摘要
目的 探讨紫草素对人卵巢癌HO?8910细胞周期及凋亡诱导的作用及对磷脂酰肌醇3激酶/蛋白激酶B(PI3K/AKT)信号通路的影响.方法 以不同浓度的紫草素干预对数生长期的卵巢癌HO?8910细胞,细胞计数法(CCK?8)法检测细胞增殖抑制率,计算半数抑制浓度(IC50),筛选后续实验紫草素作用浓度.流式细胞仪检测紫草素对HO?8910细胞周期的影响,AnnexinⅤ?FITC/PI双染色法检测紫草素对HO?8910细胞凋亡的影响,蛋白免疫印迹法(Western blotting)检测细胞周期蛋白D1 (Cyclin D1)、Bcl?2相关X蛋白(Bax)、剪切的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase?3)、磷脂酰肌醇?3激酶(PI3K)、蛋白激酶B(AKT)蛋白表达及PI3K和AKT磷酸化水平.结果 不同浓度的紫草素对HO?8910细胞具有不同程度增殖抑制作用,根据IC50筛选出浓度为5和10 μg/ml的紫草素用于后续实验.紫草素能够阻滞细胞周期至G0/G1期,诱导细胞发生凋亡,抑制Cyclin D1蛋白表达,促进Bax和Cleaved Caspase?3蛋白表达,抑制PI3K/AKT信号通路的激活.结论 紫草素能够阻碍卵巢癌HO?8910细胞周期进程,诱导细胞凋亡,其抗肿瘤机制可能与抑制PI3K/AKT信号通路的活化有关.
Objective To investigate the effect of Shikonin on cell cycle and inducted apoptosis of human ovarian cancer HO?8910 cells and its role in the phosphatidylinositol 3?kinase/protein kinase B(PI3K/AKT)signaling pathway.Methods The ovarian cancer HO?8910 cells in the logarithmic growth phase were treated with various doses of Shikonin.CCK?8 assay was used to determine the inhibition rate of cell proliferation.The half?inhibitory concentration(IC50)was calculated and the working concentration of Shikonin was thereby screened for the subsequent experiments.The effect of Shikonin on the HO?8910 cell cycle was measured by flow cytometry.The effect of Shikonin on the apoptosis of HO?8910 cells was detected by Annexin V?FITC/PI double staining.Western blotting was used to examine the protein expression of Cyclin D1,Bcl?2 related X gene(Bax),cleaved cysteine?containing Caspase?3,PI3K,AKT,as well as the PI3K/AKT phosphorylation levels.Results Various doses of Shikonin exhibited different levels of inhibition on HO?8910 cells proliferation.The Shikonin concentrations selected according to IC50 for subsequent experiments were 5μg/ml and 10μg/ml.Shikonin was able to arrest the cell cycle at G0/G1 phase,induce cell apoptosis,inhibit Cyclin D1 protein expression,promote Bax and cleaved caspase?3 protein expression,and inhibit PI3K/AKT signaling pathway activation.Conclusion Shikonin may suppress the cell cycling and induce apoptosis of ovarian cancer HO?8910 cells.Such anti?tumor mechanism may be related to inhibition of PI3K/AKT signaling pathway activation.
作者
袁建华
杨亚运
王蕊
Yuan Jianhua;Yang Yayun;Wang Rui(Department of Obstetrics and Gynecology,Affiliated Heping Hospital of Changzhi Medical College,Changzhi,Shanxi 046000;Shanxi University of Traditional Chinese Medicine,Jinzhong,Shanxi 030024,China)
出处
《中华生物医学工程杂志》
CAS
2019年第2期189-194,共6页
Chinese Journal of Biomedical Engineering
作者简介
通信作者:王蕊,Email:wrhpyy@163.com
