TRPM7在膀胱癌细胞增殖与凋亡中的调控作用

目的 探讨薄荷醇受体7（transient receptorpotential melastatin 7,TRPM7）在膀胱癌细胞株T24细胞增殖与凋亡中的调控作用及分子机制.方法 采用RT-PCR及Western blot检测T24细胞株中TRPM7 mRNA及蛋白的表达;分别采用通道阻滞剂及基因沉默的方法阻断TRPM7离子通道的功能,MTT法检测细胞存活率,流式细胞术检测细胞周期分布及细胞凋亡率,Western blot检测Cdk4、Cdk6及Cyto C的表达.结果 RT-PCR及Western blot证实T24细胞株中存在TRPM7 mRNA及蛋白的表达;采用基因沉默及通道阻滞剂阻断TRPM7的功能后,T24细胞存活率分别下降了56.48％和54.87％,处于G0/G1期的细胞随阻滞剂浓度增加而显著增加,细胞凋亡率亦随之增加并呈浓度依赖性,与对照组比较差异均有统计学意义（P＜0.05）.Western blot显示阻断TRPM7后,T24细胞Cdk4、Cdk6的表达减少,而Cyto C的表达增加.结论 TRPM7可促进T24细胞增殖,抑制细胞凋亡.这一过程可能通过调节Cdk4、Cdk6及Cyto C的表达来实现.阻断TRPM7的功能,能够抑制T24细胞增殖,促进细胞凋亡,可能为临床治疗膀胱癌提供新的靶点.

Effect of TRPM7 on the proliferation and apoptosis in bladder cancer T24 cells

Objectives To investigate the effects of transient receptor potential melastatin 7 （TRPM7） on the proliferation and apoptosis in T24 bladder cancer cell lines and the underlying molecular mechanisms.Methods Expression of TRPM7 mRNA and protein in T24 cell line were detected with RT-PCR and Western blot; channel blockers and gene silencing were used to block the function of TRPM7,MTT assay was used to detect cell viability,flow cytometry was used to analyze cell cycle distribution and apoptosis rate,Western blot was used to detect the expression of Cdk4,Cdk6 and Cyto C.Results RT-PCR and Western blot analysis confirmed over-expression of TRPM7 mRNA and protein in T24 cell lines ; after using gene silencing and channel blockers to block the function of TRPM7,the viability of T24 cell decreased by 56.48％ and 54.87％ respectively,T24 cells at G0 / G1 stage and the apoptosis rate increased significantly in a dose-dependent manner.Compared with the control group,the differences were statistically significant （p ＜ 0.05）.Western blot showed that after blocking TRPM7 expression,Cdk4,Cdk6 in T24 cells decreased,while the expression of Cyto C increased.Conclusions TRPM7 ion channel can promote cell proliferation and inhibit cell apoptosis in T24 cells.This process may achieve by regulating the expression of Cdk4,Cdk6 and Cyto C.Blocking TRPM7 function,T24 can inhibit cell proliferation and induce apoptosis,TRPM7 may provide a new target for the clinical treatment of bladder cancer.