摘要
目的:观察中药白花蛇舌草有效成分2-羟基-3-甲基蒽醌(HMA)对人肝癌HepG2细胞生长的抑制作用,并探讨其可能的作用机制。方法:CCK-8法观察HMA对肝癌SMMC-7721、HepG2细胞活性的影响;Annexin V-FITC/PI双染法结合流式细胞术检测肝癌HepG2细胞凋亡率;RT-PCR法检测凋亡相关基因Bcl-2、Bax、Caspase-9mRNA表达水平;Western Blot法检测STAT3及p-STAT3蛋白质表达水平。结果:25、50、100μmol/L的HMA能显著抑制肝癌SMMC-7721、HepG2细胞的增殖(P<0.05),其中肝癌HepG2细胞对HMA更为敏感。IL-6能促进HepG2细胞的生长(P<0.05),而HMA能够抑制IL-6对HepG2细胞的促增殖作用(P<0.05)。HMA能下调抗凋亡基因Bcl-2mRNA的表达(P<0.05),上调促凋亡基因Bax和Caspase-9 mRNA的表达(P<0.05),从而促进HepG2细胞凋亡。此外,作用于HepG2细胞48h后,HMA能抑制由IL-6介导的STAT3的磷酸化(P<0.05)。结论:HMA对肝癌HepG-2细胞具有体外抑制及诱导凋亡作用,其机制可能与抑制IL-6/STAT3信号通路有关。
Objective:To observe the inhibitory effects of 2-hydroxyl group-3-tectoquinone on the growth of HepG2 cells in human liver cancer and to explore its possible mechanism.Methods:Effects of 2-hydroxyl group-3-tectoquinone on the liver cancer cell activities of SMMC-7721 and HepG2 were detected by CCK-8 method;HepG2 ceils apoptotic rate were detected by Annexin V-FITC/PI double staining combined with flow cytometry.The mRNA expression levels of Bcl-2,Bax and Caspase-9 were detected by RT-PCR;The protein expression levels of STAT3 and p-STAT3 were detected by Western Blot.Results: 2-hydroxyl group-3-tectoquinone (25,50,100μmol/L)could significantly inhibited the proliferation of liver cancer SMMC-7721 cells and HepG2 cells (P<0.05).HepG2 cells were more sensitive to 2-hydroxyl group-3-tectoquinone.IL-6 promoted the growth of HepG2 cells (P<0.05),while 2-hydroxyl group-3-tectoquinone could inhibit the promoting proliferation effect induced by IL-6 (P<0.05).2-hydroxyl group-3-tectoquinone could downregulated the mRNA expression level of Bcl-2 and could upregulated the expression levels of Bax and Caspase-9(P<0.05).Thus,the apoptosis of HepG2 ceils were promoted.In addition,after 2-hydroxyl group-3-tectoquinone having an effect on HepG2 cells for 48h,it could inhibit the phosphorylation of STAT3 mediated by IL-6 (P<0.05).Conclusion:2-hydroxyl group-3-tectoquinone has effects of inhibiting and inducing apoptosis in vitro on liver cancer HepG2 cells,and its mechanism may be related to inhibition of IL-6/STAT3 signaling pathway.
作者
孙超
吴铭杰
江泽群
沈卫星
程海波
李黎
李文婷
杨婧
陈婷婷
饶希午
吴勉华
SUN Chao;WU Ming-jie;JIANG Ze-qun;SHEN Wei-xing;CHENG Hai-bo;LI Li;LI Wen-ting;YANG Jing;CHEN Ting-ting;RAO Xi-wu;WU Mian-hua(Nanjing University of Chinese Medicine,Jiangsu Collaborative Innovation Center of Chinese Medicine Prevention and Treatment of Tumor,Nanjing 210023,China;Translational Medicine Research Center of Nanjing University of Chinese Medicine,Nanjing 210023,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2018年第12期5346-5350,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金面上项目(No.81273717)
国家自然科学基金青年科学基金项目(No.81503535).