Original Articles
Copyright ©2009 The WJG Press and Baishideng. All rights reserved.
World J Gastroenterol. Mar 7, 2009; 15(9): 1072-1078
Published online Mar 7, 2009. doi: 10.3748/wjg.15.1072
Suppression of matrix metalloproteinase-2 via RNA interference inhibits pancreatic carcinoma cell invasiveness and adhesion
Ying-Hui Zhi, Mao-Min Song, Pi-Lin Wang, Tie Zhang, Zi-Yi Yin
Ying-Hui Zhi, Mao-Min Song, Pi-Lin Wang, Tie Zhang, Zi-yi Yin, Department of General Surgery, Affiliated Beijing Tiantan Hospital, Capital Medical University, Beijing 100050, China
Author contributions: Zhi YH performed the majority of experiments; Wang PL, Yin ZY and Zhang T provided vital reagents and analytical tools and were involved in editing the manuscript; Song MM provided financial support for this work; Zhi YH and Song MM designed the study and wrote the manuscript.
Correspondence to: Dr. Mao-Min Song, Department of General Surgery, Affiliated Beijing Tiantan Hospital, Capital Medical University, Beijing 100050, China. smaomin@sohu.com
Telephone: +86-10-67096589
Fax: +86-10-67096593
Received: October 9, 2008
Revised: January 19, 2009
Accepted: January 26, 2009
Published online: March 7, 2009
Abstract

AIM: To investigate the inhibitory effects of RNA interference (RNAi) on expression of matrix metalloproteinase-2 (MMP-2) gene and invasiveness and adhesion of human pancreatic cancer cell line, BxPC-3.

METHODS: RNAi was performed using the vector (pGPU6)-based small interference RNA (siRNA) plasmid gene silence system to specifically knock down MMP-2 expression in pancreatic cancer cell line, BxPC-3. Four groups of different specific target sequence in coding region of MMP-2 and one non-specific sequence were chosen to construct four experimental siRNA plasmids of pGPU6-1, pGPU6-2, pGPU6-3 and pGPU6-4, and one negative control siRNA plasmid of pGPU6 (-). MMP-2 expression was measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Cell proliferation and apoptosis were examined by methyl thiazolyl tetrazolium (MTT) and flow cytometry, respectively. The abilities of adhesion and invasion were detected by cell adhesion assay and cell invasion assay using Transwell chambers.

RESULTS: The expression of MMP-2 was inhibited and the inhibitory effects of different sequence varied. pGPU6-1 group had the most efficient inhibitory effect, followed by pGPU6-2 and pGPU6-3 groups. Invasiveness and adhesion were more significantly reduced in pGPU6-1, pGPU6-2 and pGPU6-3 groups as compared with pGPU6 (-) and blank control groups. However, no difference concerning cell proliferation and apoptosis was observed after transfection between experiment groups and control groups.

CONCLUSION: RNAi against MMP-2 successfully inhibited the mRNA and protein expression of MMP-2 in the pancreatic cancer cell line, BxPC-3, leading to a potent suppression of tumor cell adhesion and invasion without affecting cell proliferation and apoptosis. These findings suggest that the RNAi approach towards MMP-2 may be an effective therapeutic strategy for the clinical management of pancreatic tumor.

Keywords: Pancreatic neoplasm, Tumor metastasis, Matrix metalloproteinase-2, Small interfering RNA, Tumor invasiveness