三叶青乙酸乙酯提取物通过P53/SLC7A11/GPX4信号通路促进铁死亡抑制前列腺癌PC-3细胞增殖  

Tetrastigma hemsleyanum ethyl acetate extract inhibiting the proliferation of PC-3 cells in prostate cancer by promoting ferroptosis via the P53/SLC7A11/GPX4 signaling pathway

作　　者：游赣花 朱建国[3] 杨萌 文周 李凯 刘旺培 严波[3] YOU Ganhua;ZHU Jianguo;YANG Meng;WEN Zhou;LI Kai;LIU Wangpei;YAN Bo(The Second People's Hospital of Guizhou Province,Guiyang 550004,China;Guizhou Institute for Food and Drug Control,Guiyang 550004,China;Guizhou Province People's Hospital,Guiyang 550002,China;Zunyi Medical University,Zunyi 563000,China;Guiyang University,Guiyang 550005,China;Shaoguan Institute for Food and Drug Control,Shaoguan 521028,China)

机构地区：[1]贵州省第二人民医院,贵阳550004 [2]贵州省食品药品检验所,贵阳550004 [3]贵州省人民医院,贵阳550002 [4]遵义医科大学,遵义563000 [5]贵阳学院,贵阳550005 [6]韶关市食品药品检验所,韶关521028

出　　处：《中华中医药杂志》2024年第9期4688-4693,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.82160551);贵州省高层次创新型人才(No.黔科合平台人才[2018]5639,No.gzwjrs2023-011);贵州省中医药管理局项目(No.QZYY-2021-168);贵州省科技计划项目(No.黔科合基础-ZK[2023]一般210)。

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。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 ICso of SYQY at 48 h was 3.24μg/mL.The main component of Tetrastigma hemsleyanum,catechin,formed 7 stable 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 SLC7A1l and GPX4 were significantly 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.

关 键 词：三叶青 三叶青乙酸乙酯提取物 前列腺癌 PC-3细胞 铁死亡 增殖 

分 类 号：R285[医药卫生—中药学]