Analysis of metastasis suppressing function of E-cadherin in gastric cancer cells by RNAi

doi:10.3748/wjg.v11.i13.2000

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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. Apr 7, 2005; 11(13): 2000-2003
Published online Apr 7, 2005. doi: 10.3748/wjg.v11.i13.2000

Analysis of metastasis suppressing function of E-cadherin in gastric cancer cells by RNAi

Zhi-Hong Zheng, Xiu-Ju Sun, Hai-Tao Zhou, Chao Shang, Hong Ji, Kai-Lai Sun

Zhi-Hong Zheng, Xiu-Ju Sun, Hai-Tao Zhou, Chao Shang, Hong Ji, Kai-Lai Sun, Department of Genetics, China Medical University, Shenyang 110001, Liaoning Province, China

Author contributions: All authors contributed equally to the work.

Supported by the national “973” (national key program on basic research) foundation of China, No. G1998051203

Correspondence to: Kai-Lai Sun, Department of Genetics, China Medical University, North 2^nd Road, Heping District, Shenyang 110001, Liaoning Province, China. klsun@vip.sina.com

Telephone: +86-24-23256666-5325 Fax: +86-24-23265842

Received: October 29, 2004
Revised: October 30, 2004
Accepted: December 3, 2004
Published online: April 7, 2005

Abstract

AIM: To study the effect of inhibited E-cadherin expression on invasion of cancer cells.

METHODS: We designed the nucleotide sequence of siRNA corresponding to 5’ non-coding and coding sequence of E-cadherin. 21-nucleotide dssiRNA was synthesized by in vitro transcription with Ambion Silencer TM siRNA Construction Kit. siRNA was transfected into gastric cancer MKN45 using TransMessenger transfection Kit. RT-PCR and immunofluorescent assay were used to investigate the inhibition of the expression of mutated E-cadherin. Invasive ability of cancer cells was determined by Transwell assay.

RESULTS: The synthesis of E-cadherin mRNA rather than protein expression was suppressed dramatically 7 d after interference. Decreased protein expression was observed on d 10 after interference. On d 11, invasion ability was enhanced significantly.

CONCLUSION: siRNA targeted at non-coding and coding sequence of E-cadherin showed significant inhibition on mRNA and protein expression. Inhibited E-cadherin expression results in increased invasion ability of cancer cells.

Keywords: E-cadherin, RNAi, Gastric cancer