RNAi抑制胰腺癌PANC-1细胞MMP-2 mRNA的表达

Inhibitory effect of RNA interference on expression of MMP-2 mRNA in pancreatic carcinoma cell line PANC-1

目的观察RNAi体外沉默基质金属蛋白酶2(MMP-2)基因对胰腺癌PANC-1细胞MMP-2mRNA表达的抑制作用。方法设计靶向MMP-2的siRNA,并构建到pGCsi-U6质粒,重组质粒脂质体法转染PANC-1细胞;流式细胞术检测细胞GFP表达率,RQ-PCR检测细胞MMP-2的mRNA表达水平。结果 PCR鉴定证实重组质粒构建成功,质粒转染细胞GFP表达率最高达82.1%。转染干扰质粒的PANC-1细胞MMP-2基因mRNA表达抑制了71.74%(P<0.05)。结论 RNAi明显抑制胰腺癌PANC-1细胞MMP-2mRNA的表达,可望RNAi能为胰腺癌治疗开辟新的思路。

Objective To investigate the inhibitory effects of RNA interference（RNAi） on the expression of matrix metalloproteinase-2 （MMP-2） mRNA of human pancreatic carcinoma cell line PANC-1 in vitro.Methods Small interference RNA（siRNA） targeting MMP-2 gene was designed and constructed to plasmid pGCsi-U6.Recombinant plasmids were transfected to pancreatic carcinoma cell line PANC-1 cells with lipofectamine.Green fluorescence protein（GFP） was evaluated by flow cytometry after transfection.RQ-PCR was used to detect the expression of MMP-2 mRNA.ResultsPCR confirmed that the MMP-2 siRNA plasmid was successfully constructed.The best efficiency of GFP after transfection was 82.1% in PANC-1 cells.After transfected by the MMP-2 siRNA plasmid,the MMP-2 mRNA expression of PANC-1 cells was suppressed by 71.74%（P0.05）.ConclusionRNAi targeting MMP-2 gene can obviously suppress the expression of MMP-2 mRNA of PANC-1 cells.RNAi is expected to open up a new approach to pancreatic carcinoma therapy.