中文摘要：

背景与目的：蛋白酶体抑制剂硼替佐米是新型抗癌药物，体外对多种恶性肿瘤细胞均具有良好的疗效。本实验研究硼替佐米与紫杉醇单独及联合应用对人卵巢癌细胞株SKOV3的存活率和凋亡率的影响，并对糖原合成酶激酶-3β（GSK-3β）和髓样白血病-1基因（Mcl-1）在其诱导细胞凋亡中的作用机制进行探讨，为临床治疗卵巢癌提供理论依据。方法：用50nmol/L硼替佐米、90nmol／L紫衫醇，或50nmol／L硼替佐米联合90nmol/L紫衫醇分别作用于SKOV3细胞12、24、36、48及72h后，四甲基偶氮唑蓝（MTT）比色法测定细胞增殖活性，并计算细胞存活率，Annexin-V／PⅠ法流式细胞仪检测细胞凋亡率，蛋白质免疫印迹法（Western blot）检测相关蛋白表达水平。结果：50nmol／L硼替佐米和90nmol／L紫衫醇联合作用于SKOV3细胞，12、24、36、48及72h时间点细胞存活率分别为（65．2±5．8）％，（58．3±14．4）％，（35．3±5．0）％．（19．2±1．5）％和（11．4±2．5）％，与紫杉醇单用组比较，细胞生长抑制增强，差异有显著性（P〈0．05）。药物单独或联合作用细胞24h后，紫杉醇单用组、硼替佐米单用组和两者联用组细胞凋亡率分别为（14．7±0．5）％、（15．1±0．8）％和（20．5±0．7）％，硼替佐米与紫杉醇联用组凋亡率显著增高（P〈0．05）。药物处理细胞后，经免疫印迹法检测p-GSK-3β和Mcl-1蛋白表达水平，硼替佐米与紫杉醇联用组P—GSK-3β和Mcl-1表达水平降低最为明显，分别为正常细胞组表达水平的（19．3±0．4）％和（31．6±3．1）％，差异均宵显著性（P〈0．05）。硼替佐米和紫杉醇单用组中p-GSK-3β表达水平有所降低，分别为正常细胞组的（78．7±1．2）％和（85．1±1．6）％，Mcl-1表达水平分别为对照组的（68．2±4．5）％和（57．0±4．1）％，差异均具显著性（P〈0．05）。结论：?

英文摘要：

Background and purpose： Proteasome inhibitors such as bortezomib, represent an interesting new class of potential anticancer drugs. In the present study,we explored the sensitivity of ovarian cancer cell line SKOV3 to paclitaxel, proteasome inhibitors and their combination, and also studied the involvement of GSK-3β/Mcl-1 signaling pathway in the regulation of apoptosis induced by those agent. Methods： Methyl thiazolyl tetrazolium （MTT） assay was applied to examine the cell viability, Annexin-V/PI apoptosis detection kit was used to determine the apoptosis rate of different groups, and western blot assay was introduced to evaluate the expression levels of phosphorylated GSK-3β and Mcl-1. Results： In the MTT assay, the cell viability ratios of combination group at serial time points from 12 to 72 hr were （65.2±5.8）%, （58.3±14.4）%, （35.3±5.0）%, （19.2±1.5）% and （11.4±2.5）%, and there were significant differences as compared to the treatment of paclitaxel alone （P〈0.05）. After drug treatments, apoptosis rates of paclitaxel group, bortezomib group and combination group were （14.7±0.5）%, （15.1±0.8）% and （20.5±0.7）%,respectively. The rate of combination group was much higher than that in the other two groups （P〈0.05）. Western blot assay showed that after the treatment of paclitaxel and bortezomib combination, phosphorylated GSK- 3β and Mcl-1 were significantly down-regulated （P〈0.05）. Conclusions： The cytostatic effect of paclitaxel could be increased significantly by the combination of proteasome inhibitors like bortezomib and paclitaxel. And the GSK- 3β/Mcl-1 signaling pathway may play an important role in the molecular mechanism of apoptosis induced by the combination treatment.