Basic Research
Copyright ©2006 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Oct 28, 2006; 12(40): 6500-6506
Published online Oct 28, 2006. doi: 10.3748/wjg.v12.i40.6500
Establishment and characterization of a cholangiocarcinoma cell line (RMCCA-1) from a Thai patient
Panthip Rattanasinganchan, Kawin Leelawat, Sa-ard Treepongkaruna, Chintana Tocharoentanaphol, Somboon Subwongcharoen, Tuangporn Suthiphongchai, Rutaiwan Tohtong
Panthip Rattanasinganchan, Kawin Leelawat, Tuangporn Suthiphongchai, Rutaiwan Tohtong, Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand
Kawin Leelawat, Sa-ard Treepongkaruna, Somboon Subwongchareon, Department of Surgery, Rajavithi Hospital, Bangkok, 10400, Thailand
Kawin Leelawat, Chintana Tocharoentanaphol, Cancer Cytogenetic Unit, Chulabhorn Cancer Center, Chulabhorn Research Institute, Patumthani, 12000, Thailand
Supported by Thailand Research Fund (The Royal Golden Jubilee Ph.D. Program) and Rajavithi Hospital Fund
Correspondence to: Rutaiwan Tohtong, Assistant Professor, Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand. scrth@mahidol.ac.th
Telephone: +66-2-2015458 Fax: +66-2-3547174
Received: June 12, 2006
Revised: August 12, 2006
Accepted: September 15, 2006
Published online: October 28, 2006
Abstract

AIM: To establish and characterize a new cell line derived from peripheral cholangiocarcinoma of a Thai patient.

METHODS: The peripheral cholangiocarcinoma specimen surgically obtained from the patient was aseptically processed by washing and mincing before culturing in Ham’s F12 medium containing 10% fetal bovine serum. After 3 mo, when the cell line has become homogeneous and stabilized, several features were investigated, including growth characteristics, immunofluorescence staining for cytokeratins, expression of tumor markers, chromosomal analysis by G-banding and multicolour fluorescence in situ hybridization (mFISH), in vitro migration and invasion characteristics.

RESULTS: The RMCCA-1 cell line has been established. These cells proliferated as a monolayer with a population doubling time of 48 h. Immunofluorescence staining showed positive staining for human cytokeratin 7 and 19 verifying the biliary epithelial origin. RMCCA-1 secreted carbohydrate antigen 19-9 (CA19-9), but insignificant levels of carcinoembryonic antigen (CEA) and α-fetoprotein (AFP). Chromosome analysis identified aneuploidy karyotypes with a modal chromosome number of 59. RMCCA-1 exhibited a low level of in vitro invasiveness, but a high degree of motility. The cell line exhibited a significant number of chromosomal aberrations as shown by mFISH and G-banding methods.

CONCLUSION: A new cell line derived from peripheral cholangiocarcinoma of a Thai patient has been established. This cell line shows a low level of in vitro invasiveness, but a high degree of motility. It will serve as a valuable tool for further studies on tumor biology, molecular pathogenesis, metastatic mechanism and response to therapeutic drugs of cholangiocarcinoma.

Keywords: Cholangiocarcinoma; Cell line; Establishment; mFISH; Invasion; Migration