title: Establishment and characterization of an opisthorchiasis-associated cholangiocarcinoma cell line (KKU-100).
abstract: OBJECTIVE
  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 alpha-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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Pathology, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand. banchob@kku.ac.th
authors.forename: Banchob
authors.initials: B
authors.lastname: Sripa
authors.forename: Saman
authors.initials: S
authors.lastname: Leungwattanawanit
authors.forename: Takayuki
authors.initials: T
authors.lastname: Nitta
authors.forename: Chaisiri
authors.initials: C
authors.lastname: Wongkham
authors.forename: Vajarabhongsa
authors.initials: V
authors.lastname: Bhudhisawasdi
authors.forename: Anucha
authors.initials: A
authors.lastname: Puapairoj
authors.forename: Chongrak
authors.initials: C
authors.lastname: Sripa
authors.forename: Masanao
authors.initials: M
authors.lastname: Miwa
mesh_terms: D000368:Aged; D001650:Bile Duct Neoplasms / Q000469:parasitology* / Q000473:pathology; D001653:Bile Ducts, Intrahepatic / Q000469:parasitology* / Q000473:pathology; D045744:Cell Line, Tumor; D018281:Cholangiocarcinoma / Q000469:parasitology* / Q000473:pathology; D005260:Female; D006801:Humans; D009889:Opisthorchiasis / Q000150:complications* / Q000473:pathology