桔梗皂苷-D诱导Hela细胞凋亡的作用及机制研究  

Study on the effect and mechanism of platycodin-D induction on apoptosis of cervical cancer cell line Hela

作　　者：马琳琳[1] 何慧 张世科 李威[1] MA Linlin;HE Hui;ZHANG Shike;LI Wei(The First Affiliated Hospital of Heilongjiang University of Chinese Medicine, Harbin 150040, Heilongjiang, China;Heilongjiang University of Chinese Medicine, Harbin 150040, Heilongjiang, China)

机构地区：[1]黑龙江中医药大学附属第一医院,黑龙江哈尔滨150040 [2]黑龙江中医药大学,黑龙江哈尔滨150040

出　　处：《现代中西医结合杂志》2020年第22期2429-2436,共8页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：黑龙江省中医药管理局一般项目(ZHY18-060)。

摘　　要：目的研究桔梗皂苷-D对宫颈癌Hela细胞凋亡的诱导作用及其机制。方法将体外培养的Hela细胞系随机分为4组,对照组常规培养,桔梗皂苷-D低、中、高剂量组(7,14,28μmol/L)分别加入相应浓度桔梗皂苷-D进行培养。各组细胞培养24 h后,收集细胞用于后续检测。采用CCK8法检测细胞活力,采用流式细胞仪检测细胞周期及细胞凋亡比率,采用TUNEL染色法观察细胞核的形态变化,采用RT-PCR法检测细胞内Bax、P53、Caspase-3、Bcl-2、Hippo信号通路关键因子(MST、LATS、YAP、TAZ、C-myc、β-Catenin)mRNA表达水平,采用Western blot法检测细胞内Bax、P53、Caspase-3、Bcl-2、Hippo信号通路关键因子蛋白表达水平。结果与对照组比较,各组桔梗皂苷-D细胞活力随桔梗皂苷-D浓度增高而显著下降(P均<0.05)。与对照组比较,桔梗皂苷-D各组G1期细胞比例显著增高(P均<0.05),且G1期细胞占比随桔梗皂苷-D浓度增高而逐渐增高,桔梗皂苷-D中、高剂量组G2期及S期Hela细胞占比显著降低(P均<0.05)。与对照组比较,桔梗皂苷-D各组TUNEL染色阳性细胞核的比率和早期凋亡细胞比例均显著增高(P均<0.05),且随桔梗皂苷-D浓度增高而逐渐增高。与对照组比较,桔梗皂苷-D各组Bax、P53、Caspase-3、Bcl-2的mRNA和蛋白表达水平随桔梗皂苷-D浓度增高而逐渐升高(P均<0.05),YAP、TAZ、C-myc、β-Catenin的mRNA和蛋白表达水平随桔梗皂苷-D浓度增高而逐渐降低(P均<0.05)。结论桔梗皂苷-D能显著抑制Hela细胞增殖,促进其凋亡,其机制可能是通过激活Hippo信号通路,下调该通路下游YAP、TAZ的过度表达,上调促凋亡因子Bax、P53、Caspase-3表达,从而使Hela细胞发生G1期阻滞,促进Hela细胞凋亡。Objective It is to explore the inducing effect and mechanism of platycodin-D(PD)on the apoptosis of Hela cells of cervical cancer.Methods The Hela cell lines cultured in vitro were randomly divided into 4 groups,the control group was routinely cultured,and the PD low,medium,and high-dose groups(7,14,28μmol/L)were added with corresponding concentrations of PD for culture.After the cells in each group were cultured for 24 hours,the cells were collected for subsequent detection.CCK8 method was used to detect cell viability,flow cytometry was used to detect cell cycle and cell apoptosis ratio,TUNEL staining method was used to observe morphological changes of cell nuclei,and RT-PCR method was used to detect the expression levels of intracellular Bax,P53,Caspase-3,and Bcl-2,Hippo signaling pathway key factors(MST,LATS,YAP,TAZ,C-myc,β-Catenin)mRNA,Western blot method was used to detect the expression levels of intracellular Bax,P53,Caspase-3,Bcl-2,Hippo signaling pathway key factor protein.Results Compared with the control group,the cell viability decreased significantly with the increase of PD concentration(all P<0.05).Compared with the control group,the proportion of cells in the G1 phase of PD groups was significantly increased(all P<0.05),and the proportion of cells in the G1 phase gradually increased with the concentration of PD.The proportion of Hela cells in G2 and S phases was significantly reduced in the medium and high dose groups of PD(P<0.05).Compared with the control group,the ratio of TUNEL-stained positive nuclei and the proportion of early apoptotic cells in each group of PD were significantly increased(P<0.05),and gradually increased with the concentration of PD.Compared with the control group,the mRNA and protein expression levels of Bax,P53,Caspase-3,and Bcl-2 gradually increased as the concentration of Platycodin-D increased(P<0.05),the mRNA and protein expression levels of YAP,TAZ,C-myc,β-Catenin gradually decreased with the increase of PPD concentration(all P<0.05).Conclusion PD can significantl

关 键 词：桔梗皂苷-D 宫颈癌 HELA细胞 凋亡 机制 

分 类 号：R361.3[医药卫生—病理学]