基于JAK2/STAT3信号通路研究银杏内酯B对食管癌细胞增殖及糖酵解水平的影响  

Effect of ginkgolide B on the proliferation and glycolysis level of esophageal cancer cells based on JAK2/STAT3 signaling pathway

作　　者：殷星[1] 侯永超[1] 杨利姣 YIN Xing;HOU Yong-chao;YANG Li-jiao(Department of oncology,Handan First Hospital,Handan 056002,Hebei,China)

机构地区：[1]邯郸市第一医院肿瘤科,056002

出　　处：《现代消化及介入诊疗》2023年第4期442-446,共5页Modern Interventional Diagnosis and Treatment in Gastroenterology

基　　金：河北省医学科学研究课题(20231909)。

摘　　要：目的探究银杏内酯B(GB)对食管癌细胞增殖、糖酵解的影响及作用机制。方法体外培养人食管癌OE19,分为对照组(Con组,不做干预)、低/中/高剂量实验组(L/M/H组,分别加入12.5、25、50μmol·L^(-1)GB)、顺铂组(CP组,4 mg·L^(-1)CP),筛选GB最适作用浓度进行后续实验。而后分为Con组、GB组(12.5μmol·L^(-1)GB)、CP组、inhibitor组(12.5μmol·L^(-1)GB+10μmol·L^(-1)JAK2/STAT3通路抑制剂AG490)、activator组(12.5μmol·L^(-1)GB+0.5μmol·L^(-1)JAK2/STAT3通路激活剂Colivelin),干预24 h。采用细胞计数试剂盒-8(CCK-8)、5-乙炔基-2′脱氧尿嘧啶核苷(EdU)法、乳酸检测试剂盒、葡萄糖检测试剂盒、实时荧光定量PCR(RT-qPCR)和蛋白免疫印迹(WB)法检测细胞活力、增殖率、乳酸含量、葡萄糖消耗水平、[细胞周期素D1(Cyclin D1)、M2型丙酮酸激酶(PKM2)]mRNA和蛋白、Janus激酶2(JAK2)/转录激活因子3(STAT3)信号通路相关蛋白表达水平。结果与Con组相比,L/M/H组与CP组细胞活力降低(P<0.05),因此本研究选择较低浓度的12.5μmol·L^(-1)GB作为GB组进行后续实验。与Con组相比,GB组和CP组细胞增殖率、乳酸含量、葡萄糖消耗水平、(Cyclin D1、PKM2)mRNA和蛋白、[磷酸化(p-)JAK2、p-STAT3]蛋白表达降低(P<0.05)。与GB组相比,inhibitor组细胞增殖率、乳酸含量、葡萄糖消耗水平、(Cyclin D1、PKM2)mRNA和蛋白表达进一步降低(P<0.05),activator组则显著逆转了上述指标的变化(P<0.05)。结论GB能够通过下调JAK2/STAT3信号通路抑制OE19细胞的增殖与糖酵解。Objective To investigate the effect and mechanism of ginkgolide B(GB)on the proliferation and glycolysis of esophageal cancer cells.Methods Human esophageal carcinoma OE19 was cultured in vitro.They were divided into control group(Con group)(no intervention),low/medium/high dose experimental group(L/M/H group)(12.5,25,50μmol·L^(-1)GB)and cisplatin(CP)group(4 mg·L^(-1)CP).The optimal concentration of GB was screened for subsequent experiments.Then they were divided into Con group,GB group(12.5μmol·L^(-1)GB),CP group,inhibitor group(12.5μmol·L^(-1)GB+10μmol·L^(-1)JAK2/STAT3 pathway inhibitor AG490),activator group(12.5μmol·L^(-1)GB+0.5μmol·L^(-1)JAK2/STAT3 pathway activator Colivelin)for 24 h.Cell counting kit-8(CCK-8),5-ethynyl-2′-deoxyuridine(EdU)method,lactate detection kit,glucose detection kit,real-time fluorescent quantitative PCR(RT-qPCR)and western blot(WB)were used to detect cell viability,proliferation rate,lactate content,glucose consumption level,and cyclin D1(Cyclin D1),M2 pyruvate kinase(PKM2)]mRNA and protein expression levels,Janus kinase 2(JAK2)/activator of transcription 3(STAT3)signal pathway related protein expression levels.Results Compared with the Con group,the cell viability of the L/M/H group and the CP group decreased(P<0.05),so in this study,the lower concentration of 12.5μmol·L^(-1)GB was selected as the GB group for subsequent experiments.Compared with the Con group,the cell proliferation rate,lactate content,glucose consumption level,mRNA and protein expression of Cyclin D1 and PKM2,and protein expression of phosphorylated JAK2 and p-STAT3 were decreased in GB and CP groups(P<0.05).Compared with the GB group,the cell proliferation rate,lactate content,glucose consumption level,Cyclin D1,PKM2 mRNA and protein expression in the inhibitor group were further decreased(P<0.05),and the activator group significantly reversed the changes in the above indicators(P<0.05).Conclusion GB can inhibit the proliferation and glycolysis of OE19 cells by down-regulating JAK2/STAT3 signal

关 键 词：食管癌 银杏内酯B Janus激酶2/信号转导和转录启动因子3 增殖 糖酵解 

分 类 号：R571[医药卫生—消化系统]