冬凌草甲素对膀胱癌细胞凋亡作用及其分子机制研究

Study on molecular mechanism of oridonin promoting apoptosis of bladder cancer

目的:研究冬凌草甲素对膀胱癌的抗肿瘤作用及其分子机制。方法:购买人膀胱癌细胞T24,采用CCK8实验、集落形成、Transwell迁移实验和Annexin V-FITC凋亡分析检测冬凌草甲素对T24细胞的增殖、迁移、集落形成和凋亡的影响。采用Western blotting或RT-PCR检测冬凌草甲素对T24细胞凋亡相关蛋白、瞬时受体电位M7通道(TRPM7)和信号分子表达的影响。结果:冬凌草甲素处理膀胱肿瘤T24细胞后,T24细胞的增殖率、迁移的细胞数量、菌落形成率显著下降。实验组的p53、裂解的caspase-3蛋白水平均高于对照组,实验组TRPM7、p-AKT的表达水平均低于对照组,且其表达水平变化均呈浓度依赖性。结论:冬凌草甲素对T24细胞有明显的抗增殖和抗迁移作用,机制可能是通过靶向失活TRPM7、下调AKT、抑制AKT信号通路实现的。

Objective To investigate the antitumor effect of oridonin on bladder cancer(BC),and its molecular mechanism.Methods CCK 8 assay,Annexin V-FITC apoptosis assay,colony formation and Transwell migration assays were used to detect the effects of Oridonin on the proliferation,migration,apoptosis and colony formation of T24,respectively.Western blotting or RT-PCR was used to detect the effect of oridonin on the expression of apoptosis-related proteins,transaminase transient receptor potential channel 7(TRPM7)and signaling molecules in T24 cells.Results After treatment of bladder tumor T24 cells with oridonin,the proliferation rate,the number of migrated cells and the colony formation rate of T24 cells were significantly decreased.The protein levels of p53 and cleaved caspase-3 in the experimental group were higher than those in the control group,and the expression levels of TRPM7 and p-AKT in the experimental group were lower than those in the control group,and their expression levels changed in a concentration-dependent manner.Conclusion Oridonin has obvious anti-proliferation and anti-migration effects on T 24 cells,and the mechanism may be achieved by inhibiting of TRPM7,and inactivating AKT signaling pathway.