【摘要】：目的:建立耐索拉非尼人肝癌BEL-7402的細胞株并研究其相關特征。方法:用CCK-8法檢測藥物對細胞增殖的抑制作用,Coulter計數法分析細胞倍增時間,流式細胞儀檢測細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。結果:通過濃度遞增的方法,誘導人肝癌BEL-7402細胞出現耐藥性,得到了低倍耐藥的細胞株。結果發(fā)現耐藥細胞的增殖速度明顯減慢,細胞周期發(fā)生改變,G1期細胞明顯增多。使用相同濃度的索拉非尼處理細胞,與敏感細胞相比較,耐藥細胞的活性氧自由基水平明顯減少,凋亡相關蛋白P53表達降低,凋亡標志蛋白PARP-1的切割程度變輕。粉防己堿能逆轉其耐藥性。結論:成功建立了耐索拉非尼的人肝癌BEL-7402細胞株,為索拉非尼的耐藥機制及相關藥物研究奠定了良好的基礎。
[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)學科學院北京協和醫(yī)學院醫(yī)藥生物技術研究所;
【基金】：國家自然科學基金資助項目(31471150)
【分類號】：R96

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