靶向MMP-2的RNAi抑制胰腺癌PANC-1细胞侵袭性的研究  

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

作　　者：朱学锋[1] 李德春[2] 陈益君[1] 许建伟[1] 顾继礼[1] 朱东明[2] 岑建农 

机构地区：[1]江苏省泰兴市人民医院肝胆外科,225400 [2]苏州大学附属第一医院普外科 [3]江苏省血液研究所

出　　处：《中华肝胆外科杂志》2010年第11期863-866,共4页Chinese Journal of Hepatobiliary Surgery

摘　　要：目的 观察RNAi体外沉默胰腺癌PANC-1细胞MMP-2基因对胰腺癌细胞MMP-2表达及细胞侵袭能力的抑制作用.方法 设计靶向MMP-2的siRNA并构建到pGCsi-U6质粒,重组质粒通过脂质体Lipofectamine 2000转染PANC-1细胞,流式细胞仪检测质粒的转染率,RQ-PCR检测细胞MMP-2的mRNA表达水平;ELISA检测细胞MMP-2蛋白的分泌量;Transwell小室侵袭观察各组细胞侵袭能力;MTT比色法检测各组细胞增殖活性.结果 测序鉴定证实,干扰质粒构建成功,重组质粒转染率最高达82.1%.转染干扰质粒PANC-1细胞的MMP-2基因mRNA表达抑制了71.74%（P＜0.05）;MMP-2蛋白分泌下降了49.82%（P＜0.05）;细胞侵袭能力抑制率达33.0%（P＜0.05）;细胞的增殖活性无明显变化（P＞0.05）.结论 靶向MMP-2的RNAi能抑制胰腺癌PANC-1细胞的体外侵袭能力,提示MMP-2可以作为胰腺癌基因治疗的靶点,亟待RNAi能为肿瘤治疗开创崭新局面.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.

关 键 词：胰腺癌 基质金属蛋白酶-2 RNA干扰 侵袭能力 

分 类 号：R686[医药卫生—骨科学]