Sorafenib联合紫杉醇不同给药顺序作用于人肝癌细胞BEL-7402的效应

Schedule-dependent effects of sorafenib in combination with paclitaxel on human hepatocellular carcinoma cell line BEL-7402

背景与目的:Sorafenib是一种多靶点的抗肿瘤药物,其治疗原发性肝癌的疗效已经初步得到证实。本研究应用Sorafenib与紫杉醇(paclitaxel,TAX)联合,观察两药不同的给药顺序作用于人肝癌细胞BEL-7402的不同效应,初步探讨产生这种差异的机制。方法:采用MTT法检测Sorafenib与紫杉醇单药作用于BEL-7402细胞的IC50。流式细胞术检测Sorafenib与紫杉醇不同给药顺序作用后,BEL-7402细胞周期和凋亡率的变化。Western blot检测不同给药顺序作用后的BEL-7402细胞Bcl-2的表达情况。结果:Sorafenib和紫杉醇单药作用于BEL-7402细胞48h的IC50分别为(2.43±0.32)μg/mL、(1.89±0.72)μg/mL。分析Sorafenib、紫杉醇单药,先用紫杉醇再加入Sorafenib,先用Sorafenib再加入紫杉醇及紫杉醇与Sorafenib同时作用后BEL-7402细胞周期及凋亡率的变化,结果显示:①Sorafenib主要将细胞阻滞在S期,而紫杉醇主要将细胞阻滞在G2-M期。②先用紫杉醇再给予Sorafenib组细胞S期及G2-M期均有延长,且较其他组获得较高的凋亡率(36.43±2.29)%(P<0.01)。Western blot显示先用紫杉醇再给予Sorafenib组BEL-7402细胞的Bcl-2表达的水平最低。结论:Sorafenib联合紫杉醇不同给药顺序作用于BEL-7402细胞,先应用紫杉醇诱导再序贯给予Sorafenib时细胞凋亡率更高;产生这种不同效应的可能机制为两种药物作用于不同的细胞周期,以及不同给药方法后的Bcl-2的表达水平不同。

Background and Objective： Sorafenib is a multi-targeted antitumor drug. The monotherapy efficacy of sorafenib is relatively low. This study was to evaluate the schedule-dependent effect of sorafenib in combination with paclitaxel （TAX） on human hepatocellular carcinoma cell BEL-7402, and explore the underlying mechanism. Methods. BEL-7402 cells were treated with sorafenib or paclitaxel alone or in three different schedules： sorafenib was give prior to, after, or simultaneously with paclitaxel. The haft maximal inhibitory concentration （IC50） of sorafenib and paclitaxel was estimated by MTT. Alteration of cell cycle and apoptosis were analyzed by flow cytometry. The protein level of Bcl-2 in BEL-7402 cells was measured by western blot. Results： At 48 h, the IC50 of sorafenib and paclitaxel for BEL-7402 cells was （2.43±0.32） μg/mL and （1.89±0.72） μg/mL, respectively. Sorafenib caused cell cycle arrest at S phase, while paclitaxel blocked cells at GJM phase. S and GJM phases were extended and a higher apoptotic rate （36.43±2.29）% was induced when sorafenib was given after paclitaxel in Comparision with other groups （P〈0.01）. The protein level of Bcl-2 was the lowest in BEL-7402 cells treated with sorafenib after paclitaxel. Conclusions：Administration of sorafenib after paclitaxel induces a higher apoptotic rate in BEL-7402 cells than administration before or simutanously with paclitaxel. This is probably due to that the two drugs act on different cell cycle phases and the expression of Bcl-2 might be involved.