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©The Author(s) 2025. Published by Baishideng Publishing Group Inc. All rights reserved.
Detecting plasma SHOX2, HOXA9, SEPTIN9, and RASSF1A methylation and circulating cancer cells for cholangiocarcinoma clinical diagnosis and monitoring
Jing Yu, Qiu-Chen Liu, Shuang-Yan Lu, Shun Wang, Hua Zhang
Jing Yu, Department of Laboratory, Wuhan Hospital of Traditional Chinese and Western Medicine (Wuhan's TCWM Hospital), Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China
Jing Yu, Hua Zhang, Department of Laboratory, Sinopharm Dongfeng General Hospital, Hubei University of Medicine, Shiyan 442008, Hubei Province, China
Qiu-Chen Liu, Department of Laboratory, Hubei Cancer Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430079, Hubei Province, China
Shuang-Yan Lu, Department of Blood Transfusion, Wuhan Chinese and Western Medicine Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China
Shun Wang, Department of Laboratory, Wuhan Chinese and Western Medicine Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China
Co-first authors: Jing Yu and Qiu-Chen Liu.
Co-corresponding authors: Shun Wang and Hua Zhang.
Author contributions: Yu J, Zhang H and Wang S conceptualized and designed the research; Liu QC and Lu SY screened patients and acquired clinical data; Liu QC and Yu J collected blood specimen and performed laboratory analysis; Zhang H and Wang S performed Data analysis; Yu J and Liu QC wrote the paper; All the authors have read and approved the final manuscript. Yu J proposed, designed and conducted analysis, performed data analysis and prepared the first draft of the manuscript. Liu QC was responsible for patient screening, enrollment, collection of clinical data and blood specimens. Both authors have made crucial and indispensable contributions towards the completion of the project and thus qualified as the co-first authors of the paper. Both Zhang H and Wang S have played important and indispensable roles in the experimental design, data interpretation and manuscript preparation as the co-corresponding authors.
Supported by the Medical Talents of Wuhan Health and Family Planning Commission, No. 2017[51] (to Yu J); the Medical Talents of Wuhan Hospital of Traditional Chinese and Western Medicine (to Yu J); the Hubei Natural Science Foundation, No. 2023AFB1091; and Wuhan Medical Research Project, No. WX23A36 (to Yu J).
Institutional review board statement: The study was conducted in accordance with ethical guidelines and approved by the Ethics Committee of Hubei Cancer Hospital (ethical approval number: LLHBCH2023YN-002).
Informed consent statement: This study only retrospectively analyzed the data of hospitalized patients with cholangiocarcinoma, including general clinical data and plasma SHOX2, HOXA9, SEPTIN9, and RASSF1A methylation. Therefore, an approval for exemption from the subject's informed consent was agreed by the Ethics Committee of Hubei Cancer Hospital.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: All data generated or analyzed during this study are included in this published article.
Open Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
https://creativecommons.org/Licenses/by-nc/4.0/ Corresponding author: Shun Wang, Department of Laboratory, Wuhan Chinese and Western Medicine Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 215 Zhongshan Road, Wuhan 430022, Hubei Province, China.
wang_shun6688@sina.com
Received: December 16, 2024
Revised: January 17, 2025
Accepted: February 25, 2025
Published online: April 15, 2025
Processing time: 101 Days and 1.5 Hours
BACKGROUND
Cholangiocarcinoma (CCA), also known as bile duct cancer, is a devastating malignancy primarily affecting the biliary tract.
AIM
To assess their performance in clinical diagnosis and monitoring of CCA, plasma methylation and circulating tumor cells were detected.
METHODS
Plasma samples were collected from Hubei Cancer Hospital (n = 156). Plasma DNA was tested to detect SHOX2, HOXA9, SEPTIN9, and RASSF1A methylation using TaqMan PCR. Circulating tumor cells (CTCs) were detected in the peripheral blood of patients using the United States Food and Drug Administration-approved cell search system before and after clinical therapy. The CCA diagnostic value was estimated using the area under the curve. The independent prognosis risk factors for patients with CCA were estimated using Cox and logistic regression analyses.
RESULTS
The sensitivity and specificity of the four DNA plasma methylations exhibited 64.74% sensitivity and 93.88% specificity for detecting CCA. The receiver operating characteristic curve of the combined value for CCA diagnosis in plasma was 0.828 ± 0.032. RASSF1A plasma methylation was related to the prognosis of patients with CCA. We determined the prognostic hazard ratio for CCA using CTC count, tumor stage, methylation, and carbohydrate antigen 19-9 levels as key factors. Our overall survival nomogram achieved a C-index of 0.705 (0.605-0.805).
CONCLUSION
SHOX2, HOXA9, SEPTIN9, and RASSF1A plasma methylation demonstrated increased sensitivity for diagnosing CCA. RASSF1A plasma methylation and CTCs were valuable predictors to assess CCA prognosis and recurrence.
Core Tip: This study first analyzed the clinical diagnosis and monitoring value of detecting plasma SHOX2, HOXA9, SEPTIN9, and RASSF1A methylation for cholangiocarcinoma (CCA). We determined that the four DNA plasma methylations exhibited 64.58% sensitivity and 94% specificity for detecting CCA. The hazard ratio of prognosis for the risk of CCA risk was identified using the Circulating tumor cells (CTCs) count, tumor stage, methylation, and carbohydrate antigen 19-9 (CA199) levels as independent prognostic factors. We developed a predictive nomogram for CCA overall survival, age, stage, CTCs, methylation, and CA199, with a C-index of 0.705 (95%CI: 0.605-0.805). This model evaluates risk factors.
