首页|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

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

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[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、64 μmol·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G1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mRNA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,阻碍细胞周期进程,其机制可能与干预PI3K/AKT信号通路有关.
Effects of Oridonin on Proliferation,Apoptosis,Cell Cycle and PI3K/AKT Signaling Pathway of Bel-7402/Sora Cells
[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 4,8,16,32,64 μmol·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 statistical significance(P>0.05);compared with oridonin group,AKT protein expression levels of oridonin jointed Sorafenib group were significantly reduced(P<0.01),PI3K,mTOR protein expression levels were no statistical significance(P>0.05).[Conclusion]Oridonin can inhibit cell proliferation of Bel-7402/Sora cell,induce apoptosis,hinder cycle progression and its anti-tumor mechanism may be related to intervene in the PI3K/AKT signaling pathway.

oridoninliver cancerBel-7402/Sora cellscell apoptosiscell cyclePI3K/AKT signaling pathway

史国军、叶兴涛、何国浓、施航、李苗、叶晶琳

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宁波市中医院 宁波 315010

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

2024

浙江中医药大学学报
浙江中医药大学

浙江中医药大学学报

CSTPCD
影响因子:1.049
ISSN:1005-5509
年,卷(期):2024.48(9)