Original Article
Copyright ©2011 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Jun 28, 2011; 17(24): 2924-2932
Published online Jun 28, 2011. doi: 10.3748/wjg.v17.i24.2924
Characterization of a novel rat cholangiocarcinoma cell culture model-CGCCA
Chun-Nan Yeh, Kun-Ju Lin, Tsung-Wen Chen, Ren-Ching Wu, Lee-Cheng Tsao, Ying-Tzu Chen, Wen-Hui Weng, Miin-Fu Chen
Chun-Nan Yeh, Tsung-Wen Chen, Miin-Fu Chen, Department of Surgery, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan 333, Taiwan, China
Kun-Ju Lin, Department of Nuclear Medicine; Chang Gung Memorial Hospital, Chang Gung University, Taoyuan 333, Taiwan, China
Ren-Ching Wu, Department of Pathology, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan 333, Taiwan, China
Lee-Cheng Tsao, Ying-Tzu Chen, Wen-Hui Weng, Graduate Institute of Biotechnology and Department of Chemical Engineering and Biotechnology, National Taipei University of Technology, Taipei 106, Taiwan, China
Author contributions: Yeh CN helped collect the data and wrote the manuscript; Yeh CN and Weng WH were in charge of this project and revised the manuscript; Lin KJ was in charge of the FDG-PET; Chen TW was in charge of cell-line establishment; Wu RC was in charge of IHC study; Tsao LC and Chen YT were in charge of cytogenetic study; Chen MF helped to review this paper.
Supported by Chang Gung Medical Research, Program grant 350363, National Science Council grants to Yeh CN; National Science Council grant 98-2314-B-027-001 to Weng WH
Correspondence to: Wen-Hui Weng, PhD, Graduate Institute of Biotechnology and Department of Chemical Engineering and Biotechnology, National Taipei University of Technology, Taipei 106, Taiwan, China. wwhlab@gmail.com
Telephone: +886-3-3281022 Fax: +886-3-3285818
Received: September 16, 2010
Revised: November 15, 2010
Accepted: November 22, 2010
Published online: June 28, 2011
Abstract

AIM: To characterize a culture model of rat CCA cells, which were derived from a transplantable TTA-induced CCA and designated as Chang Gung CCA (CGCCA).

METHODS: The CGCCA cells were cultured at in vitro passage 12 times on a culture dish in DMEM medium. To measure the doubling time, 103 cells were plated in a 96-well plate containing the growth medium. The cells were harvested 4 to 10 d after seeding, and a standard MTT assay was used to measure the growth. The phenotype of CACCA cell and xenograft was determined by immunohistochemical study. We also determine the chromosomal alterations of CGCCA, G-banding and spectral karyotyping studies were performed. The CGCCA cell line was transplanted into the nude mice for examining its tumorigenicity. 2-Deoxy-2-(18F)fluoro-D-glucose (FDG) autoradiography was also performed to evaluate the FDG uptake of the tumor xenograft.

RESULTS: The doubling time for the CGCCA cell line was 32 h. After transplantation into nude mice, FDG autoradiography showed that the tumors formed at the cell transplantation site had a latency period of 4-6 wk with high FDG uptake excluding necrosis tissue. Moreover, immunohistochemical staining revealed prominent cytoplasmic expression of c-erb-B2, CK19, c-Met, COX-II, EGFR, MUC4, and a negative expression of K-ras. All data confirmed the phenotypic features of the CGCCA cell line coincide with the xenograft mice tumors, indicating cells containing the tumorigenicity of CCA originated from CCA. In addition, karyotypic banding analysis showed that the diploid (2n) cell status combines with ring and giant rod marker chromosomes in these clones; either both types simultaneously appeared or only one type of marker chromosome in a pair appeared in a cell. The major materials contained in the marker chromosome were primarily identified from chromosome 4.

CONCLUSION: The current CGCCA cell line may be used as a non-K-ras effect CCA model and to obtain information and reveal novel pathways for CCA. Further applications regarding tumor markers or therapeutic targeting of CCA should be addressed accordingly.

Keywords: Cholangiocarcinoma; Rat cell line; Establishment; Characterization; Thioacetamide