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（15 μg/mL）组和对叶百部碱（50、75、100 μg/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 (15 μg/mL) group, tuberostemonine (50, 75, 100 μg/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.

R285.5

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）