Original Article
Copyright ©2012 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Jun 28, 2012; 18(24): 3070-3080
Published online Jun 28, 2012. doi: 10.3748/wjg.v18.i24.3070
Function of chloride intracellular channel 1 in gastric cancer cells
Peng-Fei Ma, Jun-Qiang Chen, Zhen Wang, Jin-Lu Liu, Bo-Pei Li
Peng-Fei Ma, Jun-Qiang Chen, Zhen Wang, Jin-Lu Liu, Bo-Pei Li, Department of Gastrointestinal Surgery, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
Author contributions: Ma PF and Chen JQ contributed equally to this study; Chen JQ designed this study; Ma PF performed this research; Ma PF and Chen JQ analyzed the data; Wang Z, Liu JL and Li BP performed statistical analysis; Ma PF and Chen JQ interpreted the results; Ma PF and Wang Z drafted the paper; Chen JQ revised the paper.
Supported by The National Natural Science Foundation of China, No. 30560151; the Key Research Project of Guangxi Municipal Health Bureau, No. 200824; the Research Project of Guangxi Educational Department, No. 201012MS062 and No. 2011105981002M204; and the Natural Science Foundation of Guangxi, No. 0832113
Correspondence to: Jun-Qiang Chen, Professor, Department of Gastrointestinal Surgery, the First Affiliated Hospital of Guangxi Medical University, 6 Shuangyong Road, Nanning 530021, Guangxi Zhuang Autonomous Region, China. gxmufh@163.com
Telephone: +86-771-5351990 Fax: +86-771-5350031
Received: December 29, 2011
Revised: February 28, 2012
Accepted: April 9, 2012
Published online: June 28, 2012
Abstract

AIM: To investigate the effect of chloride intracellular channel 1 (CLIC1) on the cell proliferation, apoptosis, migration and invasion of gastric cancer cells.

METHODS: CLIC1 expression was evaluated in human gastric cancer cell lines SGC-7901 and MGC-803 by real time polymerase chain reaction (RT-PCR). Four segments of small interference RNA (siRNA) targeting CLIC1 mRNA and a no-sense control segment were designed by bioinformatics technology. CLIC1 siRNA was selected using Lipofectamine 2000 and transfected transiently into human gastric cancer SGC-7901 and MGC-803 cells. The transfected efficiency was observed under fluorescence microscope. After transfection, mRNA expression of CLIC1 was detected with RT-PCR and Western blotting was used to detect the protein expression. Proliferation was examined by methyl thiazolyl tetrazolium and apoptosis was detected with flow cytometry. Polycarbonate membrane transwell chamber and Matrigel were used for the detection of the changes of invasion and migration of the two cell lines.

RESULTS: In gastric cancer cell lines SGC-7901 and MGC-803, CLIC1 was obviously expressed and CLIC1 siRNA could effectively suppress the expression of CLIC1 protein and mRNA. Proliferation of cells transfected with CLIC1 siRNA3 was enhanced notably, and the highest proliferation rate was 23.3% (P = 0.002) in SGC-7901 and 35.55% (P = 0.001) in MGC-803 cells at 48 h. The G2/M phase proportion increased, while G0/G1 and S phase proportions decreased. The apoptotic rate of the CLIC1 siRNA3 group obviously decreased in both SGC-7901 cells (62.24%, P = 0.000) and MGC-803 cells (52.67%, P = 0.004). Down-regulation of CLIC1 led to the inhibition of invasion and migration by 54.31% (P = 0.000) and 33.62% (P = 0.001) in SGC-7901 and 40.74% (P = 0.000) and 29.26% (P = 0.002) in MGC-803. However, there was no significant difference between the mock group cells and the negative control group cells.

CONCLUSION: High CLIC1 expression can efficiently inhibit proliferation and enhance apoptosis, migration and invasion of gastric cancer cells in vitro. CLIC1 might be a promising target for the treatment of gastric cancer.

Keywords: Chloride intracellular channel 1; Gastric carcinoma; Small interference RNA; Apoptosis; Invasion; Migration