竹节香附素A通过PI3K/Akt/mTOR信号通路对人宫颈癌HeLa细胞增殖和凋亡的影响  

Effect of Raddeanin A on the proliferation and apoptosis of cervical cancer HeLa cells through the PI3K/Akt/mTOR signaling pathway

作　　者：张娟[1] 李雪[1] 蒋浩 黄晓萍[1] Zhang Juan;Li Xue;Jiang Hao;Huang Xiaoping(Department of Obstetrics and Gynecology,Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,China)

机构地区：[1]川北医学院附属医院妇产科,南充637000

出　　处：《新医学》2022年第4期266-272,共7页Journal of New Medicine

基　　金：四川省医学科研课题计划(S15026)。

摘　　要：目的 观察竹节香附素A(RDA)对人宫颈癌HeLa细胞增殖和凋亡的影响,并探讨其作用机制。方法分别使用0、5、10、20及40μmol/L的RDA干预HeLa细胞48 h,CCK-8实验测定细胞增殖率,计算半抑制浓度(IC50),确定药物浓度。流式细胞术检测细胞凋亡,蛋白免疫印迹法检测细胞中B细胞淋巴瘤2家族蛋白(Bcl-2)、Bcl-2相关X蛋白(Bax)、活化胱天蛋白酶-3(Cleaved-caspase-3)、磷酸化磷脂酰肌醇3激酶(p-PI3K)、磷酸化蛋白激酶B(p-Akt)、磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)的蛋白表达水平。将HeLa细胞分为4组,即空白对照组、胰岛素样生长因子-1(IGF-1)组、RDA组和联合用药组,检测及比较各组细胞增殖率、凋亡率和p-PI3K、p-Akt和p-mTOR蛋白相对表达量。结果 随着RDA浓度的增加,HeLa细胞的增殖率降低、凋亡率升高(P均<0.05),Bax、Cleaved-caspase-3蛋白相对表达量升高(P均<0.05),Bcl-2、p-PI3K、p-Akt、p-mTOR蛋白相对表达量降低(P均<0.05);IGF-1激活PI3K/Akt/mTOR信号通路后,RDA仍然能抑制HeLa细胞增殖(P均<0.05),降低p-PI3K、p-Akt、p-mTOR蛋白相对表达量(P均<0.05)。结论 RDA可能通过抑制PI3K/Akt/mTOR信号通路对人宫颈癌HeLa细胞产生抑制增殖与促进凋亡的效果。Objective To investigate the effect of Raddeanin A(RDA) on the proliferation and apoptosis of cervical cancer HeLa cells and to explore the possible mechanism. Methods HeLa cells were cultured in vitro and treated with RDA at a concentration of 0, 5, 10, 20, and 40 μmol/L for 48 h. Cell proliferation rate was determined by CCK-8. The half-maximal inhibitory concentration(IC50) was calculated to determine the drug concentration. Flow cytometry was used to detect cell apoptosis, and western blot was employed to detect the expression levels of Bcl-2, Bax, Cleaved-caspase-3, p-PI3K, p-Akt, and p-mTOR proteins. Hela cells were divided into four groups: control group, IGF-1 group, RDA group and combined treatment group. The proliferation rate, apoptosis rate, relative expression levels of p-PI3K, p-Akt and p-mTOR proteins were detected and compared among different groups. Results The proliferation rate of HeLa cells was significantly declined, whereas the apoptosis rate was significantly elevated with the increasing concentration of RDA(both P < 0.05), the relative expression levels of Bax and Cleavedcaspase-3 proteins were significantly up-regulated(both P < 0.05), whereas those of Bcl-2, p-PI3K, p-Akt and p-mTOR proteins were significantly down-regulated(all P < 0.05). After the activation of PI3K/Akt/mTOR signaling pathway by IGF-1, RDA could still inhibit the proliferation of HeLa cells and down-regulate the relative expression levels of p-PI3K, p-Akt and p-m TOR proteins(all P < 0.05). Conclusion RDA can inhibit the proliferation and promote the apoptosis of cervical cancer HeLa cells by suppressing the PI3K/Akt/mTOR signaling pathway.

关 键 词：竹节香附素A HELA细胞 宫颈癌 细胞凋亡 PI3K/Akt/mTOR信号通路 

分 类 号：R285[医药卫生—中药学]