栀子苷抑制PI3K/AKT/mTOR通路对宫颈癌细胞增殖、侵袭、自噬的影响  

Effects of Geniposide on the Proliferation,Invasion,and Autophagy of Cervical Cancer Cells by Inhibiting the PI3K/AKT/mTOR Pathway

作　　者：田甜 李刚 张峰 徐淑娜 李伟华 TIAN Tian;LI Gang;ZHANG Feng;XU Shuna;LI Weihua(Department of Pharmacy,Jinan People's Hospital,Jinan 271100,China;Department of Science and Education,Jinan People's Hospital,Jinan 271100,China)

机构地区：[1]济南市人民医院,药剂科,济南271100 [2]济南市人民医院,科教科,济南271100

出　　处：《中国细胞生物学学报》2025年第4期822-831,共10页Chinese Journal of Cell Biology

基　　金：济南市科技创新发展计划(批准号:202134050)资助的课题。

摘　　要：该文旨在探究栀子苷(GEN)抑制磷酸肌醇3-激酶(PI3K)/蛋白激酶B(AKT)/哺乳动物雷帕霉素靶标(m TOR)通路对宫颈癌细胞增殖、侵袭、自噬的影响。将He La细胞分别用不同浓度(0、25、50、100μmol/L) GEN处理或用100μmol/L GEN和10μmol/L 740Y-P共同处理, MTT和集落形成实验检测GEN对He La细胞增殖的影响;Transwell小室和细胞划痕实验检测GEN对He La细胞侵袭和迁移的影响;流式细胞术检测GEN对He La细胞凋亡的影响;吖啶橙(AO)染色以测定自噬过程中出现的酸性囊泡细胞器(AVO)数量;LC3免疫荧光检测He La细胞自噬情况;qRT-PCR检测He La细胞PI3K、PTEN、AKT、TSC2和m TOR mRNA表达水平;蛋白质免疫印迹检测He La细胞自噬和PI3K/AKT/m TOR信号通路相关蛋白的表达水平。与GEN-0组比较,不同浓度GEN组HeLa细胞存活率降低,集落形成数、侵袭数减少,迁移率, PI3K、AKT和m TOR mRNA表达水平及p-PI3K/PI3K、p-AKT/AKT和pm TOR/m TOR水平均下降(P<0.05),细胞凋亡率、AVO数量、LC3荧光强度、PTEN mRNA表达水平、TSC2 mRNA表达水平、Beclin-1蛋白表达水平、LC3II/I蛋白表达水平上升(P<0.05);与GEN-100组比较, GEN-100+740Y-P组He La细胞存活率增加,集落形成数、细胞侵袭数增多,迁移率、PI3K、AKT和m TOR mRNA表达水平及p-PI3K/PI3K、p-AKT/AKT和p-m TOR/m TOR水平上升(P<0.05),细胞凋亡率、AVO数量、LC3荧光强度、PTEN mRNA表达水平、TSC2 mRNA表达水平、Beclin-1蛋白表达水平、LC3II/I蛋白表达水平下降(P<0.05)。GEN通过抑制PI3K/AKT/m TOR通路抑制宫颈癌细胞增殖和侵袭,促进细胞自噬。This study aims to investigate the effects of GEN(geniposide)on the proliferation,inva-sion,and autophagy of cervical cancer cells by inhibiting the PI3K(phosphoinositol 3-kinase)/AKT(pro-tein kinase B)/mTOR(mammalian target of rapamycin)pathway.HeLa cells were treated with different concentrations(0,25,50,100μmol/L)of GEN or with a combination of 100μmol/L GEN and 10μmol/L 740Y-P.MTT and colony formation assays were used to detect the effect of GEN on HeLa cell proliferation.Transwell chamber and cell scratch assay were used to detect the effects of GEN on HeLa cell invasion and migration.Flow cytometry was used to detect the effect of GEN on apoptosis of HeLa cells.AO(acridine orange)staining was used to determine the number of AVOs(acidic vesicle organelles)presenting during autophagy.Autophagy of HeLa cells was detected by LC3 immunofluorescence.The mRNA expression levels of PI3K,PTEN,AKT,TSC2 and mTOR in HeLa cells were detected by qRT-PCR.Western blot was used to detect the expression levels of autophagy and PI3K/AKT/mTOR signaling pathway related proteins in HeLa cells.Compared with the GEN-O group,the survival rate,colony formation number,cell invasion number,migration rate,PI3K,AKT and mTOR mRNA expression,p-PI3K/PI3K,p-AKT/AKT,and p-mTOR/mTOR levels of HeLa cells in different concentrations of GEN groups decreased(P<0.05),and the apoptosis rate,AVO quantity,LC3 positive rate,PTEN and TSC2 mRNA expression,Beclin-1 and LC3II/I protein expression increased(P<0.05).Compared with the GEN-100 group,the survival rate and colony formation number,cell invasion number,migration rate,PI3K,AKT and mTOR mRNA expression,p-PI3K/PI3K,p-AKT/AKT,and p-mTOR/mTOR levels of HeLa cells in the GEN-100+740Y-P group increased(P<0.05),and the apoptosis rate,AVO quantity,LC3 positive rate,PTEN and TSC2 mRNA expression,Beclin-1 and LC3II/I protein expression decreased(P<0.05).GEN inhibits the PI3K/AKT/mTOR pathway to suppress the proliferation and invasion of cervical cancer cells and promote autophagy.

关 键 词：栀子苷 宫颈癌细胞 磷酸肌醇3-激酶 哺乳动物雷帕霉素靶标 自噬 

分 类 号：R737.33[医药卫生—肿瘤]