CXCR3通过调控Notch1信号影响乙肝病毒X蛋白对肝癌细胞周期和迁移的作用  

CXCR3 Influences the Effects of Hepatitis B Virus X Protein on the Cycle and Migration of Hepatocellular Carcinoma Cells by Regulating Notch1 Signaling

作　　者：胡泉东 周斌 李猛 杨玉娟 傅月美 HU Quandong;ZHOU Bin;LI Meng;YANG Yujuan;FU Yuemei(Shaoing Vocational and Technical College,Shaoring 312000,China;Department of Pathophysiology,School of Basic Medicine,Nanchang University,Nanchang 330006,China;Affiliated Hospital of Shaoxing College of Arts and Sciences,Shaoxing 312000,China)

机构地区：[1]绍兴职业技术学院,绍兴312000 [2]南昌大学基础医学院病理生理学教研室,南昌330006 [3]绍兴文理学院附属医院,绍兴312000

出　　处：《病毒学报》2022年第6期1382-1390,共9页Chinese Journal of Virology

基　　金：浙江省教育厅一般科研项目(项目号:Y202045015),题目:新时代高职护理专业突发公共卫生事件应急能力培养设计与研究。

摘　　要：乙肝病毒X蛋白(Hepatitis B virus X protein,HBx)是乙肝病毒诱导肝癌进展的关键因子,其能够影响肝癌细胞周期和促进其迁移。已知趋化因子受体3(chemokine CXC motif receptor 3,CXCR3)在乙型肝炎病毒肝癌中发挥促进作用,目前尚不清楚CXCR3在HBx诱导的肝癌细胞中的作用。因此,本研究探讨下调CXCR3对HBx诱导的肝癌细胞周期进展和迁移作用及机制。肝癌细胞Huh7分成对照组,pcDNA组(转染阴性对照载体),pcDNAHBx组(转染HBx过表达载体),pcDNA-HBx+si-NC组(共转染HBx过表达载体、siRNA对照的),pcDNA-HBx+si-CXCR3组(共转染HBx过表达载体、CXCR3 siRNA的),pcDNA-HBx+si-CXCR3+Jagged1组(共转染HBx过表达载体、CXCR3 siRNA,Notch1信号激活剂Jagged1处理),测定细胞中HBx、CXCR3、神经性钙黏附素(Neural cadherin,N-cadherin)、基质金属蛋白酶-9(Matrix metalloprotease 9,MMP-9)、细胞周期蛋白D1(cyclin D1)、增殖细胞核抗原(PCNA)、基质金属蛋白酶-2(Matrix metalloprotease 2,MMP-2)、上皮性钙黏附素(Epithelical cadherin,Ecadherin)、波形蛋白(Vimentin)、Notch神经同源蛋白1前体(Neurogenic locus notch homolog protein 1 precusor,Notch1)、Hes1蛋白表达,检测细胞增殖、周期和迁移、侵袭情况。结果表明,与Control、pcDNA组比较,pcDNAHBx组肝癌细胞中HBx、CXCR3、Notch1、Hes1、Cyclin D1、PCNA、MMP-9、MMP-2、vimentin、N-cadherin蛋白表达量升高,细胞增殖活性、迁移数目、侵袭数目升高,细胞G0/G1期比例、E-cadherin蛋白表达量降低(P<0.05);与pcDNA-HBx+si-NC组比较,pcDNA-HBx+si-CXCR3组肝癌细胞中CXCR3、Notch1、Hes1、Cyclin D1、PCNA、MMP-9、MMP-2、vimentin、N-cadherin蛋白表达量降低,细胞增殖活性、迁移数目、侵袭数目降低,细胞G0/G1期比例、E-cadherin蛋白表达量升高(P<0.05);与pcDNA-HBx+si-CXCR3组比较,pcDNA-HBx+si-CXCR3+Jagged1组肝癌细胞Notch1、Hes1、Cyclin D1、PCNA、MMP-9、MMP-2、vimentin、N-cadherin蛋白表达量升高,细胞增殖活性、迁Hepatitis B virus X protein(HBx)is a key factor for the hepatitis B virus to induce hepatocellular carcinoma(HCC)progression.HBx can affect the cycle and promote the migration of HCC cells.Chemokine CXC motif receptor 3(CXCR3)plays a part in HBV infection progressing to HCC,but the mechanism of CXCR3 in HBx-induced HCC is unclear.Therefore,we investigated the effect and mechanism of action of HBxinduced cycle progression and migration of HCCs.A HCC line(Huh7)was employed.We divided Huh7 cells into five groups:control;pcDNA(transfected with a negative control vector),pcDNA-HBx(transfected with an HBx overexpression vector);pcDNA-HBx+si-NC group(co-transfected with an HBx overexpression vector and siRNA control),pcDNA-HBx+si-CXCR3(co-transfected with an HBx overexpression vector and CXCR3 siRNA),pcDNA-HBx+si-CXCR3+Jagged1(co-transfected with an HBx overexpression vector,CXCR3 siRNA,and Notch1 signal activator Jagged1).We measured protein expression of HBx,CXCR3,neural cadherin(N-cadherin),matrix metalloprotease-9(MMP-9),cyclin D1,proliferating cell nuclear antigen(PCNA),MMP-2,epithelial cadherin(E-cadherin),vimentin,Notch neural homolog protein 1 precursor(Notch1),and Hes1,and assessed the proliferation,cycle,migration,and invasion of HCC cells.Compared with control and pcDNA groups,expression of HBx,CXCR3,Notch1,Hes1,cyclin D1,PCNA,MMP-9,MMP-2,vimentin,and N-cadherin in the pcDNA-HBx group of HCC cells increased,cell proliferation,the number of migrating cells,and number of invading cells increased,and the ratio of cells in the G0/G1 phase,and protein expression of E-cadherin decreased(P<0.05).Compared with the pcDNA-HBx+si-NC group,protein expression of CXCR3,Notch1,Hes1,cyclin D1,PCNA,MMP-9,MMP-2,vimentin,and N-cadherin in HCC cells decreased,cell proliferation,the number of migrating cells,and number of invading cells decreased,and the ratio of the number of cells in the G0/G1 phase,and protein expression of E-cadherin increased(P<0.05)in the pcDNA-HBx+si-CXCR3 group.Compared with the pcDNA-HBx+si-CXCR3group,pr

关 键 词：乙肝病毒X蛋白 肝癌 CXCR3 NOTCH1 周期 迁移 

分 类 号：R373.21[医药卫生—病原生物学]