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目的:探讨桦褐孔菌醇对人卵巢癌SKOV3细胞周期的影响及其可能机制。方法:应用不同浓度桦褐孔菌醇与人卵巢癌SKOV3细胞共同培养48 h后,采用MTT法观察桦褐孔菌醇对人卵巢癌SKOV3的增殖抑制作用;流式细胞技术检测桦褐孔菌醇作用后SKOV3细胞周期及凋亡率的变化;免疫细胞化学法检测桦褐孔菌醇处理后SKOV3细胞CyclinE,P27蛋白表达变化。结果:桦褐孔菌醇对SKOV3细胞的体外增殖可产生明显的抑制作用(P<0.01),该作用能够随桦褐孔菌醇浓度的增高而增高;与对照组比较,桦褐孔菌醇作用人卵巢癌SKOV3细胞48h后,S期细胞减少,而G0/G1期细胞逐渐增多,且subG1期细胞也显著增高(P<0.01);桦褐孔菌醇引起CyclinE蛋白水平的表达降低,P27蛋白水平表达增加(P<0.01)。结论:桦褐孔菌醇通过下调CyclinE和上调细胞周期负调控因子P27的表达,影响细胞周期,致使产生G1期阻滞,从而抑制细胞增殖和诱导细胞凋亡。
OBJECTIVE: To investigate the effect of inonitiao on the cell cycle of human ovarian cancer cell line SKOV3 and its possible mechanism. Methods: After incubation with different concentrations of Initrobacillin and human ovarian cancer SKOV3 cells for 48 h, MTT assay was used to observe the inhibition of proliferation of human ovarian cancer cell line SKOV3 by flow cytometry. The changes of cell cycle and apoptosis rate of SKOV3 cells after alcohol treatment were observed. The expressions of CyclinE and P27 protein in SKOV3 cells were detected by immunocytochemistry. Results: Initiaool could significantly inhibit the proliferation of SKOV3 cells in vitro (P <0.01), and this effect could be increased with the increase of the concentration of Inonotoxin. Compared with the control group, After treated with SKOV3 cells for 48h, the number of S phase cells decreased and the number of cells in G0 / G1 phase increased gradually, and the number of subG1 phase cells increased significantly (P <0.01). The expression of CyclinE protein was decreased in inoculation of INOVA, P27 Protein expression increased (P <0.01). CONCLUSION: Inonotoxin inhibits cell proliferation and induces cell apoptosis by down-regulating CyclinE and up-regulating the expression of P27, a cell cycle-negative regulator of cell cycle, resulting in G1 phase arrest.