Original Article
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World J Gastroenterol. Jan 28, 2014; 20(4): 1030-1037
Published online Jan 28, 2014. doi: 10.3748/wjg.v20.i4.1030
Novel esophageal squamous cell carcinoma bone metastatic clone isolated by scintigraphy, X ray and micro PET/CT
Bi-Zeng Zhao, Jie Cao, Jin-Chen Shao, Yan-Bing Sun, Li-Min Fan, Chun-Yu Wu, Sheng Liang, Bao-Feng Guo, Guang Yang, Wen-Hui Xie, Qing-Cheng Yang, Shun-Fang Yang
Bi-Zeng Zhao, Qing-Cheng Yang, Department of Orthopedics, Shanghai Sixth Hospital of Shanghai Jiaotong University, Shanghai 200233, China
Jie Cao, Wen-Hui Xie, Shun-Fang Yang, Department of Nuclear Medicine, Shanghai Chest Hospital of Shanghai Jiaotong University, Shanghai 200030, China
Jin-Chen Shao, Department of Pathology, Shanghai Chest Hospital of Shanghai Jiaotong University, Shanghai 200030, China
Yan-Bing Sun, Department of Radiology, Shanghai Chest Hospital of Shanghai Jiaotong University, Shanghai 200030, China
Li-Min Fan, Department of Thoracic Surgery, Shanghai Chest Hospital of Shanghai Jiaotong University, Shanghai 200030, China
Chun-Yu Wu, Bao-Feng Guo, Department of Breast Surgery, Shanghai Long Hua Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
Sheng Liang, Micro PET/CT Center, Department of Nuclear Medicine, Shanghai RuiJin Hospital of Shanghai Jiaotong University, Shanghai 200025, China
Guang Yang, Kyocera Communication Systems (Shanghai) Co., Ltd, Shanghai 201203, China
Author contributions: Zhao BZ and Fan LM were responsible for animal experiments and collection of human specimens; Cao J and Xie WH contributed to the tail vein injection of radionuclide and pinhole bone scintigraphy; Yang QC participated in experimental design; Shao JC contributed to pathological diagnosis; Sun YB contributed to human conventional radiography used in the mice; Wu CY and Guo BF contributed to statistical analysis and real-time polymerase chain reaction; Liang S contributed to micro positron emission/computed tomography; Yang G contributed to images processing; Yang SF designed the study and wrote the manuscript; all authors read and approved the final version.
Supported by Shanghai Science and Technology fundamental research Grant 08140902202 and 09140901500 (to Yang SF); and National Natural Science Foundation of China Grant 30973017 (to Yang QC)
Correspondence to: Yang Shun-Fang, Associate Professor, Department of Nuclear Medicine, Shanghai Chest Hospital, Shanghai Jiaotong University, Shanghai 200030, China. yzyg@sh163.net
Telephone: +86-21-62821990 Fax: +86-21-62801109;
Received: April 24, 2013
Revised: September 21, 2013
Accepted: September 29, 2013
Published online: January 28, 2014
Processing time: 277 Days and 17.8 Hours
Abstract

AIM: To establish a Chinese esophageal squamous cell carcinoma (ESCC) cell line with high bone metastasis potency using 99mTc-methylene diphosphonate (99mTc-MDP) micro-pinhole scintigraphy, X ray and micro-positron emission tomography/computed tomography (PET/CT) for exploring the mechanism of occurrence and development in esophageal cancer.

METHODS: The cells came from a BALB/c nu/nu immunodeficient mouse, and oncogenic tumor tissue was from a surgical specimen from a 61-year-old male patient with ESCC. The cell growth curve was mapped and analysis of chromosome karyotype was performed. Approximately 1 × 106 oncogenic cells were injected into the left cardiac ventricle of immunodeficient mice. The bone metastatic lesions of tumor-bearing mice were detected by 99mTc-MDP scintigraphy, micro-PET/CT and X-ray, and were resected from the mice under deep anesthesia. The bone metastatic cells in the lesions were used for culture and for repeated intracardiac inoculation. This in vivo/in vitro experimental metastasis study was repeated for four cycles. All of the suspicious bone sites were confirmed by pathology. Real-time polymerase chain reaction was used to compare the gene expression in the parental cells and in the bone metastatic clone.

RESULTS: The surgical specimen was implanted subcutaneously in immunodeficient mice and the tumorigenesis rate was 100%. First-passage oncogenic cells were named CEK-Sq-1. The chromosome karyotype analysis of the cell line was hypotriploid. The bone metastasis rate went from 20% with the first-passage oncogenic cells via intracardiac inoculation to 90% after four cycles. The established bone metastasis clone named CEK-Sq-1BM had a high potential to metastasize in bone, including mandible, humerus, thoracic and lumbar vertebrae, scapula and femur. The bone metastasis lesions were successfully detected by micro-pinhole bone scintigraphy, micro-PET/CT, and X-ray. The sensitivity, specificity and accuracy of the micro-pinhole scintigraphy, X-ray, and micro-PET/CT imaging examinations were: 89.66%/32%/80%, 88.2%/100%/89.2%, and 88.75%/77.5%/87.5%, respectively. Some gene expression difference was found between parental and bone metastasis cells.

CONCLUSION: This newly established Chinese ESCC cell line and animal model may provide a useful tool for the study of the pathogenesis and development of esophageal carcinoma.

Keywords: Esophageal squamous cell carcinoma; Cell line; Bone metastasis; Molecular imaging; Real-time polymerase chain reaction

Core tip: We established a novel esophageal squamous cell carcinoma cell line from a surgically resected human specimen and its clone with mixed bone metastasis potency by molecular imaging including conventional radiography, bone scintigraphy, and micro-positron emission tomography/computed tomography after intracardiac inoculation of the cells into nude mice. The process was repeated in vivo and in vitro for four cycles to obtain a bone metastasis clone CEK-Sq-1BM. Some gene expression difference was compared with the primary cells and their clone by real-time reverse transcriptase polymerase chain reaction. This work may be helpful for research in bone metastases.