Basic Research
Copyright ©2005 Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jun 14, 2005; 11(22): 3392-3397
Published online Jun 14, 2005. doi: 10.3748/wjg.v11.i22.3392
Establishment and characterization of an opisthorchiasis-associated cholangiocarcinoma cell line (KKU-100)
Banchob Sripa, Saman Leungwattanawanit, Takayuki Nitta, Chaisiri Wongkham, Vajarabhongsa Bhudhisawasdi, Anucha Puapairoj, Chongrak Sripa, Masanao Miwa
Banchob Sripa, Saman Leungwattanawanit, Anucha Puapairoj, Chongrak Sripa, Department of Pathology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
Takayuki Nitta, Masanao Miwa, Institute of Basic Medical Science, University of Tsukuba, Ibaraki 305-8575, Tsukuba, Japan
Chaisiri Wongkham, Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
Vajarabhongsa Bhudhisawasdi, Department of Surgery, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
Banchob Sripa, Chaisiri Wongkham, Vajarabhongsa Bhudhisawasdi, Anucha Puapairoj, Liver Fluke and Cholangiocarcinoma Research Center, Khon Kaen University, Khon Kaen 40002, Thailand
Author contributions: All authors contributed equally to the work.
Supported by the Faculty of Medicine, Khon Kaen University, Thailand
Correspondence to: Dr. Banchob Sripa, Department of Pathology, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand. banchob@kku.ac.th
Telephone: +66-43-202024 Fax: +66-43-348375
Received: June 30, 2004
Revised: July 1, 2004
Accepted: August 2, 2004
Published online: June 14, 2005
Abstract

AIM: To establish and characterize a new cholangiocarcinoma cell line from a patient living in the Opisthorchis viverrini (O. viverrini) endemic area of Northeast Thailand.

METHODS: Fresh liver biopsy and bile specimens were obtained from a 65-year-old Thai woman with cholangiocarcinoma of the porta hepatis. After digestion, the cells were cultured in Ham’s F12 media. The established cell line was then characterized for growth kinetics, cell morphology, imm-unocytochemistry and cytogenetics. Tumorigenicity of the cell line was determined by heterotransplanting in nude mice.

RESULTS: The primary tumor was a poorly differentiated tubular adenocarcinoma. Examination of the bile revealed malignant cells with O. viverrini eggs. The cholangioc-arcinoma cell line KKU-100 was established 4 mo after the primary culture-population doubling time was 72 h. KKU-100 possesses compact and polygonal-shaped epithelial cells. Immunocytochemically, this cell line exhibited cytokeratin, EMA, CEA, and CA125, but not α-fetoprotein (AFP), CA19-9, desmin, c-met, or p53. Such protein expressions parallel those of the primary tumor. Cytogenetic analysis identified aneuploidy karyotypes with a modal chromosome number of 78 and marked chromosomal structural changes. Inoculation of KKU-100 cells into nude mice produced a transplantable, poorly differentiated aden-ocarcinoma, similar to the original tumor.

CONCLUSION: KKU-100 is the first egg-proven, Opisthorchis-associated cholangiocarcinoma cell line, which should prove useful for further investigations of the tumor biology of this cancer.

Keywords: Cholangiocarcinoma, Cell line, Establishment, Characterization, Opisthorchiasis