Establishment and characterization of a cholangiocarcinoma cell line (RMCCA-1) from a Thai patient.
OBJECTIVE
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 alpha-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.


CONCLUSIONS
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.
Rattanasinganchan
Panthip
P
Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand.
Leelawat
Kawin
K
Treepongkaruna
Sa-ard
SA
Tocharoentanaphol
Chintana
C
Subwongcharoen
Somboon
S
Suthiphongchai
Tuangporn
T
Tohtong
Rutaiwan
R
D000328:Adult; D001650:Bile Duct Neoplasms / Q000378:metabolism / Q000473:pathology* / Q000503:physiopathology; D001653:Bile Ducts, Intrahepatic / Q000378:metabolism / Q000473:pathology* / Q000503:physiopathology; D018395:CA-19-9 Antigen / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology; D018281:Cholangiocarcinoma / Q000378:metabolism / Q000473:pathology* / Q000503:physiopathology; D002869:Chromosome Aberrations; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D053539:Keratin-19 / Q000235:genetics / Q000378:metabolism; D053552:Keratin-7 / Q000235:genetics / Q000378:metabolism; D008297:Male; D009361:Neoplasm Invasiveness / Q000473:pathology / Q000503:physiopathology; D013785:Thailand
D018395:CA-19-9 Antigen; D053539:Keratin-19; D053552:Keratin-7