siRNA沉默PAR1基因表达对大肠癌细胞侵袭力的影响

Effbct of silencing PAR1 by RNA interference on invasive potential of human colon cancer cell lines

目的:研究RNA干扰活性蛋白酶受体(Protease-activated receptors,PAR1)表达以及对人类大肠癌细胞侵袭力的影响。方法:体外化学合成PAR1基因序列特异性双链RNA(dsRNA),在脂质体(Lipid)的介导下转染入人大肠癌Lovo细胞株,荧光倒置显微镜下观察转染效率,应用RT-PCR和Western blot检测PAR1mRNA和蛋白表达,体外实验测定肿瘤细胞穿透matrigel的能力。结果:干扰片段转染组的转染Pgenesil-1-PAR1-siRNA后大肠癌Lovo细胞株的PAR1基因和蛋白的表达被PAR1-siRNA有效沉默,与阴性对照组、空白质粒转染组相比PAR1 mRNA和蛋白表达水平明显降低(P<0.05)。转染48h后测定大肠癌Lovo细胞的侵袭力明显降低(P<0.01)。结论:PAR1-siRNA能有效抑制人大肠癌细胞株PAR1基因和蛋白的表达,降低人大肠癌细胞的侵袭力。

Objective：To study the effect of RNA interference（RNAi） on PAR1 gene mRNA expression and the invasive petential of human colon cancer cell lines. Methods：Chemically synthesizeddouble stranded RNA（dsRNA） targeting PAR1 was transfected into human colon cancer cell Lovo using SiPORT Lipid. The transfection eficiency was observed under fluorescence confocal microscopy. Expression of PAR1 mRNA and protein was detected by reverse transcription polymerase chain reaction（RT-PCR） and Western blot. Cell penetrate matrigel capacity were determined by invitro experiment. Results：PARl-siRNA effectively in- hibited PAR1 mRNA and protion expression（P〈0.05）,and degraded invasion capability of Lovo cells（P〈0.01）. Conclusion： PARI-siRNA inhibits PAR1 mRNA and protein expression,degrades invasion of human colon cancer cell.