Effect of mutant p27kip1 gene on human cholangiocarcinoma cell line, QBC939

doi:10.3748/wjg.14.5344

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World Journal of Gastroenterology

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1007-9327

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Baishideng Publishing Group Inc, 7041 Koll Center Parkway, Suite 160, Pleasanton, CA 94566, USA

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World J Gastroenterol. Sep 14, 2008; 14(34): 5344-5348
Published online Sep 14, 2008. doi: 10.3748/wjg.14.5344

Effect of mutant p27^kip1 gene on human cholangiocarcinoma cell line, QBC₉₃₉

Jian Luo, Yong-Jun Chen, Wei-Yu Wang, Sheng-Quan Zou

Jian Luo, Yong-Jun Chen, Wei-Yu Wang, Sheng-Quan Zou, Department of General Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China

Author contributions: Luo J, Chen YJ, Wang WY and Zou SQ designed the research; Luo J and Chen YJ performed the research; Wang WY contributed to the new reagents/analytic tools; Wang WY analyzed the data; Luo J wrote the paper.

Supported by The National High Technology Research and Development Program of China (863 Program), No. Z2002AA214061

Correspondence to: Sheng-Quan Zou, Department of General Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China. sqzou@tjh.tjmu.edu.cn

Telephone: +86-27-83662398 Fax: +86-27-83662851

Received: December 16, 2007
Revised: August 8, 2008
Accepted: August 15, 2008
Published online: September 14, 2008

Abstract

AIM: To investigate the effects of exogenously mutated p27^kip1 (p27) on proliferation and apoptosis of human cholangiocarcinoma cell line, QBC₉₃₉in vivo.

METHODS: Adenoviral vectors were used to transfect mutated p27 cDNA into human QBC₉₃₉ cell line. Expression of p27 was detected by RT-PCR. Western blot. Cell growth, morphological change, cell cycle, apoptosis and cloning formation were determined by MTT assay and flow cytometry.

RESULTS: The expression of p27 protein and mRNA was increased significantly in QBC₉₃₉ cell line transfected with Ad-p27mt. The transfer of Ad-p27mt could significantly inhibit the growth of QBC₉₃₉ cells, decrease the cloning formation rate and induce apoptosis. p27 over expression caused cell cycle arrest at G₀/G₁ phase 72 h after infection with Ad-p27mt.

CONCLUSION: p27 may cause cell cycle arrest at G₀/G₁ phase and subsequently lead to apoptosis. Recombinant adenovirus expressing mutant p27 may be potentially useful in gene therapy for cholangiocarcinoma.

Keywords: Adenovirus; Cholangiocarcinoma; Gene therapy; Cell cycle; Apoptosis