耐索拉非尼的人肝癌BEL-7402细胞株的建立及其特征

发布时间：2019-05-10 09:04

【摘要】：目的:建立耐索拉非尼人肝癌BEL-7402的细胞株并研究其相关特征。方法:用CCK-8法检测药物对细胞增殖的抑制作用,Coulter计数法分析细胞倍增时间,流式细胞仪检测细胞周期以及活性氧自由基的水平,免疫印迹法检测蛋白的表达。结果:通过浓度递增的方法,诱导人肝癌BEL-7402细胞出现耐药性,得到了低倍耐药的细胞株。结果发现耐药细胞的增殖速度明显减慢,细胞周期发生改变,G1期细胞明显增多。使用相同浓度的索拉非尼处理细胞,与敏感细胞相比较,耐药细胞的活性氧自由基水平明显减少,凋亡相关蛋白P53表达降低,凋亡标志蛋白PARP-1的切割程度变轻。粉防己碱能逆转其耐药性。结论:成功建立了耐索拉非尼的人肝癌BEL-7402细胞株,为索拉非尼的耐药机制及相关药物研究奠定了良好的基础。
[Abstract]:Objective: to establish a cell line resistant to solafini human hepatocellular carcinoma (BEL-7402) and study its related characteristics. Methods: CCK-8 assay was used to detect the inhibitory effect of drugs on cell proliferation, Coulter counting method was used to analyze cell doubling time, flow cytometry was used to detect the cell cycle and the level of reactive oxygen free radicals (Ros), and the expression of protein was detected by immunoblotting. Results: the drug resistance of human hepatocellular carcinoma BEL-7402 cells was induced by increasing concentration, and the cell line with low drug resistance was obtained. The results showed that the proliferation rate of drug-resistant cells was significantly slowed down, the cell cycle was changed, and the number of G _ 1 phase cells was significantly increased. Compared with the sensitive cells, the level of reactive oxygen species (Ros), the expression of apoptosis-related protein p53 (p53) and the cleavage degree of apoptosis marker protein (PARP-1) in the resistant cells were significantly lower than those in the sensitive cells treated with the same concentration of sorafinib. Tetrandrine can reverse its drug resistance. Conclusion: a human liver cancer cell line BEL-7402 resistant to solafenil has been successfully established, which lays a good foundation for the mechanism of Solafinib resistance and related drug research.
【作者单位】：中国医学科学院北京协和医学院医药生物技术研究所;
【基金】：国家自然科学基金资助项目(31471150)
【分类号】：R96

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