摘要
目的 探讨ABT-737联合多西他赛对人前列腺癌PC-3细胞的协同杀伤效应及相关机制.方法 将培养的PC-3细胞接种在96孔板,分别给予100、200、400、800 nmol/L ABT-737和5、10、20、30 nmol/L多西他赛单药处理24、48 h；联合用药组使用400 nmol/L ABT-737联合5、10、20、30 nmol/L多西他赛处理48 h.MTT法检测ABT-737和（或）多西他赛对PC-3细胞生长的影响；细胞形态学、流式细胞仪等检测细胞凋亡及细胞周期变化；Western blot、比色法分别检测凋亡相关蛋白Bcl-2、Bax、Bcl-xL和Mcl-1的表达以及caspase-3活性变化.结果 药物作用48 h后,ABT-737（400 nmol/L）＋多西他赛（20 nmol/L）联合作用后的细胞生存率为19.7％±3.2％,低于多西他赛（20 nmol/L）组（44.2％±4.4％）（t=4.45,P＜0.05）和ABT-737（400 nmol/L）组（93.2％±1.8％）,二者有协同抑瘤作用（CI=0.8）.联合药物组的sub-G1期细胞（38.5％±3.7％）明显高于多西他赛组（11.2％±1.3％）和ABT-737组（2.6％±0.4％）（F=50.88,P＜0.05）,多西他赛组导致G2/M期细胞周期阻滞.联合药物组明显抑制Bcl-2、Bcl-xL和Mcl-1蛋白表达（F=369.53、57.89、32.77；均P＜0.05）；caspase-3活性（419.7％±15.6％）增加（F=207.33,P＜0.05）.结论 ABT-737联合多西他赛可以通过诱导细胞凋亡发挥协同作用、抑制前列腺癌细胞生长,该机制可能与细胞周期阻滞,Bcl-2、Bcl-xL、Mcl-1和caspase-3表达及活性变化有关.
Objective To investigate the synergistical killing effect of docetaxel combined with ABT-737 on human prostate cancer cell line PC-3 by inducing apoptosis and further to determine the mechanism underlying such effect.Methods PC-3 cells were treated with various concentrations of docetaxel or（and）ABT-737.Cell viability was determined using MTT assay.Apoptosis was assessed by fluorescence microscopy analysis of cells with condensed and segmented nuclei following staining with 4＇,6-diamidino-2-phenylindole（DAPI）.Cellular DNA was stained with propidium iodide and flow cytometric analysis was performed to analyze the cell cycle distribution.Bcl-2,Bax,Bcl-xL and Mcl-1 protein changes were detected by Western blot.The activity of caspase-3 was measured using a colorimetric assay.Results Docetaxel（20 nmol/L）combination with ABT-737（400 nmol/L）for 48 hours,the cell viability was decreased to 19.7% ±3.2% to compare with 44.2% ±4.4%（t =4.45）of docetaxel and 93.2% ± 1.8% of ABT-737 separately and there was a synergistic effect between the two drugs（CI =0.8）.Apoptosis rate of the combination group was higher than other two drugs.Docetaxel increased the cell number arrested in G2/M phase compared with control group（P 〈 0.05）,but the combination treatment resulted in a significant arrest in the G0/G1 phase.The combination treatment could significantly reduced the Bcl-2,Bcl-xL and Mcl-1 expression（F =369.53,57.89 and 32.77,all P 〈 0.05）and enhanced the activity of caspase-3（419.7 % ± 15.6%）（F =207.33,P 〈 0.05）.Conclusions The combination of ABT-737 with docetaxel can synergistically inhibit the proliferation of PC-3 cells through inducing apoptosis,which may be associated with cell cycle arrest,down-regulation of Bcl-2,Bcl-xL and Mcl-1 expression and activation of caspase-3.
出处
《中华外科杂志》
CAS
CSCD
北大核心
2012年第2期161-165,共5页
Chinese Journal of Surgery
基金
国家自然科学基金资助项目（81001146）
