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.
ABSTRACT
Objective To investigate the expression of matrix metalloproteinase-2 (MMP-2)in pancreatic carcinoma and the relationship between MMP-2 with tumour clinicopatholngical features. Methods The expression of MMP-2 was detected by S-P immunohistochemistry in 36 cases with pancreatic carcinoma and 14 normal pancreat-ic tissues. Results The positive expression rate of MMP-2 was 66.7% (24/36) in pancreatic carcinoma tissue and 14.3% (2/14) in normal pancreatic tissues (χ2 = 3. 587, P < 0.01 ) ;The expression rate of MMP-2 in pancreatic carcinoma tissue with positive-node was 86.7% ( 13/15 ) ,which was higher than that with negative-node,which was 52.4% ( 11/21 ) ( P < 0.05 ) ; As to TNM staging in pancreatic carcinoma, the expression rate of MMP-2 was 41.2% (7/17) with Ⅰ,Ⅱ staging and 89.5% (17/19) with Ⅲ,Ⅳ staging(χ2=9.418,P <0. 01 ) ;The expression rate of MMP-2 was 50.0% (5/10) ,66.7% (10/15) and 72.8% (8/11) in highly,moderately and poorly differentiated pancreatic carcinoma(P > 0.05 ). Conclusions The expression of MMP-2 is strengthened significantly in pancreatic carcinoma tissue and involved in turnout invasion and metastasis features; MMP-2 might be regarded as one more marker for the invasive properties of pancreatic carcinoma.