冬凌草甲素对肝癌Bel-7402/Sora细胞增殖、凋亡、细胞周期及PI3K/AKT信号通路的影响  

Effects of Oridonin on Proliferation,Apoptosis,Cell Cycle and PI3K/AKT Signaling Pathway of Bel-7402/Sora Cells

作　　者：史国军 叶兴涛 何国浓 施航 李苗 叶晶琳 SHI Guojun;YE Xingtao;HE Guonong(Ningbo Hospital of Chinese Medicine,Ningbo(315010),China)

机构地区：[1]宁波市中医院,宁波315010

出　　处：《浙江中医药大学学报》2024年第9期1102-1109,共8页Journal of Zhejiang Chinese Medical University

基　　金：宁波市自然科学基金项目(2021J305);第五批全国中医临床优秀人才研修项目(国中医药人教函[2022]1号);第四轮省级重点学科“十二五”中西医结合肿瘤学(甬卫发[2021]96号)。

摘　　要：[目的]探讨冬凌草甲素对人肝癌Bel-7402/Sora细胞增殖、凋亡、细胞周期及磷脂酰肌醇3-激酶/蛋白激酶B(phosphatidylinositol 3-kinase/protein kinase B,PI3K/AKT)信号通路的影响。[方法]采用浓度梯度递增法建立Bel-7402/Sora耐药细胞株,以细胞计数法(cell counting kit-8,CCK-8)检测细胞增殖抑制率并计算细胞耐药逆转指数,流式细胞仪检测细胞凋亡和细胞周期,实时荧光定量聚合酶链式反应(Real-time quantitative polymerase chain reaction,RT-PCR)法检测各组细胞中PI3K、AKT、哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)的mRNA表达水平,免疫印迹法检测各组细胞中PI3K、AKT、mTOR的蛋白表达水平。[结果]与空白对照组比较,终浓度为4、8、16、32、64μmol·L^(-1)的冬凌草甲素对Bel-7402/Sora细胞均有较好的抑制作用(P<0.05,P<0.01),且作用呈现浓度依赖性。冬凌草甲素对Bel-7402/Sora细胞耐药逆转指数为2.024。与空白对照组比较,冬凌草甲素组的早期、晚期和总凋亡率均显著升高(P<0.01,P<0.05);与冬凌草甲素组比较,冬凌草甲素+索拉非尼组的早期总凋亡率均显著升高(P<0.05)。与空白对照组比较,冬凌草甲素组的G2期细胞比例显著增加(P<0.01);与冬凌草甲素组比较,冬凌草甲素+索拉非尼组G,期细胞比例增加,G2期细胞比例降低(P<0.01)。与空白对照组比较,冬凌草甲素组PI3K、AKT、mTOR的mmRNA相对表达量均显著降低(P<0.01);与冬凌草甲素组比较,冬凌草甲素+索拉非尼组PI3K、AKT、mTOR的mRNA相对表达量均显著降低(P<0.01)。与空白对照组比较,冬凌草甲素组PI3K、AKT、蛋白相对表达量均显著降低(P<0.01),mTOR蛋白表达无统计学意义(P>0.05);与冬凌草甲素组比较,冬凌草甲素+索拉非尼组AKT、蛋白表达下降,差异有统计学意义(P<0.01),PI3K、mTOR蛋白差异无统计学意义(P>0.05)。[结论]冬凌草甲素能抑制Bel-7402/Sora细胞增殖,诱导细胞凋亡,阻碍细胞周期[Objective]To study the effects of oridonin on Bel-7402/Sora cell proliferation,apoptosis,cycle and phosphatidylinositol 3-kinase/protein kinase B(PI3K/AKT)signaling pathway.[Methods]The concentration gradient increasing method was used to establish Bel-7402/Sora drug-resistant cell line.Cell counting kit-8(CCK-8)method was used to detect cell proliferation inhibition rate and calculate cell resistance reversal index,flow cytometry was used to detect the cell apoptosis and cell cycle,Real-time quantitative polymerase chain reaction(RT-PCR)method was used to detect mRNA expression level of PI3K,AKT and mammalian target of rapamycin(mTOR),Western blot method was used to detect PI3K,AKT,mTOR protein expression levels.[Results]Compared with blank control group,oridonin with final concentrations of 4,8,16,32,64μmol·L-1 had a better proliferation inhibition on Bel-7402/Sora cells(P<0.05,P<0.01),and the effect was dependent on concentration.The resistance reversal index of oridonin on Bel-7402/Sora cells is 2.024.Compared with blank control group,the early,late and total apoptosis rate of oridonin group were significantly increased(P<0.01,P<0.05);compared with oridonin group,the early and total apoptosis rate of oridonin jointed Sorafenib group were significantly increased(P<0.05).Compared with blank control group,the proportion of G2 phase cells of oridonin group was significantly increased(P<0.01);compared with oridonin group,the proportion of G1 phases cells of oridonin jointed Sorafenib group was significantly increased,and G2 phases cells was significantly decreased(P<0.01).Compared with blank control group,PI3K,AKT and mTOR mRNA expression levels of oridonin group were significantly reduced(P<0.01);compared with oridonin group,PI3K,AKT and mTOR mRNA expression levels of oridonin jointed Sorafenib group were significantly reduced(P<0.01).Compared with blank control group,PI3K and AKT protein expression levels of oridonin group were significantly reduced(P<0.01),and mTOR protein expression levels were no statistic

关 键 词：冬凌草甲素 肝癌 Bel-7402/Sora细胞 细胞凋亡 细胞周期 PI3K/AKT信号通路 

分 类 号：R331[医药卫生—人体生理学]