粉防己碱对膀胱癌BIU-87／ADM细胞株的耐药逆转作用及其机制  

Reversing mechanism of multidrug resistance of human bladder carcinoma drug-resistant cell line BIU-87/ADM by tetrandrine

作　　者：李永生[1] 刘锦裕[2] 黄玉清[2] 

机构地区：[1]福建医科大学附属协和医院泌尿外科,福州350001 [2]福建医科大学协和临床医学院

出　　处：《中华实验外科杂志》2011年第9期1526-1529,共4页Chinese Journal of Experimental Surgery

基　　金：福建省科技计划资助项目（2007F3036）

摘　　要：目的探讨粉防己碱（TET）对膀胱癌耐药株BIU-87／ADM多药耐药性（MDR）的逆转作用及其机制。方法应用CCK法观察不同浓度TET（1．0、1．5、2．0、2．5mg／L）对BIU-87和BIU-87／ADM细胞生长的抑制作用，并测定1mg／LTET逆转多药耐药的活性；Annexin—V与PI双染法流式细胞术分析1mg／LTET对细胞凋亡的影响；免疫荧光法和逆转录-聚合酶链反应（RT—PCR）分别检测1mg／LTET对P-gP蛋白和MDRl表达水平的影响；细胞免疫化学法和RT—PCR法检测1mg/LTET对BIU-87／ADM细胞Easpase-3和Survivin表达水平的影响。结果1．0mg／L无毒剂量下的TET显著增加阿霉素（ADM）对耐药株BIU-87／ADM的细胞毒性作用（P〈0．01），逆转指数（RV）为5．33，而对敏感株BIU-87无明显作用（P〉0．05），RF为1．33；1．0mg／LTET和1．5mg／LADM共同作用48h，能使BIU-87／ADM细胞凋亡率增加至（40．86±4．35）％，与单独应用ADM的（12．42±3．24）％比较差异有统计学意义（P〈0．01）。而BIU-87细胞凋亡率为（61．23±5．46）％，与单独应用ADM的（57．79±4．73）％比较差异无统计学意义（P〉0．05）。TET能明显抑制mdrlmRNA和P-gP蛋白在BIU-87／ADM的表达；与单用ADM比较，TET和ADM联用能使BIU-87／ADM细胞株中Caspase．3的表达水平明显升高（P〈0．01），Survivin的表达水平显著降低（P〈0．01）。结论粉防己碱能逆转BIU-87／ADM细胞的多药耐药性，这一作用可能与粉防己碱抑制P-gP的表达和增强化疗药诱导细胞凋亡有关。Objective To Investigate the reversal effect and the mechanism of tetrandrine （TET） on human bladder carcinoma cell line BIU-87/ADM. Methods The effects of TET （ 1.0, 1.5, 2.0, 2. 5 mg/L） on the growth of BIU-87 and BIU-87/ADM cells were measured by CCK assay. The potential of 1 mg/L TET to reverse multidrug resisitance （MDR） was also determined by CCK assay. The apoptosis of cell lines induced by 1 mg/L TET was evaluated by Annexin-V and PI followed by flow cytometry （FCM）. The effects of 1 mg/L TET on the expression of P-gp and MDR1 were separately measured by immunofluo- rescence and reverse transcription-polymerase chain reaction （RT-PCR） , and the expression of Caspase-3 and Survivin was analyzed by immunocytochemical technique and RT-PCR. Results TET at the concentra- tion of 1. O mg/L which had no obvious cytotoxicity to the BIU-g7 and BIU-gT/ADM cells increased the cy- totoxicity of adriamycin （ADM） in BIU-87/ADM ceils greatly, with a reversal fold of 5.33, but had no significant effect on BIU-87 cells, with a reversal fold of 1.33. The ADM-induced apoptosis rate in BIU-87 and BIU-87/ADM cells was （57.79 ± 4.73 ） % and （ 12.42 ± 3.24） % respectively. After treatment of BIU-87/ADM cells with 1.0 mg/L TET, the apoptosis rate was increased to （40. 86 ±4. 35）%, but it made no difference in BIU-87 ceils. The expression of mdrl and P-gp was suppressed by TET in BIU-87/ADM cells. Compared to the ADM group, Caspase-3 was more highly expressed in BIU-87/ADM cells of ADM ＋TET group. However, Survivin had the opposite result. Conclusion TET can reverse the MDR of the human bladder carcinoma BIU-87/ADM cell line, which may be associated with the decreased expression of P-gp and increased apoptosis.

关 键 词：粉防己碱 阿霉素 膀胱癌 抗药性 

分 类 号：R285.5[医药卫生—中药学]