| 史国军,叶兴涛,何国浓,等.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[J].浙江中医药大学学报,2024,48(9):1102-1109. |
| 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 |
| DOI：10.16466/j.issn1005-5509.2024.09.006 |
| 中文关键词: 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 |
| 英文关键词: oridonin liver cancer Bel-7402/Sora cells cell apoptosis cell cycle PI3K/AKT signaling pathway |
| 基金项目: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) |
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| 中文摘要: |
| [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、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。 |
| 英文摘要: |
| [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. |
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