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目的 RNA干扰使RhoA基因沉默后对结肠癌LoVo细胞在裸鼠体内生长的影响。方法构建靶向RhoA基因的shRNA真核表达载体,转染LoVo细胞,筛选RhoA基因沉默的稳定克隆。采用Western blot法检测RhoA蛋白的表达水平。将稳定转染RhoA shRNA表达载体、转染空载体和未转染的LoVo细胞分别接种于裸鼠皮下,4周后测量瘤体的体积和重量;免疫组化检测肿瘤组织RhoA,VEGF和MMP-2蛋白的表达。结果 RhoA shRNA转染组瘤体的体积与瘤重明显低于其他两对照组(P<0.01)。免疫组化结果显示,RhoA shRNA染组瘤体组织RhoA,VEGF和MMP-2蛋白表达均较两对照组显著降低(P<0.01)。结论沉默RhoA可以抑制结肠癌LoVo细胞移植瘤的生长。
Objective To investigate the effect of RhoA gene silencing RNAi on the growth of colon cancer LoVo cells in nude mice. Methods shRNA eukaryotic expression vector targeting RhoA gene was constructed and transfected into LoVo cells to screen for stable clones with silencing of RhoA gene. Western blot was used to detect the expression of RhoA protein. The RhoA shRNA expression vector was stably transfected into the nude mice. The empty vector and the untransfected LoVo cells were inoculated subcutaneously in nude mice respectively. After 4 weeks, the volume and weight of the tumor were measured. The expressions of RhoA, VEGF and MMP- 2 protein expression. Results The volume and tumor weight of RhoA shRNA transfected group were significantly lower than those of the other two control groups (P <0.01). The results of immunohistochemistry showed that the expression of RhoA, VEGF and MMP-2 in RhoA shRNA group was significantly lower than that in the two control groups (P <0.01). Conclusion Silencing RhoA can inhibit the growth of colon cancer LoVo cell xenografts.