained with H&E and with immunohistochemistry (IHC) using anticytokeratin AE1:AE3.Micrometastases (tumor deposits >0.2 mm and ≤2 mm) and isolated tumor cells (≤0.2 mm) were classified as low-volume ultrastage-detected metastases if pathologic ultrastaging was the only method allowing detection of such nodal disease. RESULTS Of 508 patients with successful mapping, 413 patients (81.3%) had endometrioid carcinoma. Sixty-four (12.6%) of the 508 patients had positive nodes: routine H&E detected 35 patients (6.9%), ultrastaging detected an additional 23 patients (4.5%) who would have otherwise been missed (4 micrometastases and 19 isolated tumor cells), and 6 patients (1.2%) had metastatic disease in their non-SLNs. The incidence rates of low-volume ultrastage-detected nodal metastases in patients with grades 1, 2, and 3 tumors were 3.8%, 3.4%, and 6.9%, respectively. The frequency rates of low-volume ultrastage-detected metastases in patients with a depth of myoinvasion of 0, less than 50%, and 50% or more were 0.8%, 8.0%, and 7.4%, respectively. Lymphovascular invasion was present in 20 (87%) of the cases containing low-volume ultrastage-detected metastases in the lymph nodes. CONCLUSIONS Sentinel lymph node mapping with pathologic ultrastaging in endometrial carcinoma detects additional low-volume metastases (4.5%) that would otherwise go undetected with routine evaluations. Our data support the incorporation of pathologic ultrastaging of SLNs in endometrial carcinoma with any degree of myoinvasion. The oncologic significance of low-volume nodal metastases requires long-term follow-up.
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Comparative assessment of lymph node micrometastasis in cervical, endometrial and vulvar cancer: insights on the real time qRT-PCR approach versus immunohistochemistry, employing dual molecular markers. BIOMED RESEARCH INTERNATIONAL 2014; 2014:187684. [PMID: 24527437 PMCID: PMC3910066 DOI: 10.1155/2014/187684] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 08/02/2013] [Accepted: 10/23/2013] [Indexed: 02/06/2023]
Abstract
To address the value of qRT-PCR and IHC in accurately detecting lymph node micrometastasis in gynecological cancer, we performed a systematic approach, using a set of dual molecular tumor-specific markers such as cytokeratin 19 (CK19) and carbonic anhydrase 9 (CA9), in a series of 46 patients (19 with cervical cancer, 18 with endometrial cancer, and 9 with vulvar cancer). A total of 1281 lymph nodes were analyzed and 28 were found positive by histopathology. Following this documentation, 82 lymph nodes, 11 positive and 71 negative, were randomly selected and further analyzed both by IHC and qRT-PCR for CK19 and CA9 expression. All 11 (100%) expressed CK19 by IHC, while only 6 (54.5%) expressed CA9. On the contrary, all the histologically negative for micrometastases lymph nodes were also negative by IHC analysis for both markers. The comparative diagnostic efficacy of the two markers using qRT-PCR, however, disclosed that the analysis of the same aliquots of the 82 lymph nodes led to 100% specificity for the CK19 biomarker, while, in contrast, CA9 failed to recapitulate a similar pattern. These data suggest that qRT-PCR exhibits a better diagnostic accuracy compared to IHC, while CK19 displays a consistent pattern of detection compared to CA9.
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Han G, Lim D, Leitao MM, Abu-Rustum NR, Soslow RA. Histological features associated with occult lymph node metastasis in FIGO clinical stage I, grade I endometrioid carcinoma. Histopathology 2013; 64:389-98. [PMID: 24215212 DOI: 10.1111/his.12254] [Citation(s) in RCA: 38] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/18/2013] [Accepted: 08/09/2013] [Indexed: 11/26/2022]
Abstract
AIMS Lymph node involvement affects prognosis/treatment in endometrial carcinoma patients. We assessed various histological features associated with nodal metastasis in patients with grade I, stage I endometrial endometrioid carcinoma (EEC). METHODS AND RESULTS Eighteen stage I EECs with occult positive lymph nodes and 36 controls were assessed for depth of myoinvasion; microcystic, elongated and fragmented (MELF) pattern of myometrial invasion; lymphovascular invasion (LVI); and epithelial metaplasia. Nodal metastases were subclassified as isolated tumour cells (ITCs; ≤0.2 mm), micrometastasis (>0.2 mm and <2 mm), or macrometastasis (≥2 mm). Node-positive cases had significantly higher rates of LVI (P < 0.001) and MELF invasion (P = 0.003) on univariate analysis. Only LVI was associated significantly with nodal metastasis on multivariate analysis (P = 0.002). Tumours with MELF invasion demonstrated reduced E-cadherin expression. Macrometastases were identified in seven cases (39%) with or without micrometastasis/ITCs. Eight (44%) contained only ITCs. Eleven (61%) had histiocyte-like nodal metastases. Biopsy material from four of six (67%) and five of 17 (29%) cases with and without nodal metastasis showed detached eosinophilic tumour cell buds. Of the former, three were associated with histiocyte-like nodal metastases - a feature absent in biopsies without tumour budding. CONCLUSIONS Lymph nodes from grade I EEC exhibiting cellular budding or LVI should be examined for occult metastases, especially in the form of histiocyte-like cells.
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Affiliation(s)
- Guangming Han
- Department of Pathology, Sunnybrook Health Sciences Centre, University of Toronto, Toronto, ON, Canada
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Turan T, Hizli D, Sarici S, Boran N, Gundogdu B, Karadag B, Tulunay G, Kose MF. Is it possible to predict para-aortic lymph node metastasis in endometrial cancer? Eur J Obstet Gynecol Reprod Biol 2011; 158:274-9. [PMID: 21664758 DOI: 10.1016/j.ejogrb.2011.04.031] [Citation(s) in RCA: 39] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/23/2010] [Revised: 04/01/2011] [Accepted: 04/30/2011] [Indexed: 10/18/2022]
Abstract
OBJECTIVE The purpose of this study was to determine the histopathologic risk factors for pelvic lymph node (PLN) and para-aortic lymph node (PALN) metastasis in endometrial cancer (EC) and to identify in which patients PALN dissection should be performed. STUDY DESIGN A total of 204 consecutive patients, with EC and underwent systematic pelvic and para-aortic lymphadenectomy extending to the renal vessels, were studied retrospectively. Statistical significance between risk factors was examined using multivariant logistic regression analysis. RESULTS Cell type, depth of myometrial invasion and tumor size were found to be independently related to PLN metastasis. PLN metastasis in any site and lymphovascular invasion (LVSI) were independent prognostic factors for predicting PALN metastasis. The sensitivity, specificity and the NPV of PLN metastasis for detecting PALN metastasis were 80.8%, 89.3% and 97%, respectively. Furthermore, the 204 patients were divided into two groups according to the presence of one of these following factors: (1) non-endometrioid cell type, (2) PLN metastasis, (3) LVSI, (4) adnexal metastasis and (5) serosal involvement. Among these 204 patients, 104 had one or more of these factors (group A), and 100 patients had none of these factors (group B). PALN metastasis was significantly greater in group A, compared to group B. The sensitivity and the NPV of these combined prognostic factors for predicting PALN metastasis were 96.2% and 99%, respectively. CONCLUSIONS Presence of non-endometrioid cell type, PLN metastasis, LVSI, adnexal metastasis or serosal involvement diagnosed by frozen section (FS) seem to be poor prognostic factor for PALN metastasis in EC. Also, PALN dissection should be extended to the level of the renal vessels in all patients who will undergo PALN dissection, due to frequent involvement of the supramesenterial region.
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Affiliation(s)
- Taner Turan
- Etlik Zübeyde Hanım Women's Health Teaching and Research Hospital, Gynecologic Oncology Division, Ankara, Turkey
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Bézu C, Coutant C, Ballester M, Feron JG, Rouzier R, Uzan S, Daraï E. Ultrastaging of lymph node in uterine cancers. JOURNAL OF EXPERIMENTAL & CLINICAL CANCER RESEARCH : CR 2010; 29:5. [PMID: 20092644 PMCID: PMC2828991 DOI: 10.1186/1756-9966-29-5] [Citation(s) in RCA: 27] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Subscribe] [Scholar Register] [Received: 09/28/2009] [Accepted: 01/21/2010] [Indexed: 11/10/2022]
Abstract
Background Lymph node status is an important prognostic factor and a criterion for adjuvant therapy in uterine cancers. While detection of micrometastases by ultrastaging techniques is correlated to prognosis in several other cancers, this remains a matter of debate for uterine cancers. The objective of this review on sentinel nodes (SN) in uterine cancers was to determine the contribution of ultrastaging to detect micrometastases. Methods Review of the English literature on SN procedure in cervical and endometrial cancers and histological techniques including hematoxylin and eosin (H&E) staining, serial sectioning, immunohistochemistry (IHC) and molecular techniques to detect micrometastases. Results In both cervical and endometrial cancers, H&E and IHC appeared insufficient to detect micrometastases. In cervical cancer, using H&E, serial sectioning and IHC, the rate of macrometastases varied between 7.1% and 36.3% with a mean value of 25.8%. The percentage of women with micrometastases ranged from 0% and 47.4% with a mean value of 28.3%. In endometrial cancer, the rate of macrometastases varied from 0% to 22%. Using H&E, serial sectioning and IHC, the rate of micrometastases varied from 0% to 15% with a mean value of 5.8%. In both cervical and endometrial cancers, data on the contribution of molecular techniques to detect micrometastases are insufficient to clarify their role in SN ultrastaging. Conclusion In uterine cancers, H&E, serial sectioning and IHC appears the best histological combined technique to detect micrometastases. Although accumulating data have proved the relation between the risk of recurrence and the presence of micrometastases, their clinical implications on indications for adjuvant therapy has to be clarified.
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Affiliation(s)
- Corinne Bézu
- Department of Gynaecology and Obstetrics, Hôpital Tenon, Assistance Publique des Hôpitaux de Paris, CancerEst, Université Pierre et Marie Curie, Paris 6, France
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Altgassen C, Müller N, Hornemann A, Kavallaris A, Hornung D, Diedrich K, Jarutat T. Immunohistochemical workup of sentinel nodes in endometrial cancer improves diagnostic accuracy. Gynecol Oncol 2009; 114:284-7. [DOI: 10.1016/j.ygyno.2009.04.020] [Citation(s) in RCA: 16] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/31/2008] [Revised: 04/15/2009] [Accepted: 04/17/2009] [Indexed: 10/20/2022]
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Delpech Y, Coutant C, Darai E, Barranger E. Sentinel lymph node evaluation in endometrial cancer and the importance of micrometastases. Surg Oncol 2008; 17:237-45. [DOI: 10.1016/j.suronc.2008.04.001] [Citation(s) in RCA: 36] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/25/2022]
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Niikura H, Okamoto S, Yoshinaga K, Nagase S, Takano T, Ito K, Yaegashi N. Detection of micrometastases in the sentinel lymph nodes of patients with endometrial cancer. Gynecol Oncol 2007; 105:683-6. [PMID: 17442382 DOI: 10.1016/j.ygyno.2007.01.052] [Citation(s) in RCA: 30] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/24/2006] [Revised: 01/24/2007] [Accepted: 01/29/2007] [Indexed: 11/27/2022]
Abstract
OBJECTIVE We investigated the utility of sentinel lymph node (SLN) mapping for the detection of endometrial carcinoma micrometastases. We reevaluated the accuracy of our SLN detection procedure, this time combining step-serial section with cytokeratin immunostaining. PATIENTS AND METHODS Between March 2002 and March 2005, consecutive patients undergoing laparotomy (total abdominal hysterectomy, bilateral salpingo-oophorectomy, total pelvic lymphadenectomy and para-aortic lymphadenectomy to the level of renal veins) with SLN biopsy for endometrial cancer at Tohoku University Hospital were enrolled in this study. Excluded were patients in whom lymph node metastases were detected by routine histological examination or those without detectable SLNs. All surgically removed lymph nodes, including SLNs, were examined histopathologically by immunohistochemistry staining with an anti-cytokeratin antibody (AE1/AE3) combined with step-serial sectioning at 200-500 microm intervals. RESULTS Four of seventy-four SLNs (5%) obtained from 20 patients had micrometastases or isolated tumor cells (ITC). In contrast, only 4 of the 1350 non-SLNs obtained from 20 patients (0.3%) had detectable micrometastases. The micrometastases were detected in the external iliac basin (two cases) and in the para-aortic area (two cases). The isolated tumor cell was detected in the external iliac basin (one case). CONCLUSION SLNs detected by our method had micrometastases more frequently than did non-SLNs. Easy detection of micrometastases by immunostaining is only possible with step-serial sectioning of the SLNs.
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Affiliation(s)
- Hitoshi Niikura
- Department of Obstetrics and Gynecology, Tohoku University School of Medicine, 1-1, Seiryo-machi, Sendai 980-8574, Japan.
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Tong X, Yang L, Lang JC, Zborowski M, Chalmers JJ. Application of immunomagnetic cell enrichment in combination with RT-PCR for the detection of rare circulating head and neck tumor cells in human peripheral blood. CYTOMETRY PART B-CLINICAL CYTOMETRY 2007; 72:310-23. [PMID: 17205568 DOI: 10.1002/cyto.b.20177] [Citation(s) in RCA: 50] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/30/2023]
Abstract
Detection of rare, circulating tumor cells (CTC's) in human peripheral blood is a potential indicator of prognosis and diagnosis in oncology. Typical methods to detect these CTC's are either by immunocytochemistry (ICCS) or RT-PCR. However without accurate, rapid, and reproducible enrichment processes, these detection techniques are labor intensive and/or unreliable. In this article, a repeatable enrichment process that included a flow-through immunomagnetic cell separation system, the quadrupole magnetic sorter (QMS) was optimized with the aid of a statistical analysis software package. The QMS was operated in a negative mode of operation by immunomagnetically targeting normal human peripheral blood lymphocytes (PBL) through the CD45 surface marker. Three head and neck squamous carcinoma cell lines (HNSCC), Detroit-562, SCC-4, and CAL-27, were used to determine the sensitivity of RT-PCR for the epidermal growth factor receptor (EGFR) in spiked PBL. The detection purity needed for detection was found to be one cell in 10(4), one cell in 10(3), and one cell in 10(5) for the Detroit-562, SCC-4, and CAL-27, respectively. The actual number of cancer cells needed for RT-PCR detection ranged from 30 to 1 cell. To mimic the potential concentration of rare CTC present in peripheral blood of cancer patients, the spiking concentration was chosen to be one cancer cell per 10(5) total leukocytes from healthy donors. Using a single step immunomagnetic labeling, the final, optimized enrichment process produced a 57.6 +/- 30.3-fold (n = 6) enrichment of the rare cancer cells with a final cancer cell recovery of (77.8 +/- 6.6)%.
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Affiliation(s)
- Xiaodong Tong
- Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio 43210, USA
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Ji XQ, Sato H, Tanaka H, Konishi Y, Fujimoto T, Takahashi O, Tanaka T. Real-time quantitative RT-PCR detection of disseminated endometrial tumor cells in peripheral blood and lymph nodes using the LightCycler System. Gynecol Oncol 2005; 100:355-60. [PMID: 16203027 DOI: 10.1016/j.ygyno.2005.08.041] [Citation(s) in RCA: 12] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/16/2005] [Revised: 08/01/2005] [Accepted: 08/30/2005] [Indexed: 10/25/2022]
Abstract
OBJECTIVE Some endometrial cancer patients without clinical evidence of extrauterine spread die as a result of recurrence even after curative operation. These recurrences may arise from occult tumor cells that are not detected by conventional methods. The goal of this study was to develop a quantitative method for the detection of disseminated tumor cells (DTCs) in the peripheral blood (PB) and lymph nodes (LNs) of patients with endometrial cancer. METHODS Ninety-eight PB samples from 30 patients and 218 LNs from 14 patients were studied. Real-time quantitative analysis was performed using a LightCycler instrument and a TaqMan probe for cytokeratin 19 (CK19) as a marker gene. RESULTS This method resulted in the reproducible quantitation of 10 to 10(6) MCF-7 cells (CK19-expressing breast cancer cell line) per 10(6) peripheral blood nucleated cells. CK19 mRNA expression was detected in 28 PB samples and in 62 LNs. Only three preoperative PB samples and one postoperative PB sample (from four patients) and 33 LNs (from six patients) were PCR-positive. The PCR-positive rate of LNs was higher in patients with pathologically metastatic (path-positive) LNs than in patients with path-negative but PCR-positive LNs. Furthermore, the CK19 mRNA background expression rate was higher in the LNs of path-negative but PCR-positive patients than in LNs of path-negative and PCR-negative patients. CONCLUSIONS Real-time qRT-PCR with TaqMan probes is a sensitive, specific and rapid method for the detection of DTCs in PB and LNs. Additional studies with larger numbers of patients and adequate follow-up would be of benefit.
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Affiliation(s)
- Xin-Qiang Ji
- Akita University School of Medicine, Division of Obstetrics and Gynecology, Department of Reproductive and Developmental Medicine, 1-1-1 Hondo, Akita-shi, Akita 010-8543, Japan
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Horn LC, Einenkel J, Höckel M, Kölbl H, Kommoss F, Lax SF, Riethdorf L, Schnürch HG, Schmidt D. Pathologisch-anatomische Aufarbeitung und Befundung von Lymphknoten bei gynäkologischen Malignomen. DER PATHOLOGE 2005; 26:266-72. [PMID: 15915329 DOI: 10.1007/s00292-005-0764-3] [Citation(s) in RCA: 27] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/28/2022]
Abstract
The nodal status is one of the strongest prognostic factors in gynecologic malignancies. Metastatic involvement of regional and distant lymph nodes represents the selection basis for adjuvant therapy in a large number of solid neoplasms. The number of resected lymph nodes is one of the most important parameters in the quality control of the surgical procedure, in particular with respect to radicality. The present paper provides recommendations for gross dissection, laboratory procedures and reporting for lymph node biopsies, lymph node dissections and sentinel lymph node biopsies (SLN) for cancers of the vulva, vagina, uterine cervix, endometrium, Fallopian tubes and the ovaries, submitted for the evaluation of metastatic disease. The pathologic oncology report should include information about the number and size of resected lymph nodes, the number of involved lymph nodes with the maximum size of metastases and the presence of paranodal infiltration. In addition, the detection of isolated tumor cells should be reported, particularly with respect to the detection method (immunostains or molecular methods). In cases of metastatic disease and carcinoma of unknown primary (CUP-syndrome), information should be given regarding the primary tumor.
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Affiliation(s)
- L-C Horn
- Abteilung für Gynäko- & Perinatalpathologie, Institut für Pathologie der Universität Leipzig.
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Lara O, Tong X, Zborowski M, Chalmers JJ. Enrichment of rare cancer cells through depletion of normal cells using density and flow-through, immunomagnetic cell separation. Exp Hematol 2004; 32:891-904. [PMID: 15504544 DOI: 10.1016/j.exphem.2004.07.007] [Citation(s) in RCA: 141] [Impact Index Per Article: 6.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/06/2003] [Revised: 07/06/2004] [Accepted: 07/12/2004] [Indexed: 10/26/2022]
Abstract
OBJECTIVE To develop a reliable technique to enrich for rare cells in blood suspensions using only negative selection steps including a flow-through immunomagnetic cell separations system and by optimizing variables normally encountered during such enrichment processes. METHODS A human breast cancer cell line was cultivated and spiked at a ratio of 1 cancer cell to 10(5) total leukocytes in buffy coat or 1 cancer cell to 10(8) total cells in whole blood samples. The final, optimized process consisted of: a red cell lysis step, immunomagnetically staining leukocytes with an anti-CD45 PE, anti- MACS sandwich, immunomagnetic sorting using a flow-through system (QMS), and a final cell analysis step using either an automated cell counter, filtration, and visual counting or a cytospin analysis. RESULTS The final, optimized process produced a final enrichment of the rare cancer cells of 5.17 log(10) and an average, final recovery of 46%. It should be noted that a negative depletion protocol was used (i.e., no labeling of the rare cancer cells was used). CONCLUSIONS To the authors' knowledge, no examples in the literature exist of a 5.17 log(10) enrichment of cancer cells in human blood using a negative depletion protocol. The closest example is a 4 log(10) enrichment in which two positive magnetic cell separation steps were used (none were used in this study). Ongoing studies are investigating further modifications of the precommercial, prototype flow-through immunmagnetic separation system to increase both the enrichment and recovery rate. However, even at current performance levels, the presented process could significantly improve visual and molecular analysis of rare cells in blood.
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Affiliation(s)
- Oscar Lara
- Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio, USA
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Mariani A, Keeney GL, Aletti G, Webb MJ, Haddock MG, Podratz KC. Endometrial carcinoma: paraaortic dissemination. Gynecol Oncol 2004; 92:833-8. [PMID: 14984949 DOI: 10.1016/j.ygyno.2003.11.032] [Citation(s) in RCA: 82] [Impact Index Per Article: 3.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/11/2003] [Indexed: 01/02/2023]
Abstract
OBJECTIVE The objective of our study was to identify pathologic factors predictive of tumor dissemination to paraaortic lymph nodes (LNs) in endometrial carcinoma. The identification of the risk factors may potentially facilitate selection of patients for radical surgery or radiotherapy directed to the paraaortic area (PAA). METHODS The study population was a cohort from 612 consecutive patients with endometrial cancer surgically managed at our institution over a 10-year period. Tumor dissemination to the PAA was identified by selecting those patients who had either paraaortic LNs positive for disease at the time of primary surgery or those who subsequently experienced paraaortic failure or both (n=41; the "PA mets" subgroup). Therefore, patients for whom no information was available about the status of paraaortic LNs but who had received adjuvant irradiation to the PAA and those for whom information was not available about sites of recurrent disease were excluded from the analysis, leaving 566 patients to compose the study population. RESULTS On the basis of univariate analysis, numerous pathologic variables were significantly (P< or =0.01) associated with PA mets. However, logistic regression analysis identified only two independent factors predictive of PA mets: positive pelvic LNs (P<0.001, OR=5.00) and lymphovascular invasion (LVI) (P=0.01, OR=1.99). Notably, only 2% of patients with negative pelvic LNs had PA mets compared with 47% of those with positive pelvic LNs (P<0.001). When both pelvic LNs and LVI were negative, only 0.8% of the patients had PA mets compared with 31% of patients for whom at least one of the two variables was positive (P<0.001). CONCLUSION Positive pelvic LNs and LVI identify a subgroup of high-risk patients (approximately one sixth of the overall population) who potentially may benefit from formal lymphadenectomy or adjuvant therapy or both directed to the PAA. Furthermore, with 47% of patients with positive pelvic LNs having PA mets, unstaged patients at risk for pelvic LN involvement should be considered candidates for both pelvic and paraaortic external beam radiotherapy or surgical restaging.
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Affiliation(s)
- Andrea Mariani
- Section of Gynecologic Surgery, Mayo Clinic, Rochester, MN 55905, USA
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Querleu D, Leblanc E, Martel P, Ferron G, Narducci F. [Lymph node dissection in the surgical management of stage I endometrial carcinomas]. ACTA ACUST UNITED AC 2004; 31:1004-12. [PMID: 14680780 DOI: 10.1016/j.gyobfe.2003.06.003] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/30/2022]
Abstract
The indication and extent of lymph node dissection in the surgical management of endometrial cancers remain highly controversial. Randomized studies are badly needed but will probably lack for the next years, considering the large sample size required to show a small difference in survival. The trend towards a reduction in the routine use of external radiation therapy weakens the argument that radiation therapy makes adequate lymph node dissection useless. The balance stays between the risk for node involvement and the expected complications rate of the procedure. Lymph node dissection is advised whenever there is a non-negligible risk of node metastasis in a patient at low surgical risk.
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Affiliation(s)
- D Querleu
- Département de chirurgie, institut Claudius-Regaud, 20, rue du Pont-Saint-Pierre, 31000, Toulouse, France.
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Kamiya M, Ichiki Y, Kamiya H, Yamamoto A, Kitajima Y. Detection of nonmelanoma skin cancer micrometastases in lymph nodes by using reverse transcriptase-polymerase chain reaction for keratin 19 mRNA. Br J Dermatol 2003; 149:998-1005. [PMID: 14632805 DOI: 10.1111/j.1365-2133.2003.05602.x] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/28/2022]
Abstract
BACKGROUND A new sensitive method for the detection of skin cancer micrometastases in lymph nodes is based on amplification of keratin 19 (K19) mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR). OBJECTIVES To compare results of RT-PCR with those of histological examination in terms of the detection rate of skin cancer micrometastases. METHODS Twenty-six lymph nodes obtained from 13 patients with squamous cell carcinoma (SCC), eccrine porocarcinoma and Paget's disease were investigated by histological examination (haematoxylin and eosin sections) and RT-PCR. RT-PCR was performed on extracted RNA by using K19 primer pairs. RT-PCR products were visualized by ethidium bromide staining and confirmed by non-radioactive hybridization with K19-specific probes. RESULTS All of 10 histologically positive lymph nodes yielded the expected 460-bp band. Of the 16 histologically negative lymph nodes, one (6%) was found by RT-PCR to express K19 mRNA, indicating the presence of micrometastases which could not be detected by histological examination. A serial dilution study using RNA extracted from SCC cells mixed with RNA extracted from normal lymph node cells showed a detection sensitivity of K19 RT-PCR of 10-5 micro g cancer cell RNA in 1 micro g lymph node RNA. Nested RT-PCR showed a detection sensitivity of one tumour cell in 106 lymphocytes. CONCLUSIONS These results demonstrate the usefulness of K19 RT-PCR for the detection of skin cancer micrometastases in lymph nodes.
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Affiliation(s)
- M Kamiya
- Department of Dermatology, Gifu University School of Medicine, Tsukasamachi 40, Gifu 500-8705, Japan
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Alexander-Sefre F, Salvesen HB, Ryan A, Singh N, Akslen LA, MacDonald N, Wilbanks G, Jacobs IJ. Molecular assessment of depth of myometrial invasion in stage I endometrial cancer: a model based on K-ras mutation analysis. Gynecol Oncol 2003; 91:218-25. [PMID: 14529685 DOI: 10.1016/s0090-8258(03)00505-5] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
Abstract
INTRODUCTION Overall nearly 20% of endometrial cancer (EC) patients die of the disease and over half of these had initially presented with clinical stage I disease. There is a strong correlation between disease mortality and depth of myometrial invasion. Current assessment of depth of invasion relies on light microscopy. Tumor cells can evade detection by light microscopy if they are vastly outnumbered by myometrial cells. Molecular techniques have a great potential in the detection of such isolated cells. OBJECTIVE The objective was to develop a model for the application of molecular techniques to advance the assessment of risk status in patients with clinical stage I EC. METHODS The study sample included 21 stage I ECs with a documented K-ras mutation from two series of 96 and 106 ECs from the United Kingdom and Norway, respectively. K-ras was documented using heteroduplex mobility analysis and amplified created restriction site, followed by sequencing to identify the specific base substitution at codon 12 and 13 of K-ras oncogene. For each case with a K-ras mutation, a modified mutant allele-specific amplification technique was carried out on a series of tissue strips microdissected at increasing depths from the myometrium underlying tumor. The microdissected myometrium had been previously examined histologically for absence of infiltrating tumor cells on light microscopy. Presence of K-ras mutations was used to identify the tumor cells within the histologically normal myometrium. Correlations between submicroscopic myometrial tumor cell infiltration and clinicopathological factors were studied. RESULTS Of 21 cases with K-ras mutation, 6 cases (28%) showed molecular evidence of tumor cell infiltration beyond the histological boundary. The depth of submicroscopic myometrial infiltration was found to be variable. The staging of the tumors would have changed in 3 cases (14%) if tumor cells been detected histologically. A borderline significant correlation between presence of submicroscopic myometrial invasion and depth of myometrial invasion was noted (P = 0.053). The recurrence rate and survival of patients without submicroscopic invasion were better than those with, although it did not reach statistical significance (recurrence rate P = 0.13, recurrence free survival P = 0.14, cause-specific survival P = 0.12, and total survival P = 0.2). CONCLUSIONS Molecular assessment of depth of myometrial invasion using K-ras mutation is feasible and may add information to conventional light microscopy. Further prospective studies are required to define the clinical significance of this technology.
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Affiliation(s)
- Farhad Alexander-Sefre
- ICRF Translational Oncology Laboratory, St. Bartholomew's and the Royal London Medical and Dental School, Charterhouse Square, London, EC1M 6BQ, UK.
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Roggel F, Hocke S, Lindemann K, Sinz S, Welk A, Bosl M, Pabst M, Nusser N, Braun S, Schmitt M, Harbeck N. Minimal residual disease in breast cancer and gynecological malignancies: phenotype and clinical relevance. Recent Results Cancer Res 2003; 162:89-100. [PMID: 12790324 DOI: 10.1007/978-3-642-59349-9_8] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/06/2023]
Abstract
In breast cancer, about 35% of patients without any clinical signs of overt distant metastases already have disseminated tumor cells in bone marrow aspirates at the time of primary therapy. A significant prognostic impact of these disseminated tumor cells has been shown by many international studies: patients with tumor cells in their bone marrow have a significantly worse prognosis than those without them. Even in malignancies where the skeletal system is not a preferred location for distant metastasis, such as ovarian cancer, early presence of minimal residual disease (MRD) is correlated with poor patient outcome. Thus, besides analysis of the primary tumor, detection of MRD can be used for assessment of patient prognosis and for prediction or monitoring of response to systemic therapy. Disseminated tumor cells are also the targets for novel tumor biological therapy approaches such as specific antibody-based therapies against target cell-surface antigens such as HER2, Ep-CAM (17-1A), and uPA-R. In breast cancer, a first antibody-based tumor therapy against HER2 (Herceptin) has already been approved for clinical use in recurrent disease. However, patient selection for such tumor biological therapies becomes rather difficult due to phenotype changes, which may manifest themselves as differences between primary lesion and disseminated tumor cells. Therefore, not only identification of disseminated tumor cells but even more so their characterization at the protein and gene levels have become increasingly important. In conclusion, characterization of tumor biological properties of disseminated tumor cells allows identification of patients with breast cancer or gynecological malignancies at risk for relapse who are likely to benefit from systemic treatment and/or novel tumor biological therapy approaches.
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Affiliation(s)
- Frigga Roggel
- Klinische Forschergruppe der Frauenklinik, Technische Universität München, Ismaninger Strasse 22, 81675 Munich, Germany
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Weber T, Lacroix J, Wörner S, Weckauf H, Winkler S, Hinz U, Schilling T, Frank-Raue K, Klar E, Knebel Doeberitz Mv MV. Detection of hematogenic and lymphogenic tumor cell dissemination in patients with medullary thyroid carcinoma by cytokeratin 20 and preprogastrin-releasing peptide RT-PCR. Int J Cancer 2003; 103:126-31. [PMID: 12455065 DOI: 10.1002/ijc.10804] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/29/2022]
Abstract
Despite an extensive surgical approach only 50% of the patients with medullary thyroid carcinoma (MTC) are biochemically cured. The failure to cure a larger number of patients is a result of the early dissemination of MTC. The present study evaluates two RT-PCR based assays for the detection of disseminated tumor cells in blood, bone marrow and lymph node samples of patients with MTC. Frozen tissue and blood samples of 19 patients with MTC and 61 cervical lymph nodes of these patients were obtained intraoperatively during thyroidectomy and lymphadenectomy. Preoperative bone marrow samples were obtained from 8 patients with MTC. An expression of CK20 and preproGRP was found in all MTC tissue samples. Using CK20-PCR, disseminated MTC cells were detected in 67% of the cervical lymph nodes of patients with MTC, compared to 72% involved lymph nodes, detected by preproGRP-PCR. In 16 of 61 nodes (26%) each PCR-system detected disseminated tumor cells in histologically tumor-free lymph nodes. Disseminated tumor cells were detected with CK20-PCR and preproGRP in 5 of 18 (28%) preoperative blood samples, each. The detection of a hematogenic tumor cell dissemination by preproGRP correlated significantly with the tumor stages (p = 0.019). Circulating MTC cells were found in 3 of 8 bone marrow samples with CK20-PCR, compared to 1 of 8 samples with preproGRP-PCR. Both PCR assays are highly sensitive to detect disseminated MTC cells in blood, bone marrow and lymph node samples. Our results of disseminated MTC cells in 26% of histologically tumor-free cervical lymph nodes and in 28% of the blood samples of patients with MTC might therefore explain the low biochemical cure rates.
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Affiliation(s)
- Theresia Weber
- Department of Surgery, University of Heidelberg, Germany.
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Stackievicz R, Drucker L, Zemer R, Klein A, Markovitch O, Yarkoni S. Cytokeratin 20 as a biomarker of gestational trophoblastic disease: diagnostic and prognostic significance. Gynecol Oncol 2002; 87:34-8. [PMID: 12468339 DOI: 10.1006/gyno.2002.6799] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
Abstract
OBJECTIVES This study was designed to examine whether cytokeratin 20 (CK20) is expressed in molar pregnancies and may therefore be used in the diagnosis of gestational trophoblastic disease (GTD). The potential of CK20 expression in predicting the evolution and the prognosis of the different subtypes of GTD was also assessed. METHODS A total of 48 samples were studied for CK20 expression by RT-PCR methodology. Among these, 24 samples were obtained by curettage of the uterine cavity of patients diagnosed with hydatidiform mole (14 complete moles and 10 partial moles), 4 samples were obtained from choriocarcinoma cell lines (2 JAR and 2 JEG), and 20 samples were of normal trophoblast (control group) obtained from patients that underwent elective termination of pregnancy. RESULTS Expression of CK20 was identified in all the samples of complete mole (CM), all choriocarcinoma cell lines, and 50% of the patients with partial mole (PM). None of the preparations of normal trophoblastic tissue from the control group expressed the CK20. A significant difference (P < 0.00001) was found in CK20 expression between samples of patients with GTD and control samples. Comparison between CK20 expression in CMs and PMs revealed a significantly more frequent expression of CK20 in CMs (P = 0.006). More than 50% of the patients with PMs that were positive for CK20 had an invasive evolution. CONCLUSIONS In our opinion, CK20 may assist in distinguishing between molar and normal trophoblastic tissue and may be considered a marker of GTD. In cases in which pathological classification of different subtypes of GTD is in doubt, CK20-positive expression is suggestive for a CM whereas CK20-negative is more indicative for PM.
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Affiliation(s)
- Rodica Stackievicz
- Oncogenetics Laboratory, Sapir Medical Center, Meir Hospital, Kfar Sava and Sackler School of Medicine, Tel-Aviv 44281, Israel.
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Altaras MM, Klein A, Zemer R, Zimlichman S, Bernheim J, Fishman A. Detection of tumor circulating cells by cytokeratin-20 in the blood of patients with granulosa cell tumors. Gynecol Oncol 2002; 86:330-6. [PMID: 12217756 DOI: 10.1006/gyno.2002.6766] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
Abstract
OBJECTIVE Cytokeratins (CKs) are constituents of the intermediate filaments of epithelial cells which are expressed in various combinations, depending on the epithelial type and the degree of differentiation. Using the reverse transcriptase-polymerase chain reaction (RT-PCR) technique, we recently demonstrated that cytokeratin-20 (CK-20), the most recently discovered cytokeratin, is expressed in endometrial carcinoma tumors, in blood, and in lymph nodes with micrometastases of patients treated for endometrial carcinomas. However, CK-20 expression could not be demonstrated in the endometrium of patients with benign diseases, in peripheral blood, in lymph nodes of healthy subjects, or in normal blood cells. The aim of this study was to examine whether CK-20 expression in blood can be used as a biomarker for the detection of the dissemination of malignant cells in patients treated for granulosa cell tumors (GCTs). METHODS In this study, we used RT-PCR to determine the expression of CK-20 in the following groups: (i) blood of patients (n = 14) treated for GCTs, (ii) GCT samples (n = 4); (iii) lymph nodes (n = 2) of patients treated for GCTs; (iv) blood from subjects with benign sex-cord-stromal tumors (n = 2); (v) normal ovaries of two menstruating women (n = 4); (vi) tumor specimens of epithelial ovarian carcinomas (EOCs) (n = 14); and (vii) blood samples (n = 18) and lymph nodes (n = 11) of healthy women. RESULTS In Group I, CK-20 was positive in the blood in 86% (12/14) of the patients. In Group II, CK-20 was positive in 100% (4/4) of the GCT samples. In Group III, CK-20 was positive in 100% (2/2) of the lymph nodes examined. In Groups IV and V, CK-20 was negative in 100% (2/2) of the blood samples and in the normal ovarian specimens (4/4) that were examined. In Group VI, CK-20 was positive in 14% (2/14) of nonmucinous EOCs. In Group VII, CK-20 was negative in 100% (18/18) of blood and in (11/11) lymph node specimens (specificity 100%). CONCLUSIONS These results indicate that RT-PCR of CK-20, because of its high sensitivity and specificity, is a potential biomarker for detecting metastases in blood and in micrometastases in lymph nodes of patients treated for GCTs.
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Affiliation(s)
- Marco M Altaras
- Gynecologic Oncology Unit, Department of Obstetrics and Gynecology, Sapir Medical Center, Kfar Saba 44281, Israel.
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Cui JH, Krueger U, Henne-Bruns D, Kremer B, Kalthoff H. Orthotopic transplantation model of human gastrointestinal cancer and detection of micrometastases. World J Gastroenterol 2001; 7:381-6. [PMID: 11819794 PMCID: PMC4688726 DOI: 10.3748/wjg.v7.i3.381] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM: To establish a relevant animal model of human gastrointestinal cancer, which can be used for repetitive investigations, so as to improve our understanding and management of carcinogenesis and cancer metastasis.
METHODS: Intact tissues of human colorectal and pancreatic cancers were transplanted in nude mice. The biological characteristics of the original and the corresponding transplanted tumors were investigated by HE staining, PAS staining and immunostaining. The metastases in the livers and lungs of nude mice were investigated by immunostaining with biotinylated mab KL-1 and by RT-PCR using CK20 specific primers.
RESULTS: There were totally 9 of 16 surgical specimens growing in nude mice subcutaneously and/or orthotopically (4 of 6 colorectal and 5 of 10 pancreatic cancer). Tumor cell content of the specimens and freezing of tissue specimens are important factors influencing the growth of transplanted tumor. In the group of fresh tumor tissues with greater than 50% tumor cell content, the success rate of the transplantation was 100% (3 cases of pancreatic cancer and 3 cases of colorectal cancer). The orthotopically transplanted tumors resemble the original tumor morphologically and biologically, including TAA expression such as CEA by immunohistochemistry, and CEA level in the serum of mice. Ki-67 labeling index and the expression of TAA especially K-ras, 17-1A and RA-96, are associated with the potential of tumor growth in nude mice. Micrometastases in the lungs and livers of tumor bearing mice can be detected by immunostaining with biotinylated mab KL-1 and CK20-specific RT-PCR.
CONCLUSION: An orthotopic transplantation model for human colon and pancreatic cancer in nude mice has been set up. We have also established sensitive detection methods with CK-immunohistochemistry and CK20-RT-PCR to study xenotransplanted human cancer and its metastatic cancer cells in the liver and lung of nude mice. This study may be helpful in understanding the mechanism of cancer metastasis and in developing new diagnostic methods and therapeutic strategies for metastases including micrometastases.
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Affiliation(s)
- J H Cui
- Department of General Surgery, First Affiliated Hospital, College of Medicine, Zhejiang University, 79 Qingchun Road, Hangzhou 310003, Zhejiang Province,China.
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