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目的:观察苦参碱对人肝细胞癌Hep3B细胞增殖、细胞周期、凋亡率以及细胞内β-catenin转录活性的影响,以探讨苦参碱抗肝癌细胞增殖作用的机制。方法:采用cell counting kit-8(CCK-8)细胞计数法测定细胞增殖活性;流式细胞术进行细胞凋亡率和细胞周期分析;双荧光素酶报告系统检测β-catenin的转录活性。结果:苦参碱(50~800mg/L)浓度依赖性地抑制Hep3B细胞的增殖(P<0.01),作用72h的半数抑制浓度(IC50)为312.53mg/L。阴性对照与400mg/L的苦参碱作用48h后,细胞凋亡率分别为4.39%和21.73%,G0/G1期细胞所占比例分别为57.39%和74.48%,S期细胞所占比例为27.67%和12.94%。50、200、800mg/L苦参碱浓度依赖性地抑制Hep3B细胞内β-catenin的转录活性(P<0.05或P<0.01)。结论:苦参碱通过抑制β-catenin的转录活性诱导Hep3B细胞凋亡和细胞周期阻滞,从而表现出抗肝癌细胞增殖的作用。
OBJECTIVE: To observe the effects of matrine on the proliferation, cell cycle and apoptosis of human hepatocellular carcinoma Hep3B cells and the intracellular β-catenin transcriptional activity to explore the mechanism of matrine on the proliferation of Hep3B cells. Methods: Cell proliferation assay was performed with cell counting kit-8 (CCK-8) cell counting method. Flow cytometry was used to analyze the apoptosis rate and cell cycle. The dual luciferase reporter system was used to detect the transcriptional activity of β-catenin. Results: Matrine (50 ~ 800mg / L) inhibited the proliferation of Hep3B cells in a concentration - dependent manner (P <0.01). The IC50 was 72.53mg / L at 72h. After treated with 400mg / L matrine for 48h, the apoptotic rates were 4.39% and 21.73%, respectively. The percentage of cells in G0 / G1 phase was 57.39% and 74.48%, respectively. The proportion of cells in S phase was 27.67 % And 12.94%. Matrine at 50, 200 and 800 mg / L inhibited the transcriptional activity of β-catenin in Hep3B cells in a concentration-dependent manner (P <0.05 or P <0.01). Conclusion: Matrine can induce Hep3B cell apoptosis and cell cycle arrest by inhibiting the transcriptional activity of β-catenin, and thus show the effect of inhibiting the proliferation of Hep3B cells.