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目的:研究Ras homology C(RhoC)在胃癌转移中的作用。方法:利用Vector NTI软件设计并构建RhoC 的小干扰RNA(siRNA)载体,利用脂质体介导法将一组RhoC siRNA分别转染胃癌SGC7901细胞。筛选出稳定抗性克隆;Western blot检测RhoC siRNA转染后的抑制效应,Cell Counting Kit-8检测RhoC siRNA转染细胞株的生长速度;MTT法检测转染细胞株对化疗药物的敏感性;损伤刮擦实验和TRANSWELL小室实验分别检测转染细胞株的迁移与侵袭能力。结果:在计算机辅助下成功地设计并构建了5个RhoC的小干扰RNA载体,Western blot显示其中一个RhoC的小干扰RNA RhoC-s362对 RhoC的表达有明显的抑制作用;尽管下调RhoC的表达对胃癌细胞的生长和药物敏感性无明显影响,但可显著抑制肿瘤细胞的迁移与侵袭。结论:RhoC siRNA RhoC-s362能够明显抑制胃癌SGC7901细胞中RhoC的表达,并进而抑制胃癌细胞的迁移与侵袭,提示RhoC可能在胃癌转移中发挥重要作用。
Objective: To investigate the role of Ras homology C (RhoC) in gastric cancer metastasis. Methods: Vector siRNA was designed and constructed to construct RhoC small interfering RNA (siRNA) vector. A group of RhoC siRNA was transfected into gastric cancer SGC7901 cells by liposome-mediated method. The stable resistant clones were screened out; the inhibitory effect of RhoC siRNA transfection was detected by Western blot, the growth rate of RhoC siRNA transfected cell lines was detected by Cell Counting Kit-8, the sensitivity of transfected cell lines to chemotherapeutic drugs was detected by MTT assay, Scratch experiments and TRANSWELL chamber experiments were performed to detect the migration and invasion ability of transfected cell lines. Results: Five siRNAs targeting RhoC were successfully designed and constructed by computer. Western blot showed that RhoC-s362, a small interfering RNA of RhoC, significantly inhibited the expression of RhoC. Although the expression of RhoC was down-regulated Gastric cancer cell growth and drug sensitivity had no significant effect, but significantly inhibited tumor cell migration and invasion. Conclusion: RhoC siRNA RhoC-s362 can significantly inhibit the expression of RhoC in gastric cancer SGC7901 cells, and then inhibit the migration and invasion of gastric cancer cells, suggesting that RhoC may play an important role in gastric cancer metastasis.