Reproductive Medicine Section, Puren Hospital Affiliated to Wuhan University of Science and Technology, Wuhan City, Hubei Province 430081, China.
Department of Obstetrics and Gynecology, Puren Hospital Affiliated to Wuhan University of Science and Technology, Wuhan City, Hubei Province 430081, China.
Cell Mol Biol (Noisy-le-grand). 2021 Sep 29;67(2):14-19. doi: 10.14715/cmb/2021.67.2.3.
The main purpose of the current research work was to study in vitro anticancer effects of betulin in OVCAR-3 human ovarian carcinoma cells along with examining its effects on cellular apoptosis, cell cycle phase distribution, cell migration and invasion and mTOR/PI3K/AKT signalling pathway. The cell proliferation of OVCAR-3 cells at various doses of the drug was studied by CCK8 cell viability assay. Effects on cell apoptosis were studied by fluorescence microscopy and western blot. Effects on cell cycle were evaluated by flow cytometry and western blot. Transwell assays were used to study effects on cell migration and invasion. The results indicated that betulin led to significant reduction of OVCAR-3 cell viability in a dose-dependent as well as time dependent manner. Betulin also led to reduction in cell colonies. The anticancer effects of betulin were due to the induction of apoptosis which was seen by increased apoptotic cells with yellow and orange fluorescence. Betulin prompted mitochondrial apoptosis which was also associated with alteration in the apoptosis-related protein expression (Bax, Bad and Bcl-2 and Bcl-xL). The molecule also led to G2/M phase cell cycle arrest on OVACR-3 ovarian carcinoma cells. It was also observed that betulin could inhibit the migration and invasion of the ovarian cancer cells in a concentration-dependent manner. Betulin molecule also resulted in blocking of mTOR/PI3K/AKT signalling pathway. In conclusion, this study clearly indicates the anticancer effects of betulin natural product in OVCAR-3 human ovarian cancer cells are mediated via apoptosis induction, G2/M phase cell cycle arrest, cell migration and invasion inhibition and targeting of mTOR/PI3K/AKT signalling pathway.
本研究工作的主要目的是研究白桦脂醇在人卵巢癌细胞 OVCAR-3 中的体外抗癌作用，并研究其对细胞凋亡、细胞周期时相分布、细胞迁移和侵袭以及 mTOR/PI3K/AKT 信号通路的影响。通过 CCK8 细胞活力测定研究 OVCAR-3 细胞在不同药物剂量下的细胞增殖。通过荧光显微镜和蛋白质印迹法研究细胞凋亡的影响。通过流式细胞术和蛋白质印迹法评估细胞周期的影响。通过 Transwell 测定研究细胞迁移和侵袭的影响。结果表明，白桦脂醇在剂量和时间依赖性方式下导致 OVCAR-3 细胞活力显著降低。白桦脂醇还导致细胞集落减少。白桦脂醇的抗癌作用是由于诱导细胞凋亡所致，这可通过黄色和橙色荧光的凋亡细胞增加来观察到。白桦脂醇促使线粒体凋亡，这也与凋亡相关蛋白表达（Bax、Bad 和 Bcl-2 和 Bcl-xL）的改变有关。该分子还导致 OVACR-3 卵巢癌细胞的 G2/M 期细胞周期停滞。还观察到白桦脂醇可以浓度依赖性方式抑制卵巢癌细胞的迁移和侵袭。白桦脂醇分子还导致阻断 mTOR/PI3K/AKT 信号通路。总之，本研究清楚地表明，白桦脂醇天然产物在 OVCAR-3 人卵巢癌细胞中的抗癌作用是通过诱导细胞凋亡、G2/M 期细胞周期停滞、抑制细胞迁移和侵袭以及靶向 mTOR/PI3K/AKT 信号通路来介导的。
