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目的探讨m TOR抑制剂依维莫司与紫杉醇共同抑制耐阿霉素乳腺癌(MCF-7/ADR)细胞株生长的机制。方法在培养耐阿霉素乳腺癌细胞株(MCF-7/ADR)中加入一定量的依维莫司和紫杉醇,采用MTT法对加入依维莫司和紫杉醇的细胞IC50含量进行检测,然后根据检测结果来进行依维莫司与紫杉醇浓度的设置,并计算联合用药的效果。采用流式细胞术对MCF-7/ADR细胞在不同药组作用下48h后的凋亡率进行检测。采用Western blot方法对各组细胞内的m TOR、p-m TOR、PI3K、p-PI3K、AKT、p-AKT、4EBP1和p-4EBP1蛋白表达情况进行检测。结果依维莫司和紫杉醇抑制MCF-7/ADR细胞增殖的半数抑制浓度(IC50)分别为32.58μg/ml和9.59μg/ml,两者合用抗增殖作用大于紫杉醇或依维莫司单用,组合指数均小于0.9。与只用紫杉醇对MCF-7/ADR细胞进行处理相比,联合使用依维莫司和紫杉醇能更好地诱导MCF-7/ADR细胞凋亡。Western blot结果显示,紫杉醇和依维莫司合用可明显加强对m TOR、p-m TOR、PI3K、p-PI3K、p-AKT、4EBP1和p-4EBP1表达的抑制作用。结论依维莫司合用紫杉醇,可增强对MCF-7/ADR细胞增殖抑制和凋亡的诱导作用,其机制可能是下调m TOR信号通路及上、下游信号蛋白表达和磷酸化。
Objective To investigate the mechanism of mTOR inhibitor everolimus co-inhibiting the growth of adriamycin-resistant breast cancer (MCF-7 / ADR) cell line. Methods A certain amount of everolimus and paclitaxel were added into adriamycin resistant breast cancer cell line (MCF-7 / ADR). The IC50 of cells treated with everolimus and paclitaxel was determined by MTT method. The test results were used to determine the concentration of everolimus and paclitaxel, and the effect of combination therapy was calculated. Flow cytometry was used to detect the apoptosis rate of MCF-7 / ADR cells after 48h under different drug groups. The expression of mTOR, p-m TOR, PI3K, p-PI3K, AKT, p-AKT, 4EBP1 and p-4EBP1 in each group were detected by Western blot. Results The median inhibitory concentration (IC50) of everolimus and paclitaxel on MCF-7 / ADR cell proliferation was 32.58μg / ml and 9.59μg / ml, respectively. The combined effect of both of them was more effective than paclitaxel or everolimus, The composite index is less than 0.9. The combination of everolimus and paclitaxel induced better apoptosis in MCF-7 / ADR cells compared to paclitaxel-only treatment of MCF-7 / ADR cells. Western blot results showed that combination of paclitaxel and everolimus could obviously inhibit the expression of mTOR, p-mTOR, PI3K, p-PI3K, p-AKT, 4EBP1 and p-4EBP1. Conclusion Combination of everolimus and paclitaxel can enhance the induction of apoptosis and proliferation of MCF-7 / ADR cells. Its mechanism may be down-regulation of mTOR signaling pathway and up-and down-stream signal protein expression and phosphorylation.