1504-siRNA对表达EphA3的肿瘤细胞HGC27的抑制作用

Inhibition Effect of 1504-siRNA on EphA3 Expressed HGC27 Cells

目的:研究原癌基因EphA3的小干扰RNA(siRNA)1504-siRNA对表达EphA3的胃癌细胞HGC27的增殖抑制作用。方法:将携带1504-siRNA的质粒用Vigorous转染试剂瞬时转染HGC27细胞,48 h后收集蛋白,用Western印迹检测EphA3及AKT信号通路中的蛋白分子表达;36 h后收集转染细胞,进行MTT、平板克隆形成和软琼脂克隆形成实验。结果:1504-siRNA能抑制HGC27细胞的EphA3蛋白水平,抑制其增殖、平板克隆形成和软琼脂克隆的形成,抑制pAKT、pmTOR、p-c-Raf、p-4ebp1等信号分子的表达。结论:1504-siRNA可能通过抑制AKT信号通路,而抑制HGC27细胞的增殖、存活能力和恶性程度,它可以作为肿瘤治疗的候选药物。

Objective： To investigate the inhibition effects of the small interfering RNA（siRNA）, 1504-siRNA, targeting EphA3, on proliferation of gastric cancinoma cells HGC27. Methods： After plasmid carrying 1504-siRNA was transfected into HGC27 cells, protein were collected and then the protein levels of EphA3, AKT signal molecules were tested by Western blot, furthermore, cells were collected and MTT, plate clonogenicity and soft agar clonogenicity experiment were accomplished. Results： 1504-siRNA reduced the level of EphA3, pAKT, pmTOR, p-c-Raf, and p-4ebpl, et al. Furthermore, it surpressed the abilities of cell proliferation, plate clonogenicity and soft agar clonogenicity of EphA3 highly expressed tumor cells. Conclusion： 1504-siRNA should be a promising drug for cancer therapy for it can inhibit the proliferation, survival, and malignancy of EphA3 highly expressed tumor cells, which may be the consequences of suppression of AKT signal pathway.