褪黑素通过下调HIF-1α表达增强卵巢癌细胞OVCAR3顺铂敏感性

Melatonin enhances cisplatin sensitivity in OVCAR3 ovarian cancer cell through down regulation of HIF-1αexpression

目的:探讨褪黑素对卵巢癌细胞顺铂敏感性的影响及其机制。方法:采用不同浓度褪黑素联合顺铂作用于卵巢癌细胞OVCAR3,MTT法检测细胞增殖,流式细胞术检测细胞凋亡,Western blot检测凋亡相关蛋白及信号通路蛋白AKT及ERK的磷酸化,同时检测褪黑素对HIF-1α蛋白表达的影响。结果:MTT结果显示,褪黑素和顺铂联合作用能够显著抑制OVCAR3细胞增殖(P<0.05)。流式细胞仪检测结果显示,褪黑素和顺铂联合作用相较于单独顺铂作用或褪黑素作用能够显著增加OVCAR3细胞的凋亡。Western blot结果显示,褪黑素与顺铂联合作用能够增加促凋亡蛋白BAX及Caspase3的表达,抑制抗凋亡蛋白BCL2的表达,检测信号通路结果显示,褪黑素与顺铂联合作用能够促进AKT及ERK的磷酸化,同时褪黑素能够抑制HIF-1α蛋白的表达。结论:褪黑素通过抑制HIF-1α的表达使OVCAR3细胞对顺铂敏感。

Objective:To investigate the effect of melatonin on cisplatin sensitivity in ovarian cancer cells and its mechanism.Methods:Ovarian cancer cells OVCAR3 were treated with different concentrations of melatonin or combined with cisplatin.Cell proliferation was detected by MTT assay.Cell apoptosis was detected by flow cytometry.The expression of apoptosis related proteins and signal pathway proteins AKT and ERK phosphorylation was detected by Western blot,and the effect of melatonin on HIF-1αexpression was detected.Results:MTT results showed that the combination of melatonin and cisplatin could significantly inhibit the proliferation of OVCAR3 cells(P<0.05).The results of flow cytometry showed that the combined action of melatonin and cisplatin could significantly increase the apoptosis ratio of OVCAR3 cells compared with the action of cisplatin alone or the action of melatonin.Western blot results showed that the combined effect of melatonin and cisplatin could increase the expression of pro-apoptotic protein BAX and Caspase3,and inhibit the expression of anti-apoptotic protein BCL2.The detection of signal pathway related proteins showed that the combined effect of melatonin and cisplatin could promote the phosphorylation of AKT and ERK,while melatonin could inhibit the expression of HIF-1α.Conclusion:Melatonin could enhance the sensitivity of OVCAR3 cells to cisplatin through inhibit the expression of HIF-1α.