硫氧还蛋白还原酶抑制剂乙烷硒啉逆转顺铂耐药及其机制研究（英文）
发布时间：2019-06-10 23:32
【摘要】：目的:研究凋亡调控相关蛋白来了解顺铂耐药成因,同时考察乙烷硒啉(Ethaselen)在K562耐药细胞中逆转顺铂耐药的作用,并初步探讨其作用机制。创新点:首次研究乙烷硒啉在逆转顺铂耐药中的作用,且此作用与乙烷硒啉诱导细胞凋亡相关。方法:通过长时间脉冲诱导得到顺铂耐药K562细胞,并观察耐药细胞形态及倍增时间。采用MTT法考察乙烷硒啉、顺铂及其联用组在不同细胞株间的生长抑制作用。流式细胞术分析细胞凋亡情况以及细胞内活性氧(ROS)水平。最后,通过蛋白质免疫印迹(Western blot)考察凋亡调控相关蛋白水平的变化。结论:脉冲诱导得到的K562耐药细胞对顺铂的耐受性是原K562细胞的5.34倍。形态学观察发现,耐药细胞体积增大,粘附性进一步降低。乙烷硒啉与顺铂联用表现出协同效应。当加入少量的乙烷硒啉(顺铂与乙烷硒啉的摩尔比率为10:1),顺铂作用K562耐药细胞的半抑制浓度(IC50)值可以减少21倍。流式细胞术及Western blot表明,乙烷硒啉能够诱导耐药细胞凋亡。其逆转顺铂耐药主要是通过调控Bcl-2及Bax蛋白比例以及通过提高细胞内活性氧水平引起线粒体通透转运孔道(PTP)蛋白孔道的形成来促使释放细胞色素c,进而引起Caspase凋亡途径。
[Abstract]:Aim: to study the apoptosis regulation related proteins to understand the cause of platinum resistance, and to investigate the reversal effect of ethane selenite (Ethaselen) on platinum resistance in K562 resistant cells, and to explore its mechanism. Innovation: the role of ethane selenite in reversal of platinum resistance was studied for the first time, and this effect was related to apoptosis induced by ethane selenite. Methods: DDP resistant K562 cells were induced by long pulse, and the morphology and doubling time of drug resistant cells were observed. MTT assay was used to investigate the growth inhibition of ethane selenite, DDP and their combination groups among different cell lines. Apoptosis and intracellular reactive oxygen species (Ros) (ROS) levels were analyzed by flow cytometry. Finally, the changes of apoptosis-regulated protein levels were investigated by Western imprinting (Western blot). Conclusion: the drug resistance of K562 cells induced by pulse is 5.34 times as much as that of original K562 cells. Morphological observation showed that the volume of drug-resistant cells increased and the adhesion decreased further. The combination of ethane selenite and DDP showed synergistic effect. When a small amount of ethane selenite was added (the molar ratio of DDP to ethane selenite was 10:1), the semi-inhibitory concentration (IC50) of K562 cells treated with DDP could be reduced by 21 times. Flow cytometry and Western blot showed that ethane selenite could induce apoptosis of drug-resistant cells. The reversal of DDP resistance is mainly due to the regulation of the proportion of Bcl-2 and Bax proteins and the formation of (PTP) protein pores in mitochondrial permeable transport channels by increasing the level of reactive oxygen species (Ros) in the cells to promote the release of cytochrome C. And then the apoptosis pathway of Caspase was induced.
【作者单位】： State
【基金】：supported in part by the National Natural Science Foundation of China(No.81372266) the National Science and Technology Major Project of the Ministry of Science and Technology of China(No.2011zx09101-001-03)
【分类号】：R96
[Abstract]:Aim: to study the apoptosis regulation related proteins to understand the cause of platinum resistance, and to investigate the reversal effect of ethane selenite (Ethaselen) on platinum resistance in K562 resistant cells, and to explore its mechanism. Innovation: the role of ethane selenite in reversal of platinum resistance was studied for the first time, and this effect was related to apoptosis induced by ethane selenite. Methods: DDP resistant K562 cells were induced by long pulse, and the morphology and doubling time of drug resistant cells were observed. MTT assay was used to investigate the growth inhibition of ethane selenite, DDP and their combination groups among different cell lines. Apoptosis and intracellular reactive oxygen species (Ros) (ROS) levels were analyzed by flow cytometry. Finally, the changes of apoptosis-regulated protein levels were investigated by Western imprinting (Western blot). Conclusion: the drug resistance of K562 cells induced by pulse is 5.34 times as much as that of original K562 cells. Morphological observation showed that the volume of drug-resistant cells increased and the adhesion decreased further. The combination of ethane selenite and DDP showed synergistic effect. When a small amount of ethane selenite was added (the molar ratio of DDP to ethane selenite was 10:1), the semi-inhibitory concentration (IC50) of K562 cells treated with DDP could be reduced by 21 times. Flow cytometry and Western blot showed that ethane selenite could induce apoptosis of drug-resistant cells. The reversal of DDP resistance is mainly due to the regulation of the proportion of Bcl-2 and Bax proteins and the formation of (PTP) protein pores in mitochondrial permeable transport channels by increasing the level of reactive oxygen species (Ros) in the cells to promote the release of cytochrome C. And then the apoptosis pathway of Caspase was induced.
【作者单位】： State
【基金】：supported in part by the National Natural Science Foundation of China(No.81372266) the National Science and Technology Major Project of the Ministry of Science and Technology of China(No.2011zx09101-001-03)
【分类号】：R96
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