中国生化药物杂志
中國生化藥物雜誌
중국생화약물잡지
Chinese Journal of Biochemical Pharmaceutics
2015年
10期
5-10
,共6页
丙型肝炎病毒%唾液酸转移酶%半乳糖基转移酶%HCV感染%糖苷标志物
丙型肝炎病毒%唾液痠轉移酶%半乳糖基轉移酶%HCV感染%糖苷標誌物
병형간염병독%타액산전이매%반유당기전이매%HCV감염%당감표지물
hepatitis C virus%sialyltransferase%galactosyltransferase%HCV infection%biomarker
目的 探讨丙型肝炎病毒(hepatitis C virus,HCV)感染人肝癌细胞系Huh7.5.1细胞后与肿瘤相关的唾液酸转移酶家族和半乳糖基转移酶家族的35个糖基转移酶的表达变化.方法 利用HCV感染的Huh7.5.1肝细胞模型,采用Western blot检测HCV感染后HCV非结构蛋白NS3的表达,定量即时聚合酶链锁反应(quantitative real-time polymerase chain reaction,qRT-PCR)检测HCV感染后HCV-RNA的表达及HCV感染对Huh7.5.1细胞中35个糖基转移酶mRNA表达水平的影响.凝集素染色实验测定HCV感染对Huh7.5.1细胞中糖苷表达水平的影响.结果 在HCV感染后的Huh7.5.1 细胞中,发现4 个唾液酸转移酶(ST3Gal Ⅲ、ST6GalNAC Ⅲ、ST8Sia Ⅲ和ST8Sia Ⅳ)和5个半乳糖基转移酶(B3GALT 1、B3GALT 2、B3GALT 3、B3GALT 4和B4GALNT 2)的mRNA水平明显升高11~45倍,其中变化最大的是B3GALT 1,上升45倍,其次是ST8Sia Ⅲ和ST8Sia Ⅳ,分别上升37倍和39倍.进一步采用凝集素染色实验验证,发现结合末端唾液酸糖苷Neu5Acα6Gal的接骨木凝集素(elderberry lectin,EBL)在HCV感染后升高1.97倍,结合末端半乳糖苷Galβ1,3GalNAc的尾穗苋凝集素(amaranthus caudatus agglutinin,ACA)和结合Galβ1,4GalNAc的鸡冠刺桐凝集素(erythrina cristagalli agglutinin,ECA)分别升高3倍和4.3倍.结论 HCV感染导致人肝细胞中特定唾液酸转移酶和半乳糖基转移酶及其相应糖苷表达水平呈现明显上升.这些结果对研究HCV感染的治疗靶点和诊断标志物具有一定参考价值.
目的 探討丙型肝炎病毒(hepatitis C virus,HCV)感染人肝癌細胞繫Huh7.5.1細胞後與腫瘤相關的唾液痠轉移酶傢族和半乳糖基轉移酶傢族的35箇糖基轉移酶的錶達變化.方法 利用HCV感染的Huh7.5.1肝細胞模型,採用Western blot檢測HCV感染後HCV非結構蛋白NS3的錶達,定量即時聚閤酶鏈鎖反應(quantitative real-time polymerase chain reaction,qRT-PCR)檢測HCV感染後HCV-RNA的錶達及HCV感染對Huh7.5.1細胞中35箇糖基轉移酶mRNA錶達水平的影響.凝集素染色實驗測定HCV感染對Huh7.5.1細胞中糖苷錶達水平的影響.結果 在HCV感染後的Huh7.5.1 細胞中,髮現4 箇唾液痠轉移酶(ST3Gal Ⅲ、ST6GalNAC Ⅲ、ST8Sia Ⅲ和ST8Sia Ⅳ)和5箇半乳糖基轉移酶(B3GALT 1、B3GALT 2、B3GALT 3、B3GALT 4和B4GALNT 2)的mRNA水平明顯升高11~45倍,其中變化最大的是B3GALT 1,上升45倍,其次是ST8Sia Ⅲ和ST8Sia Ⅳ,分彆上升37倍和39倍.進一步採用凝集素染色實驗驗證,髮現結閤末耑唾液痠糖苷Neu5Acα6Gal的接骨木凝集素(elderberry lectin,EBL)在HCV感染後升高1.97倍,結閤末耑半乳糖苷Galβ1,3GalNAc的尾穗莧凝集素(amaranthus caudatus agglutinin,ACA)和結閤Galβ1,4GalNAc的鷄冠刺桐凝集素(erythrina cristagalli agglutinin,ECA)分彆升高3倍和4.3倍.結論 HCV感染導緻人肝細胞中特定唾液痠轉移酶和半乳糖基轉移酶及其相應糖苷錶達水平呈現明顯上升.這些結果對研究HCV感染的治療靶點和診斷標誌物具有一定參攷價值.
목적 탐토병형간염병독(hepatitis C virus,HCV)감염인간암세포계Huh7.5.1세포후여종류상관적타액산전이매가족화반유당기전이매가족적35개당기전이매적표체변화.방법 이용HCV감염적Huh7.5.1간세포모형,채용Western blot검측HCV감염후HCV비결구단백NS3적표체,정량즉시취합매련쇄반응(quantitative real-time polymerase chain reaction,qRT-PCR)검측HCV감염후HCV-RNA적표체급HCV감염대Huh7.5.1세포중35개당기전이매mRNA표체수평적영향.응집소염색실험측정HCV감염대Huh7.5.1세포중당감표체수평적영향.결과 재HCV감염후적Huh7.5.1 세포중,발현4 개타액산전이매(ST3Gal Ⅲ、ST6GalNAC Ⅲ、ST8Sia Ⅲ화ST8Sia Ⅳ)화5개반유당기전이매(B3GALT 1、B3GALT 2、B3GALT 3、B3GALT 4화B4GALNT 2)적mRNA수평명현승고11~45배,기중변화최대적시B3GALT 1,상승45배,기차시ST8Sia Ⅲ화ST8Sia Ⅳ,분별상승37배화39배.진일보채용응집소염색실험험증,발현결합말단타액산당감Neu5Acα6Gal적접골목응집소(elderberry lectin,EBL)재HCV감염후승고1.97배,결합말단반유당감Galβ1,3GalNAc적미수현응집소(amaranthus caudatus agglutinin,ACA)화결합Galβ1,4GalNAc적계관자동응집소(erythrina cristagalli agglutinin,ECA)분별승고3배화4.3배.결론 HCV감염도치인간세포중특정타액산전이매화반유당기전이매급기상응당감표체수평정현명현상승.저사결과대연구HCV감염적치료파점화진단표지물구유일정삼고개치.
Objective To investigate the altered glycotransferases and glycan profile in HCV-infected cells, the expression levels of 35 kinds of sialyltransferases and galactosyltransferases were evaluated in hepatitis C virus (HCV) infected Huh7.5.1 cell cultured model.Methods Western blot was used to measure the expressions of HCV-nonstructural protein NS3 and and quantitative real-time PCR (qRT-PCR) was performed to determine HCV-RNA levels and mRNA levels of 35 kinds of sialyltransferases and galactosyltransferases.Lectin microarray was used to compare the glycan profiles of sialyltransferases-and galactosyltransferases-associated glycan-profile in Huh7.5.1 cells and HCV infected Huh7.5.1 cells.Results Among the 35 kinds of sialyltransferases and galactosyltransferases, the mRNA levels of four sialyltransferases (including ST3Gal Ⅲ, ST6GalNAC Ⅲ, ST8Sia Ⅲ and ST8SiaⅣ) and five galactosyltransferases (including B3GALT1, B3GALT 2, B3GALT3, B3GALT4 and B4GALNT2) were found to be 11-45 fold higher in HCV-infected cells compared with naive Huh7.5.1 cells.The up-regulated level of B3GALT 1 was the most evident (45-fold change), followed by ST8Sia Ⅳ and ST8Sia Ⅲ, with 39-and 37-fold respectively.Consistently, lectin microarray showed that glycans recognized by EBL (binding terminal sialic acid, Neu5Acα6Gal, 1.97-fold change), ECA letin (binding terminal galactose, Galβ1,4GalNAc,4.3-fold change), and ACA(binding terminal galactose, Galβ1-3GalNAc, 3-fold change) were elevated in glycoprotein products of sialyltransferase and galactosyltransferase respectively.Conclusion HCV infection causes the increased expression levels of mutiple sialyltransferases and galactosyltransferases in Huh7.5.1 cell line and hence the upregulation of glycan profiles of the glycoproteins.These results provide potential therapeutic targets and diagnostic markers for HCV infection and insights on the molecular mechanism of HCV infection.
