白藜芦醇联用紫杉醇对食管癌EC109细胞凋亡的影响

Influence of Resveratrol Combined with Paclitaxel on Apoptosis of Esophageal Cancer Cells EC109

目的探讨白藜芦醇联合紫杉醇对食管癌EC109细胞凋亡的影响及其机制。方法实验分为白藜芦醇组、紫杉醇组、白藜芦醇联用紫杉醇组以及空白对照组。采用MTT法检测各个用药组对EC109细胞活力的影响;采用流式细胞仪检测各个用药组对细胞凋亡的影响;采用罗丹明123外排实验检测各个用药组细胞的外排功能;采用HPLC法检测各个用药组细胞内紫杉醇的含量。结果MTT实验结果显示,与单用紫杉醇相比,白藜芦醇减弱了紫杉醇对细胞增殖的抑制作用;克隆形成实验结果显示,与单用紫杉醇组相比,白藜芦醇和紫杉醇联用后,EC109细胞的克隆形成能力明显增强;Hochest 33258实验结果显示,白藜芦醇和紫杉醇联用后,相对于单用紫杉醇组,EC109细胞的凋亡小体以及核裂解数减少;流式细胞术分析细胞凋亡的结果显示,白藜芦醇和紫杉醇联用后,与单用紫杉醇组相比,凋亡率明显下降。HPLC和罗丹明123外排实验结果证明,白藜芦醇和紫杉醇联用后,与单用紫杉醇组相比,紫杉醇在细胞内的含量明显降低。结论白藜芦醇具有拮抗紫杉醇诱导的细胞凋亡作用,其机制可能与白藜芦醇增加了细胞的P-gp蛋白的表达、减少了紫杉醇在细胞内的滞留从而导致紫杉醇的药效减弱有关。

Objective To investigate the effect of resveratrol combined with paclitaxel on the apoptosis of esophageal cancer cells EC109.Methods EC109 cells were divided into resveratrol group,paclitaxel group,resveratrol+paclitaxel group and control group.MTT assay was used to detect the effect on the viability of EC109 cells,flow cytometry was used to detect the effect on the apoptosis of the cells,Rhodamine 123 efflux assay was used to detect the efflux function of the cells,and HPLC method was used to detect the accumulation of intracellular paclitaxel in each group.Results The results of the MTT assay showed that the combined use of resveratrol and paclitaxel weakened the inhibitory effect of paclitaxel on cell proliferation compared with using paclitaxel alone.The results of colony formation assay showed that the colony formation ability of EC109 cells in the combined group of resveratrol and paclitaxel was enhanced significantly in comparison with paclitaxel group.Hochest 33258 assay showed that the number of apoptotic bodies and nuclear lysis in EC109 cells of the combined group was decreased as compared with that in the paclitaxel group.Flow cytometry showed that the apoptotic rate of the combined group was significantly decreased in comparison with the paclitaxel group.The results of HPLC and Rhodamine 123 efflux assays proved that the accumulation of intracellular paclitaxel in the combined group was significantly reduced in comparison with the paclitaxel group.Conclusion Resveratrol can antagonize the effect of paclitaxel on inducing apoptosis,and the mechanism is probably related with the increase of the expression of Pgp and with the decrease of the accumulation of intracellular paclitaxel in the EC109 cells,which results into the decrease of the pharmacological action of paclitaxel.