牛蒡子苷通过ROS/PI3K/Akt/mTOR通路调控宫颈癌细胞的研究  

Study on arctiin regulates the proliferation and autophagy of cervical cancer cells through ROS/PI3K/Akt/mTOR pathway

作　　者：杨卫[1] 李媛媛[1] 张丽柯 丁岩 YANG Wei;LI Yuanyuan;ZHANG Like;DING Yan(Medical Key Laboratory of Molecular Biology of Tumor in He'nan Province,Nanyang First People's Hospital,Nanyang 473001,China;Department of Radiation Oncology,Nanyang First People's Hospital,Nanyang 473000,China)

机构地区：[1]南阳市第一人民医院河南省肿瘤分子生物学医学重点实验室,南阳473001 [2]南阳市第一人民医院放射肿瘤科,南阳473000

出　　处：《西北药学杂志》2023年第6期89-93,共5页Northwest Pharmaceutical Journal

基　　金：2021年度江苏省放射医学与防护重点实验室开放课题(编号:KJS2105)。

摘　　要：目的探讨牛蒡子苷(arctiin,ARC)对人宫颈癌细胞恶性增殖的抑制作用及其可能的作用机制。方法将不同浓度ARC作用于人宫颈癌细胞Hela,用CCK-8法检测细胞存活情况,筛选最佳抑制浓度;将对数生长期Hela细胞随机分为对照组、ARC组、ARC联合自噬抑制剂(3-MA)组。用CCK-8法检测细胞增殖抑制率;用DCFH-DA检测各组细胞活性氧(reactive oxygen species,ROS)水平;用透射电镜观察各组细胞自噬小体;反转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)和蛋白质印迹法(Western blotting)检测细胞磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K)、蛋白激酶B(protein kinase B,Akt)、哺乳动物雷帕霉菌靶蛋白(mammalian target of rapamycin,mTOR)mRNA和蛋白表达及哺乳动物ATG6同源蛋白(mammalian ATG6 homologous protein,Beclin-1)和微管相关蛋白1轻链3(microtubule-associated protein light chain 3,LC3)蛋白的表达情况。结果与对照组比较,ARC组细胞ROS水平、PI3K、Akt和mTOR mRNA、PI3K、p-Akt以及p-mTOR蛋白的表达水平降低,且Beclin-1和LC3-Ⅱ蛋白表达水平升高(P<0.05)。3-MA可逆转ARC组上述指标(P<0.05)。结论ARC对人宫颈癌细胞Hela的恶性增殖有一定的抑制作用,并能提高宫颈癌细胞自噬水平,可能是通过调节细胞ROS水平,影响ROS/PI3K/Akt/mTOR信号通路发挥作用。Objective To investigate the inhibitory effect of arctiin(ARC)on the malignant proliferation of human cervical cancer cells and its possible mechanism.Methods Different concentrations of ARC were used to act on HeLa cells,and the survival rate of HeLa cells was detected by CCK-8 method.HeLa cells in logarithmic growth phase were randomly divided into control group,ARC group and ARC combined with autophagy inhibitor(3-MA)group.CCK-8 assay was used to detect the inhibition rate of cell proliferation and DCFH-DA was used to detect the level of reactive oxygen species(ROS)in each group.Autophagy bodies were observed by transmission electron microscope.reverse transcription-polymerase chain reaction(RT-PCR)and Western blotting were used to detect the mRNA and protein expression of phosphatidylinositol 3-kinase(PI3K),protein kinase B(Akt),mammalian target protein(mTOR),mammalian ATG6 homologous protein(Beclin-1)and microtubule associated protein 1 light chain 3(LC3).Results Compared with the control group,the levels of ROS,PI3K,Akt and mTOR mRNA expression,PI3K,p-Akt and p-mTOR protein expression were decreased,and the levels of Beclin-1 and LC3-Ⅱprotein were increased in ARC group(P<0.05).3-MA reversed the above indices in the ARC group(P<0.05).Conclusion ARC can inhibit the malignant proliferation of HeLa cells and increase the autophagy of HeLa cells,which may play a role by regulating the level of ROS and affecting the ROS/PI3K/Akt/mTOR signal pathway.

关 键 词：牛蒡子苷 宫颈癌细胞 自噬 磷脂酰肌醇3-激酶 蛋白激酶B 哺乳动物雷帕霉菌靶蛋白 

分 类 号：R965[医药卫生—药理学]