摘要
目的探讨吡柔比星对膀胱癌细胞增殖的影响及其作用机制。方法膀胱癌细胞T24和BIU-87分别用0.4、0.8、1.6、3.2mg/L的吡柔比星处理24、48、72h,用四甲基偶氮唑盐(MTT)检测细胞增殖,流式细胞术检测细胞凋亡率,qRT-PCR、RT-PCR检测磷脂酶Cε(PLCε)mRNA及凋亡调控基因Bcl-2mRNA的表达,蛋白质印迹法检测PLCε蛋白的表达。将膀胱癌细胞分为空白组、吡柔比星处理组、Ad-shPLCε处理组、吡柔比星联合PLCε干扰腺病毒载体(Ad-shPLCε)处理组,检测并比较各组细胞增殖和Bcl-2蛋白的表达量。结果在膀胱癌细胞T24、BIU-87中,吡柔比星对细胞的抑制率呈浓度、时间依赖性,即随着药物浓度增加、处理时间延长,细胞抑制率升高;吡柔比星促进T24和BIU-87细胞凋亡,并且抑制PLCε和Bcl-2表达。吡柔比星处理组、Ad-shPLCε处理组、Ad-shPLCε联合吡柔比星处理组均能抑制细胞增殖和Bcl-2表达,且Ad-shPLCε联合吡柔比星处理组的抑制作用高于吡柔比星处理组,差异有统计学意义(P<0.05)。结论吡柔比星能有效抑制膀胱癌细胞的增殖,可能与其抑制PLCε癌基因及下游Bcl-2基因的表达有关。
Objective To investigate the effects of pirarubicin on proliferation of bladder cancer cells and the related mechanism.Methods After bladder cancer cell lines T24 and BIU-87 were treated with 0.4,0.8,1.6,and 3.2mg/L pirarubicin for 24,48,and 72h,the cell proliferation was detected by MTT.Flow cytometry was used to examine the apoptosis of T24 and BIU-87 cells.qRT-PCR and RT-PCR were used to examine the mRNA expression of phospholipase Cε（PLCε）,Bcl-2in T24 and BIU-87 cell lines;the protein expression of PLCεin pirarubicin-treated cells was determined by Western blotting analysis.Bladder cancer cells were designed as blank group,pirarubicin treatment group,Ad-shPLCεtreatment group,and Ad-shPLCεplus pirarubicin treatment group;cell proliferation was observed and protein expression of Bcl-2 was examined and compared between different groups.Results Pirarubicin showed a dose- and time-dependent inhibitory effect against proliferation of T24 and BIU-87 cell lines.Moreover,pirarubicin promoted cell apoptosis in T24 and BIU-87 cells and suppressed the expression of PLCεand Bcl-2.Pirarubicin treatment group,Ad-shPLCεtreatment group,and Ad-shPLCεplus pirarubicin treatment group all had suppressed cell proliferation and Bcl-2expression,and pirarubicin plus Ad-shPLCεgroup exhibited significantly stronger inhibitory effects compared with pirarubicin treatment group（P0.05）.Conclusion Pirarubicin can effectively inhibit cell proliferation of bladder cancer cells,which may be through suppressing the expression of PLCεand Bcl-2.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2014年第7期708-713,共6页
Academic Journal of Second Military Medical University
基金
国家自然科学基金(81072086)
重庆市教委科学技术研究项目(KJ110305)~~
