CAJ | 학술논문

目的探讨3-甲基腺嘌呤(3-MA)自噬抑制剂联合吡柔比星(THP)对膀胱癌BIU-87细胞杀伤作用及机制.方法膀胱癌BIU-87细胞分为对照组、3-MA(10 mmol/L)组、THP(5 mg/L)组、3-MA(10 mmol/L)+ THP(5 mg/L)组.噻唑蓝(MTT)法检测细胞增殖;流式细胞术检测细胞凋亡;Western blot检测各组细胞中自噬蛋白微管相关蛋白1轻链3(LC3)-Ⅱ、Beclin-1以及凋亡蛋白半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3和Caspase-9的表达.结果药物作用24h后,各组细胞增殖率分别为(86.64±3.09)％、(63.70±5.62)％、(58.43±3.89)％、(37.55±4.17)％,其中3-MA+THP组细胞增殖下降最明显(P ＜0.05);3-MA与THP联合应用后,自噬蛋白LC3-Ⅱ、Beclin-1的表达明显减弱,而凋亡蛋白Caspase-3和Caspase-9的表达在各组中最强(P＜0.05).结论 3-MA抑制膀胱癌BIU-87细胞对化疗药物保护性自噬反应,增加膀胱癌细胞对THP细胞毒杀伤作用的敏感性.
목적 탐토3-갑기선표령(3-MA)자서억제제연합필유비성(THP)대방광암BIU-87세포살상작용급궤제.방법 방광암BIU-87세포분위대조조、3-MA(10 mmol/L)조、THP(5 mg/L)조、3-MA(10 mmol/L)+ THP(5 mg/L)조.새서람(MTT)법검측세포증식;류식세포술검측세포조망;Western blot검측각조세포중자서단백미관상관단백1경련3(LC3)-Ⅱ、Beclin-1이급조망단백반광안선천동안산특이성단백매(Caspase)-3화Caspase-9적표체.결과 약물작용24h후,각조세포증식솔분별위(86.64±3.09)％、(63.70±5.62)％、(58.43±3.89)％、(37.55±4.17)％,기중3-MA+THP조세포증식하강최명현(P ＜0.05);3-MA여THP연합응용후,자서단백LC3-Ⅱ、Beclin-1적표체명현감약,이조망단백Caspase-3화Caspase-9적표체재각조중최강(P＜0.05).결론 3-MA억제방광암BIU-87세포대화료약물보호성자서반응,증가방광암세포대THP세포독살상작용적민감성.
Objective To observe the cytotoxic effect of 3-methyladenine (3-MA) autophagy inhibitors combined with pirarubicin on bladder cancer cells (BIU-87),and its possible mechanism.Methods Four groups of bladder cancer cells at logarithmic phase were designed in this experiment:control,3-MA (10 mmol/L),pirarubicin (THP,5 mg/L) and 3-MA (10 mmol/L) +THP (5 mg/L).Optimal concentration and cell proliferation activity were determined by methyl thiazol tetrazolium (MTT)assay.The cell apoptosis rate was determined by annexin V-fluoresceine isothiocyanate (FITC)/propidium iodide (PI) flow cytometry.The expression of autophagy related proteins microtubule-associated pro-tein 1 light chain 3 (LC3)-Ⅱ and Beclin-1,and apoptins Caspase-3 and Caspase-9 was detected by Western blotting.Results The proliferation rate was (86.64 ± 3.09) ％,(63.70 ± 5.62) ％,(58.43 ± 3.89)％,and (37.55 ±4.17)％ in control,3-MA (10 mmol/L),THP (5 mg/L) and 3-MA (10 mmol/L) + THP (5 mg/L) groups respectively.The proliferation rate was decreased most significantly in 3-MA + THP group as compared with control group (P ＜ 0.05) at 24 h after treatment.The expression of LC3-Ⅱ and Beclin-1 was decreased in 3-MA + THP group,while that of Caspase-3 and Caspase-9 increased.The expression of LC3-Ⅱ,Beclin-1,Caspase-3 and Caspase-9 was increased in THP group.The expression of LC3-Ⅱ and Beclin-1 was decreased obviously in 3-MA + THP group,while that of Caspase-3 and Caspase-9 had the highest rate.Conclusion 3-MA inhibits protective autophagy response of bladder cancer cells to chemotherapy drugs,which can enhance the sensitivity of bladder cancer cells to pirarubicin.