摘要
目的探究青藤碱通过抑制PI3K/AKT/mTOR通路诱导自噬相关的卵巢癌细胞凋亡的机制.方法体外培养卵巢癌细胞,采用CCK8检测SIN对其的毒性作用;使用0、0.5、1、3 mmol/L的青藤碱作用A121细胞,检测青藤碱对A121细胞增殖、自噬及凋亡和MPK/mTOR/ULK1通路情况,并检测相关蛋白表达情况.结果青藤碱对卵巢癌细胞毒性明显大于输卵管上皮细胞;相比不使用SIN处理组,使用1和3 mmol/L的青藤碱处理组卵巢癌细胞Ki67蛋白、PCNA蛋白、Bcl-2蛋白、细胞克隆形成率、p-Mtor/mTOR比例均降低(P<0.05),Bax蛋白、Cleaved cas3蛋白、CleavedPARA蛋白、LC3Ⅱ/LC3Ⅰ蛋白、细胞凋亡率、每个细胞中LC3+荧光数目、p-AMPK/AMPK比例、p-ULK1/ULK1比例均升高(P<0.05);与不使用SIN组比较,使用自噬抑制剂Spautin-1处理组Cleaced cas 3蛋白、Cleave PARP蛋白、Bax蛋白表达降低,Bcl-2蛋白表达升高(P<0.05).结论青藤碱可以通过抑制PI3K/AKT/mTOR通路调控卵巢癌细胞的自噬实现抑制其增殖并促进其凋亡.
Objective To explore the mechanism of sinomenine(SIN)inducing apoptosis of autophagy-related ovarian cancer cells by inhibiting PI3/AKT/mTOR pathway.Methods Ovarian cancer cells were cultured in vitro.The toxic effects of SIN on them were detected by CCK8.A121 cells were treated with SIN at 0,0.5,1 and 3 mmol/L,respectively.The effects of SIN on the proliferation,autophagy and apoptosis of A121 cells,and the MPK/mTOR/ULK1 pathway were detected.The expression of related proteins was detected.Results The toxic effects of SIN on ovarian cancer cells were signifcantly greater than on fallopian tube epithelial cells.The Ki67,PCNA and Bel-2 proteins,clone formation rate and p-Mtor/mTOR were significantly decreased,while Bax,cleaved caspase 3,cleaved PARA and LC3 II/LC3 I proteins,apoptosis rate,the number of LC+fluorescence in each cell,p-AMPK/AMPK and p-ULK1/ULK1 were all increased in the group treated with 1 and 3 mmol/L SIN as compared with the group without SIN treatment,(P<0.05 for all).The expression of cleaved caspase 3,cleave PARP and Bax proteins was significantly decreased,while the expression of Bcl-2 protein was significantly increased in the group treated with the autophagy inhibitor Spautin-1 when compared with the group without SIN treatment(P<0.05)Conclusion SIN can regulate the autophagy of ovarian cancer cells by inhibiting PI3 K/AKT/mTOR pathway to inhibit proliferation and promote apoptosis of ovarian cancer cells.
作者
张彩萍
葛俊丽
柏璐
ZHANG Caiping;GE Junli;BAI Lu(Department of Obstetrics and Gynecology,The First Affiliated Hospital of Air Force Medical University,Xi'an,710004,China)
出处
《医学分子生物学杂志》
CAS
2020年第5期359-365,共7页
Journal of Medical Molecular Biology
基金
陕西省自然科学基础研究计划(No.2017JQ8290)。
关键词
卵巢癌
青藤碱
自噬
细胞凋亡
ovarian cancer
sinomenine
autophagy
apoptosis
