DLC-1基因表达对结肠癌细胞侵袭转移能力的影响

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

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。

Objective： To investigate the effection of DLC-1 gene on migration of human colorectal cancer LoVo cell line. Methods： Short hairpin RNA（ ShRNA ） was desighed to inhibit DLC-1 gene expression and DLC-1 shRNA recombinant was cloned into pGCsi-U6/Neo plasmid, and the product was transfected into DLC-1 positive LoVo cells with Lipofectamine 2000. The expression level of DLC-1mRNA 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, After 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 technique 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.