【摘要】 目的：探讨三叶青乙酸乙酯提取物（SYQY）抑制前列腺癌PC-3细胞增殖的作用机制。方法：将4种提取方式的三叶青予PC-3细胞给药，以CCK-8试验筛选最佳提取方式的药物用于后续试验。SYQY给药，克隆形成试验检测细胞增殖情况。网络药理学进行靶点的获取，并将三叶青主要成分与铁死亡关键蛋白谷胱甘肽过氧化物酶4（GPX4）进行分子对接。试剂盒检测细胞还原型谷胱甘肽（GSH）、丙二醛（MDA）及活性氧（ROS）含量；q RT-PCR及Western Blot分别检测铁死亡相关基因P53、SLC7A11、GPX4 mRNA及蛋白表达。使用P53小干扰RNA转染细胞并同时SYQY给药，进一步检测细胞增殖情况。结果：SYQY可在一定程度上呈浓度及时间依赖性抑制PC-3细胞增殖能力（P<0.01）,48 h时的药物IC₅₀值为3.24μg/mL。三叶青主要成分儿茶素与GPX4残基形成了7个稳定氢键，并具有良好的结合活性。与DMSO组比较，SYQY给药可显著降低GSH含量（P<0.05,P<0.01），升高MDA及ROS水平（P<0.01）;P53 mRNA及蛋白表达水平显著升高（P<0.01），而SLC7A11、GPX4 mRNA和蛋白表达水平显著降低（P<0.01）。P53沉默（si-P53）并同时SYQY给药可部分逆转SYQY单独给药对PC-3细胞的增殖抑制作用（P<0.01,P<0.05）。结论：SYQY可能通过激活P53/SLC7A11/GPX4信号通路促进铁死亡，发挥抑制PC-3细胞的增殖作用。更多还原

【Abstract】 Objective:To investigate the possible mechanism of Tetrastigma hemsleyanum ethyl acetate extract （SYQY）on the proliferation ability of PC-3 cells in prostate cancer.Methods:PC-3 cells were administered by 4 extractions of Tetrastigma hemsleyanum,then the best extraction was selected by CCK-8 trial for subsequent tests.After administration of SYQY,the proliferation ability of PC-3 cells was detected by the clonal formation assay.Network pharmacology was used to dock between the main components of Tetrastigma hemsleyanum and the key protein of ferpptosis glutathione peroxidase 4 （GPX4）.The contents of glutathione （GSH）,malondialdehyde （MDA） and reactive oxygen （ROS） were detected by the corresponding kit.The mRNA and protein levels of P53,SLC7A11 and GPX4 were measured by qRT-PCR and Western Blot.At last,cells were transfected with P53 small interfering RNA and administered with SYQY at the same time,the cells proliferation was measured.Results:SYQY could significantly inhibit PC-3 cells’proliferation in a concentration-dependent and time-dependent manner（P<0.01）.The IC₅₀ of SYQY at 48 h was 3.24μg/mL.The main component of Tetrastigma hemsleyanum,catechin,formed 7stable hydrogen bonds with GPX4’s residues,which with good binding activity.Compared with DMSO,SYQY administration significantly reduced GSH content （P<0.05,P<0.01）,while increasing MDA and ROS levels （P<0.01）;the expression levels of P53 mRNA and protein were significantly increased （P<0.01）,while the expression levels of SLC7A11 and GPX4 were signi?cantly decreased （P<0.01）.Silencing P53 and simultaneous SYQY administration partially reversed the inhibitory effect of SYQY alone on the proliferation of PC-3 cells （P<0.01,P<0.05）.Conclusion:SYQY may inhibit the proliferation and promote ferroptosis of PC-3 cells by activating the P53/SLC7A11/GPX4 signaling pathway.更多还原