Inhibition of invasiveness of pancreatic carcinoma cell line PANC-1 by suppression of MMP-2 gene expression using

Abstract

Objective To investigate the inhibitory effects of RNA interference on expression of matrix metalloproteinase-2（MMP-2）gene and invasiveness of human pancreatic cancer cell line PANC-1 in vitro.Methods Small interference RNA targeting MMP-2 gene was designed and constructed to plasmid pGCsi-U6.Recombinant plasmids were transfected to pancreatic carcinoma PANC1 cells with Lipofectamine 2000.The efficiency of transfection was evaluated by flow cytometry.RQPCR was used to detect the expression of MMP-2 mRNA.The expression of MMP-2 protein was determined by ELISA.The invasiveness of PANC-1 cells was measured by transwell chamber experiment.MTT assay was used to detect the proliferation and growth of PANC-1 cells.Results Sequencing confirmed that the MMP-2 siRNA plasmid was successfully constructed.The best efficiency of transfecting recombinant plasmid was 82.1%.After transfection of the MMP-2 siRNA plasmid, the MMP-2 gene expression of PANC-1 cells was suppressed to 71.74 %（P〈0.05）,and protein expression of MMP-2 fell to 49.82%（P〈0.05）.The corresponding inhibition ratio of invasiveness was 33.0%（P〈0.05）.There was no marked difference in proliferation rate measured by MTT assay among different groups（P〉0.05）.Conclusions RNAi targeting MMP-2 can suppress invasiveness of PANC-1 cells in vitro.This suggests that MMP-2 could be a target for gene therapy of pancreatic carcinoma.RNAi is expected to open up a new prospect for tumor therapy. Key words: Pancreatic carcinoma; Matrix metalloproteinase-2; RNA interference; Inva-siveness