DLC-1基因表达对结肠癌细胞侵袭转移能力的影响

目的:研究DLC-1基因对结肠癌细胞侵袭迁移能力的影响.方法:将DLC-1 shRNA(短发夹状RNA,short hairpin RNA)序列克隆到质粒pGCsi-U6/Neo载体,采用脂质体介导的转染方法将构建的DLC-1 shRNA表达质粒转入结肠癌细胞系LoVo细胞.采用RT-PCR技术和Western Blot技术分别检测LoVo细胞中DLC-1mRNA和蛋白表达水平的变化.Transwell小室人工重组基底膜侵袭转移实验观察LoVo细胞侵袭迁移能力的改变.结果:结肠癌细胞系LoVo细胞表达DLC-1分子.所构建质粒表达载体能有效地干扰LoVo细胞DLC-1 mRNA和蛋白质表达水平;Transwell小室人工重组基底膜侵袭转移实验结果显示,转染后LoVo细胞侵袭转移能力明显增强(p＜0.05).结论:结肠癌细胞系LoVo细胞表达DLC-1基因,应用RNAi技术可特异性降低其表达.DLC-1的表达水平与结肠癌细胞侵袭转移相关.

Effect of DLC-1 Gene on Invasion and Migration of Human Colorectal Cancer Cell Line

Objective:To investigate the effection of DLC-1 gene on migration of human colorectal cancer LoVo cell line.Meth- ods:Short hairpin RNA(ShRNA)was desighed to inhibit DLC-1 gene expression and DLC-1 shRNA recombinant was cloned into pGC- si-U6/Neo plasmid,and the product was transfected into DLC-1 positive LoVo cells with Lipofectamine 2000.The expression level of DLC-I mRNA was detected by RT-PCR and the expression level of DLC-1 protein was measured by Western Blot.The effects of DLC-1 on invasion and mobility abilities of LoVo cells in vitro were explored by transwell chamber.Results:DLC-1 gene could be detected in LoVo cells.The DLC-1 shRNA plasmid expression vector we made could knock down DLC-1 expression on LoVo cells effectively.Af- ter transfected into the DLC-1 shRNA plasmid expression vector,the invasive and metastatic potention of human colorectal cancer LoVo cell line was increased significantly(p<0.05).Conclusion:The human colorectal cancer cell line LoVo can express DLC-1.The tech- nique of RNAi can knock down the DLC-1 expression.The expression level of DLC-1 was associated with the ability of invasive and metastasis of human colorectal cancer LoVo cell line.