Establishment and characterization of an opisthorchiasis-associated cholangiocarcinoma cell line (KKU-100) - PubMed (original) (raw)

Establishment and characterization of an opisthorchiasis-associated cholangiocarcinoma cell line (KKU-100)

Banchob Sripa et al. World J Gastroenterol. 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 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.

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.

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Figures

Figure 1

Figure 1

A confluent monolayer of KKU-100 cells shows compact polygonal to spindle cells. (Phase-contrast, original magnification, ×100).

Figure 2

Figure 2

Histopathology of the primary tumor (A) and KKU-100 heterotransplanted tumors (B) in nude mice. (hematoxylin and eosin, original magnification ×200).

Figure 3

Figure 3

Immunocytochemistry of the primary tumor and KKU-100 cells. A: the primary tumor showing strong expression of cytokeratin; B: strong expression of KKU-100 cell line; C: EMA expression in both heterotransplanted tumor cells; D: EMA expression in KKU-100 cells (immunoperoxidase, original magnification ×200).

Figure 4

Figure 4

Representative G-banded karyotype of KKU-100 at passage 20 showing chromosomal abnormalities and marker chromosome (mar) : 84, +XX, +1, +2, +3, +4, +5, +8, +9, 10, +11, +12, +13, +14, +der(15)t(15;15), +16, +17, +18, +19, +20, +21, +22, +mar.

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