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作 者:商延芳[1] 金月玲[1] 徐佳佳[1] 李楠[1] 黄培林[1]
出 处:《现代生物医学进展》2008年第1期19-21,26,共4页Progress in Modern Biomedicine
基 金:国家科学自然基金(NO.30471937)
摘 要: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.
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