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目的:探讨在不同浓度阿霉素作用下,肺癌细胞株A549存活率发生反向变化的可能机制。方法:在阿霉素作用于A549细胞24小时后,采用MTT法检测细胞存活情况的变化;用实时定量RT-PCR和蛋白印迹法测定survivin的mRNA和蛋白水平表达;免疫印迹检测p21蛋白表达水平,以及流式细胞术检测药物作用后细胞周期变化。进一步用羟基脲阻滞细胞分化于G1期,观察阿霉素对细胞的作用情况。结果:在不同浓度梯度的阿霉素作用下(由低至高),细胞存活率出现先下降后上升的反弹趋势(P<0.01),并伴随survivin分子的高表达,同时,药物作用下细胞分化更倾向阻滞于G1期。结论:高浓度阿霉素作用下细胞分化多阻滞于G1期,并诱导出现survivin分子的高表达,从而增强了对细胞的保护作用是细胞存活率反向升高的可能原因。
Objective: To explore the possible mechanism of the reverse change of the survival rate of lung cancer cell line A549 under the action of doxorubicin at different concentrations. Methods: After adriamycin treatment on A549 cells for 24 hours, the cell viability was detected by MTT assay. The expression of survivin mRNA and protein was detected by real-time RT-PCR and Western blotting. The expression of p21 protein was detected by Western blotting , And flow cytometry to detect cell cycle changes after drug effect. The cells were further blocked with hydroxyurea in G1 phase to observe the effect of doxorubicin on the cells. Results: Under the different concentration of doxorubicin (from low to high), the cell survival rate decreased first and then rebounded (P <0.01), and accompanied with the high expression of survivin molecule. At the same time, the cell differentiation More inclined to block in the G1 phase. CONCLUSION: The high concentration of doxorubicin blocks cell cycle in G1 phase and induces the high expression of survivin molecule, which enhances the protective effect on cells. This may be the reason of the reverse increase of cell survival rate.