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作 者:宋杰[1] 祝丽双[1] 陈佳[1] 陆慧琦[1] 韩焕兴[1]
机构地区:[1]第二军医大学长征医院转化医学中心,上海200433
出 处:《现代生物医学进展》2014年第12期2255-2258,共4页Progress in Modern Biomedicine
基 金:国家自然科学基金面上项目(81172805)
摘 要: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 investigate the protential mechanism of the surviva rate of human lung cancer cell line A549 affected by adriamycin.Methods:Cells viability was detected by MTT after cells were cultured for 24 hours in present of adriamycin.Survivin gene expression was detected by quantitative real-time PCR,and western blotting was used to observe protein expression of Survivin and p21,while cell cycle arrest was detected by flow cytometry analysis.Cells viability changes after cell cycle was blocked in G1 phase were observed by hydroxyurea.Results:After being cultured under different concentrations of adriamycin treatment(from low to high),cell viability decreased first and then increased(P〈0.01),and accompanied with survivin overexpression.Flow cytometry analysis confirmed that cell differentiation was more likely to be arrested at G1 phase under treatment of adriamycin.Conclusion:High concentration of adriamycin arrested cell differation at G1 phase and induced survivin overexpression,which enhanced the protective effect of cells themselves.This may be the reason why cells viability increased instead under high concentration of adriamycin teatment.
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