【摘要】：目的:建立耐索拉非尼人肝癌BEL-7402的細(xì)胞株并研究其相關(guān)特征。方法:用CCK-8法檢測藥物對細(xì)胞增殖的抑制作用,Coulter計(jì)數(shù)法分析細(xì)胞倍增時(shí)間,流式細(xì)胞儀檢測細(xì)胞周期以及活性氧自由基的水平,免疫印跡法檢測蛋白的表達(dá)。結(jié)果:通過濃度遞增的方法,誘導(dǎo)人肝癌BEL-7402細(xì)胞出現(xiàn)耐藥性,得到了低倍耐藥的細(xì)胞株。結(jié)果發(fā)現(xiàn)耐藥細(xì)胞的增殖速度明顯減慢,細(xì)胞周期發(fā)生改變,G1期細(xì)胞明顯增多。使用相同濃度的索拉非尼處理細(xì)胞,與敏感細(xì)胞相比較,耐藥細(xì)胞的活性氧自由基水平明顯減少,凋亡相關(guān)蛋白P53表達(dá)降低,凋亡標(biāo)志蛋白PARP-1的切割程度變輕。粉防己堿能逆轉(zhuǎn)其耐藥性。結(jié)論:成功建立了耐索拉非尼的人肝癌BEL-7402細(xì)胞株,為索拉非尼的耐藥機(jī)制及相關(guān)藥物研究奠定了良好的基礎(chǔ)。
[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.
【作者單位】： 中國醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院醫(yī)藥生物技術(shù)研究所;
【基金】：國家自然科學(xué)基金資助項(xiàng)目(31471150)
【分類號】：R96

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相關(guān)期刊論文 前4條

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