摘要
目的本研究通过对HBV相关肝细胞癌(HCC)患者血清N-聚糖检测及肝癌组织与癌旁组织中糖基转移酶基因表达水平比较分析,探索HCC患者血清N-聚糖变化的可能机制。方法收集解放军总医院2018年—2019年HBV相关HCC手术患者(34例)的肝癌和癌旁组织及正常肝组织标本,同时采集血清标本。从34例HCC患者中随机选择8例HCC患者血清标本作为HCC试验组,20例健康成年人血清标本作为对照组。应用DSA-FACE法分析HCC试验组与对照组血清N-聚糖图谱。采用荧光定量PCR法检测34例HBV相关HCC患者癌组织和癌旁组织中8种糖基转移酶基因(FUT3、FUT4、FUT6、FUT7、FUT8、Gn-TⅢ、Gn-TⅣa和Gn-TⅤ)mRNA表达水平,并应用蛋白印迹法检测相应蛋白表达水平。计量资料两组间比较采用独立样本t检验。结果与对照相比,8例HCC患者血清中N-聚糖峰9(peak9,NA3Fb)的丰度明显升高(t=-2.514,P<0.05)。糖基转移酶FUT8、Gn-TⅣa和Gn-TⅤ基因mRNA表达水平在癌组织和癌旁组织间有差异,其中癌组织中FUT8和Gn-TⅤ基因的mRNA与蛋白表达水平显著高于癌旁组织(mRNA:1.50±0.34 vs 0.65±0.11,t=-2.354,P=0.022;3.57±0.64 vs 1.33±0.16,t=-3.384,P=0.001)(蛋白:0.70±0.11 vs 0.083±0.017,t=9.555,P=0.001;1.33±0.19 vs 0.60±0.15,t=5.097,P=0.007);癌组织中Gn-TⅣa基因的mRNA表达水平显著高于癌旁组织(mRNA:2.90±0.47 vs 1.68±0.19,t=-2.403,P=0.019),蛋白表达水平与癌旁组织无显著差异(蛋白:0.52±0.24 vs 0.24±0.11,t=1.833,P=0.141)。癌组织中这些糖基转移酶表达改变与血清中N-聚糖丰度变化一致。结论HBV相关HCC患者血清中一些N-聚糖水平变化可能与肝癌组织中糖基转移酶基因GnT-V、GnT-IVa和FUT8表达上调密切相关。
Objective To investigate the potential mechanism of serum N-glycan alterations in patients with hepatitis B virus(HBV)-related hepatocellular carcinoma(HCC)by measuring serum N-glycan profile and comparing glycosyltransferase gene expression between HCC tissue and adjacent tissue.Methods The samples of HCC tissue,adjacent tissue,and normal liver tissue were collected from 34 patients with HBV-related HCC who were admitted to Chinese PLA General Hospital,and serum samples were also collected.Among these 34 patients,8 were randomly selected and their serum samples were established as HCC experimental group,and the serum samples of 20 healthy adults were established as control group.DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis was used to analyze serum N-glycan profile in the HCC experimental group and the control group.Quantitative real-time PCR was used to measure the mRNA expression of 8 glycosyltransferase genes(FUT3,FUT4,FUT6,FUT7,FUT8,Gn-TIII,Gn-TIVa,and Gn-TV)in the HCC tissue and adjacent tissue of 34 patients with HBV-related HCC,and Western blot was used to measure the expression of corresponding proteins.The independent samples t-test was used for comparison of continuous data between two groups.Results Compared with the control group,the HCC experimental group had a significant increase in the abundance of N-glycan peak9(NA3Fb)in serum(t=-2.514,P<0.05).There were significant differences in the mRNA expression of FUT8,Gn-TIVa,and Gn-TV between HCC tissue and adjacent tissue,and the mRNA and protein expression levels of FUT8 and Gn-TV in HCC tissue were significantly higher than those in adjacent tissue(FUT8 mRNA:1.50±0.34 vs 0.65±0.11,t=-2.354,P=0.022;Gn-TV mRNA:3.57±0.64 vs 1.33±0.16,t=-3.384,P=0.001;FUT8 protein:0.70±0.11 vs 0.083±0.017,t=9.555,P=0.001;Gn-TV protein:1.33±0.19 vs 0.60±0.15,t=5.097,P=0.007).The mRNA expression level of Gn-TIVa in HCC tissue was significantly higher than that in adjacent tissue(2.90±0.47 vs 1.68±0.19,t=-2.403,P=0.019),but there was no significant difference in the protein expression level of Gn-TIVa between HCC tissue and adjacent tissue(0.52±0.24 vs 0.24±0.11,t=1.833,P=0.141).The changes of glycosyltransferase gene expression in HCC tissue were consistent with the alteration of serum N-glycan profile.Conclusion Serum N-glycan alterations in patients with HBV-related HCC may be closely associated with the upregulated expression of the glycosyltransferase genes FUT8,Gn-TIVa,and Gn-TV in HCC tissue.
作者
曹曦
孙艳玲
陈翠英
肖义炜
向宽辉
刘学恩
庄辉
CAO Xi;SUN Yanling;CHEN Cuiying;XIAO Yiwei;XIANG Kuanhui;LIU Xueen;ZHUANG Hui(Department of Microbiology and Center of Infectious Diseases,School of Basic Medical Sciences,Peking University Health Science Center,Beijing 100191,China;Department of Hepatic Surgery,Liver Disease Center,The Fifth Medical Center of Chinese PLA General Hospital,Beijing 100853,China;Jiangsu Xiansida Biotechnology Co.,Ltd.,Nanjing 210000,China)
出处
《临床肝胆病杂志》
CAS
北大核心
2021年第6期1336-1341,共6页
Journal of Clinical Hepatology
基金
“十三五”国家重大科技专项(2018ZX10732401-003-015)
广西病毒性肝炎防治研究重点实验室开放课题基金项目(GXCDCKL201901)。
关键词
肝肿瘤
实验性
N-聚糖
糖基转移酶类
Liver Neoplasms,Experimental
N-glycan
Glycosyltransferases