摘要
目的探讨对叶百部碱抗卵巢癌的作用机制。方法以人卵巢癌SKOV3细胞为研究对象,设置对照组、顺铂(15μg/mL)组和对叶百部碱(50、75、100μg/mL)组。采用MTT法检测对叶百部碱对细胞活力的影响;光学显微镜观察药物干预后细胞形态;EdU实验检测对叶百部碱对细胞增殖活性的影响;克隆形成实验检测细胞克隆能力;划痕实验和Transwell实验观察细胞迁移和侵袭能力;Hoechst 33258核染色观察细胞凋亡形态;流式细胞术检测细胞凋亡和周期分布情况;Western blotting检测B细胞淋巴瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl-2关联X蛋白(Bcl-2 associated X protein,Bax)、细胞周期蛋白D1(Cyclin D1)、骨髓细胞瘤病毒癌基因(cellular-myelocytomatosis viral oncogene,c-myc)蛋白、β-连环蛋白(β-catenin)、E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)表达。结果与对照组比较,对叶百部碱对SKOV3细胞具有显著的增殖抑制作用(P<0.01);对叶百部碱能改变细胞形态,细胞出现凋亡迹象;对叶百部碱显著抑制细胞克隆形成及迁移、侵袭能力(P<0.01);对叶百部碱组细胞荧光强度增强,细胞核固缩破裂,细胞凋亡率显著升高(P<0.01),G0/G1期和G2/M期细胞比例显著降低(P<0.01),S期细胞比例显著升高(P<0.01);对叶百部碱组Bax/Bcl-2、E-cadherin蛋白表达显著上调(P<0.01),Cyclin D1、c-myc、β-catenin、N-cadherin蛋白表达显著下调(P<0.05、0.01)。结论对叶百部碱通过下调Wnt/β-catenin信号通路及上调Bax/Bcl-2的表达,促进SKOV3细胞凋亡,抑制其增殖、迁移与侵袭,并将细胞阻滞在S期,进而发挥抗肿瘤作用。
Objective To explore the mechanism of tuberostemonine against ovarian cancer.Methods Human ovarian cancer SKOV3 cells were used as the research object,and control group,cisplatin(15μg/mL)group,tuberostemonine(50,75,100μg/mL)groups were set up.MTT method was used to detect the effect of tuberostemonine on cell viability;Cell morphology after drug intervention was observed using optical microscope;EdU experiment was used to detect the effect of tuberostemonine on cell proliferation activity;Cell cloning ability was detected through clone formation experiment;Scratch and Transwell experiments were used to observe cell migration and invasion abilities;Hoechst 33258 nuclear staining was used to observe the morphology of cell apoptosis;Cell apoptosis and cycle distribution were detected by flow cytometry;Western blotting was used to detect the expressions of B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),Cyclin D1,cellular-myelocytomatosis viral oncogene(c-myc),β-catenin,E-cadherin and N-cadherin.Results Compared with control group,tuberostemonine had a significant inhibitory effect on the proliferation of SKOV3 cells(P<0.01);Tuberostemonine could change cell morphology and cells were shown signs of apoptosis;Tuberostemonine could significantly inhibit cell clone formation,migration and invasion abilities(P<0.01);The fluorescence intensity of cells in tuberostemonine group was enhanced,the cell nucleus was pyknotic and ruptured,and the apoptosis rate was significantly increased(P<0.01),the proportions of G0/G1 and G2/M phase cells were significantly reduced(P<0.01),while the proportion of S phase cells was significantly increased(P<0.01);The expressions of Bax/Bcl-2 and E-cadherin proteins were significantly up-regulated(P<0.01),while the expressions of Cyclin D1,c-myc,β-catenin and N-cadherin proteins were significantly down-regulated(P<0.05,0.01)in tuberostemonine group.Conclusion Tuberostemonine promotes SKOV3 cells apoptosis,inhibits its proliferation,migration and invasion,and blocks cells in the S phase by down-regulating Wnt/β-catenin signaling pathway and up-regulating the expression of Bax/Bcl-2,thereby exerting anti-tumor effect.
作者
何石燕
英菲雨
夏星
高红伟
王孝勋
HE Shiyan;YING Feiyu;XIA Xing;GAO Hongwei;WANG Xiaoxun(Guangxi University of Chinese Medicine,Nanning 530200,China;Guangxi Key Laboratory of Zhuang and Yao Ethnic Medicine,Nanning 530200,China;Collaborative Innovation Center of Zhuang and Yao Ethnic Medicine,Nanning 530200,China)
出处
《中草药》
CAS
CSCD
北大核心
2024年第17期5887-5897,共11页
Chinese Traditional and Herbal Drugs
基金
广西壮瑶药重点实验室(桂科基字[2014]32号)
壮瑶药协同创新中心(桂教科研[2013]20号)
广西一流学科中药学(民族药学)(桂教科研[2018]12号)。
