国际泌尿系统杂志
國際泌尿繫統雜誌
국제비뇨계통잡지
INTERNATIONAL JOURNAL OF UROLOGY AND NEPHROLOGY
2014年
5期
637-641
,共5页
膀胱肿瘤%细胞增殖%细胞凋亡
膀胱腫瘤%細胞增殖%細胞凋亡
방광종류%세포증식%세포조망
Urinary Bladder Neoplasms%Cell Proliferation%Apoptosis
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.
目的 探討薄荷醇受體7(transient receptorpotential melastatin 7,TRPM7)在膀胱癌細胞株T24細胞增殖與凋亡中的調控作用及分子機製.方法 採用RT-PCR及Western blot檢測T24細胞株中TRPM7 mRNA及蛋白的錶達;分彆採用通道阻滯劑及基因沉默的方法阻斷TRPM7離子通道的功能,MTT法檢測細胞存活率,流式細胞術檢測細胞週期分佈及細胞凋亡率,Western blot檢測Cdk4、Cdk6及Cyto C的錶達.結果 RT-PCR及Western blot證實T24細胞株中存在TRPM7 mRNA及蛋白的錶達;採用基因沉默及通道阻滯劑阻斷TRPM7的功能後,T24細胞存活率分彆下降瞭56.48％和54.87％,處于G0/G1期的細胞隨阻滯劑濃度增加而顯著增加,細胞凋亡率亦隨之增加併呈濃度依賴性,與對照組比較差異均有統計學意義(P＜0.05).Western blot顯示阻斷TRPM7後,T24細胞Cdk4、Cdk6的錶達減少,而Cyto C的錶達增加.結論 TRPM7可促進T24細胞增殖,抑製細胞凋亡.這一過程可能通過調節Cdk4、Cdk6及Cyto C的錶達來實現.阻斷TRPM7的功能,能夠抑製T24細胞增殖,促進細胞凋亡,可能為臨床治療膀胱癌提供新的靶點.
목적 탐토박하순수체7(transient receptorpotential melastatin 7,TRPM7)재방광암세포주T24세포증식여조망중적조공작용급분자궤제.방법 채용RT-PCR급Western blot검측T24세포주중TRPM7 mRNA급단백적표체;분별채용통도조체제급기인침묵적방법조단TRPM7리자통도적공능,MTT법검측세포존활솔,류식세포술검측세포주기분포급세포조망솔,Western blot검측Cdk4、Cdk6급Cyto C적표체.결과 RT-PCR급Western blot증실T24세포주중존재TRPM7 mRNA급단백적표체;채용기인침묵급통도조체제조단TRPM7적공능후,T24세포존활솔분별하강료56.48％화54.87％,처우G0/G1기적세포수조체제농도증가이현저증가,세포조망솔역수지증가병정농도의뢰성,여대조조비교차이균유통계학의의(P＜0.05).Western blot현시조단TRPM7후,T24세포Cdk4、Cdk6적표체감소,이Cyto C적표체증가.결론 TRPM7가촉진T24세포증식,억제세포조망.저일과정가능통과조절Cdk4、Cdk6급Cyto C적표체래실현.조단TRPM7적공능,능구억제T24세포증식,촉진세포조망,가능위림상치료방광암제공신적파점.
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.
