特异性siRNA下调Cr-1基因表达对人结直肠癌细胞侵袭的影响

Inihibition of cripto-1 on invasion of human colon cancer cell

目的探讨RNA干扰(RNA interference,RNAi)沉默Cr-1(Cr-1)基因对人结直肠癌细胞侵袭的影响。方法应用Cr-1基因小干扰RNA(small interfering RNA,siRNA)转染处理结直肠癌SW480细胞后,分别采用荧光实时定量RT-PCR和蛋白质印迹检测Cr-1基因mRNA和蛋白水平,分别采用软琼脂集落培养试验和Boyden小室模型试验检测癌细胞的锚着不依赖性增殖和侵袭能力。结果 siRNA转染组Cr-1mRNA和蛋白水平明显下调,且呈浓度和时间依赖性(P<0.05)。体外试验发现,Cr-1siRNA可有效抑制结直肠癌细胞集落生长和侵袭能力,且与浓度相关(P<0.05)。结论 Cr-1在结直肠癌细胞侵袭中起着重要的作用,采用Cr-1siRNA转染可抑制结直肠癌细胞侵袭。

Objective To study the effects of cripto-1（Cr-1）small interfering RNA（siRNA）on invasion of colon cancer cell. Methods After human colon cancer cell line SW480 were transfected by Cr-1 siRNA, the mRNA and protein of Cr-1 were determined by real time RT-PCR and Western Blot assay, respectively. The anchorage-independent growth was exmined by clon formation in soft agar,and invasion ability was evaluated by boyden chamber model.Results The siRNA could down-regulate the level of mRNA and protein of Cr-1 in a dose- and time- dependent manner. Suppression of of Cr-1 expression can inhibit anchorage-independent growth in a dose-dependent manner,and invasion ability of breast cancer cell.Conclusion Cr-1 gene might play an important role in development of human colon cancer,and down-regulation by Cr-1 siRNA could inhibit invasion of human colon cancer cell.