STAT3抑制剂及烟酰胺联合用药对肝癌HepG2细胞增殖的抑制作用及机制研究  

Effects and mechanism of STAT3 inhibitor and nicotinamide combination on the inhibiting proliferation of hepatocarcinoma HepG2 cells

作　　者：詹雯静 梁铭杰 刘媛 黄远凤 王玮璇 ZHAN Wenjing;LIANG Mingjie;LIU Yuan;HUANG Yuanfeng;WANG Weixuan(Institute of Chinese Medicine,Guangdong Pharmaceutical University,Guangzhou 510006,China)

机构地区：[1]广东药科大学中医药研究院,510006

出　　处：《天津医药》2022年第7期686-692,共7页Tianjin Medical Journal

基　　金：广东省基础与应用基础研究基金项目(2019A1515110123,2021A1515012553);广东省医学科学技术研究基金项目(A2019531)。

摘　　要：目的探究STAT3抑制剂(Stattic和S3I-201)以及烟酰胺腺嘌呤二核苷酸(NAD)前体烟酰胺联合用药对肝癌HepG_(2)细胞增殖的影响及机制。方法以肝癌细胞株HepG_(2)为模型,使用STAT3抑制剂、烟酰胺以及两者联合处理细胞,通过细胞计数、Western blot和qPCR等检测药物对细胞增殖、STAT3表达、STAT3下游靶基因[锌指转录因子1(SNAIL1)、血管内皮生长因子A(VEGFA)、锌指结合蛋白1(ZEB1)]表达和与细胞增殖[微小染色体维持蛋白7(MCM7)、增殖标志物Ki-67(MKI67)、Myc原癌基因蛋白(MYC)]、细胞凋亡[B淋巴细胞瘤2相关X蛋白(BAX)、B淋巴细胞瘤-2(BCL-2)、髓样细胞白血病蛋白1(MCL-1)]、上皮-间充质转化[紧密连接蛋白-1(TJP1)、母亲DPP同源物3(SMAD3)]以及糖代谢[己糖激酶2(HK2)、磷酸果糖激酶(PFKL)、M型丙酮酸激酶(PKM)、葡萄糖转运蛋白1(GLUT_(1))、乳酸脱氢酶A(LDHA)]相关基因的mRNA表达水平的影响。结果STAT3抑制剂和烟酰胺处理均能降低STAT3磷酸化水平,抑制细胞增殖,降低细胞增殖、抗凋亡、上皮-间充质转化以及糖代谢相关基因表达水平,上调促凋亡相关基因表达水平。并且,联合用药对STAT3磷酸化水平、细胞增殖以及上述生物过程相关基因的表达水平影响更显著。结论STAT3抑制剂和烟酰胺可抑制肝癌细胞增殖,并且联合用药效果更显著。这种抑制作用可能与促进凋亡和抑制上皮-间充质转化及糖酵解有关。Objective To investigate the effects and underlying mechanisms of the combination of STAT3 inhibitors(Stattic and S3I-201)and nicotinamide adenine dinucleotide(NAD)precursor nicotinamide on the proliferation of hepatocarcinoma HepG_(2) cells.Methods Hepatocarcinoma HepG_(2) cells were treated with STAT3 inhibitor,nicotinamide and their combination.Cell counting,Western blot assay and qPCR were used to examine the effects of drugs on cell proliferation,STAT3 expression and mRNA expression levels of STAT3 downstream target genes(SNAIL1,VEGFA and ZEB1),cell proliferation(MCM7,MKI67 and MYC),apoptosis(BCL-2,MCL-1 and BAX),epithelial-mesenchymal transformation(TJP1 and SMAD3),and glucose metabolism(HK2,PFKL,PKM,GLUT_(1) and LDHA)-related genes.Results Both STAT3 inhibitor and nicotinamide treatment can decrease the phosphorylation level of STAT3,inhibit cell proliferation,downregulate the expression levels of genes related to cell proliferation,anti-apoptosis,EMT,and glucose metabolism,and upregulate the expression levels of genes related to pro-apoptosis.In addition,the combination of drugs showed a more significant effect on the phosphorylation level of STAT3,cell proliferation and expression levels of the above-mentioned cellular process-related genes.Conclusion STAT3 inhibitor and nicotinamide can inhibit the proliferation of hepatocarcinoma cells,and the combined effect is more significant.This inhibition may be related to the promotion of apoptosis,the inhibition of EMT and glycolysis.

关 键 词：癌 肝细胞 HEPG2细胞 STAT3转录因子 烟酰胺 细胞增殖 细胞凋亡 上皮-间质转化 糖酵解 

分 类 号：R735.7[医药卫生—肿瘤]