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目的探讨桦褐孔菌醇(inotodiol)对人宫颈癌HeLa细胞诱导凋亡和细胞周期分布的影响及其可能机制。方法不同浓度桦褐孔菌醇与人宫颈癌HeLa细胞共同培养后,采用流式细胞技术检测HeLa细胞周期分布及凋亡率的变化;通过免疫细胞化学法检测cyclinE、p27及bax、bcl-2表达的变化。结果桦褐孔菌醇作用人宫颈癌HeLa细胞后,流式细胞检测显示SubG1峰(细胞凋亡)出现,随桦褐孔菌醇浓度增加和处理时间延长,细胞凋亡率明显增高,且呈时间依赖性关系(r=1.0,P<0.01),S期和G2/M期细胞数显著下降。免疫细胞化学结果显示,cyclinE、bcl-2表达显著降低,p27、bax表达显著增加,与对照组相比均有统计学差异(P<0.05)。结论桦褐孔菌提取物桦褐孔菌醇通过诱导细胞凋亡和G1期阻滞发挥对宫颈癌HeLa细胞的抑制作用,其机制可能与上调bax和下调bcl-2蛋白表达促进细胞凋亡,下调cyclinE和上调负调控因子p27的表达影响细胞周期有关,此结果进一步表明了桦褐孔菌在宫颈癌临床治疗方面的潜在价值。
Objective To investigate the effects of inotodiol on apoptosis and cell cycle distribution of human cervical carcinoma HeLa cells and its possible mechanism. Methods After different concentrations of Inoculation with human cervical carcinoma HeLa cells were inoculated, the cell cycle distribution and apoptosis rate of HeLa cells were detected by flow cytometry. The expressions of cyclinE, p27, bax and bcl-2 were detected by immunocytochemistry Changes in expression. Results The results of flow cytometry showed that sub-G1 peak (apoptosis) appeared after inoculation of Inonellatigma on human cervical cancer HeLa cells. The apoptosis rate of SubG1 peak increased with the increase of inoculum concentration and prolonged treatment time, Time dependent relationship (r = 1.0, P <0.01), S phase and G2 / M phase cells decreased significantly. The results of immunocytochemistry showed that the expressions of cyclinE and bcl-2 were significantly decreased and the expressions of p27 and bax were significantly increased compared with the control group (P <0.05). CONCLUSION: Inonotus iliacus extract inhibits cervical cancer HeLa cells through induction of apoptosis and G1 phase arrest. The mechanism may be related to up-regulation of bax and down-regulation of bcl-2 protein expression, Downregulation of cyclinE and up-regulation of negative regulator p27 affect the cell cycle, the results further demonstrate the potential value of Inonotus obliquus in the clinical treatment of cervical cancer.