淋巴细胞HLA-A mRNA方法学建立及在乙型肝炎病毒感染患者中的应用  

The development of methodology for HLA-A mRNA in lymphocytes and its application in patients with HBV infection

作　　者：王传新[1] 郑桂喜[1] 邹雄[1] 李晓丽[1] 王金凤[1] 刘敏[1] 于修平[2] 

机构地区：[1]山东大学齐鲁医院检验科,250012 [2]山东大学医学院病原微生物教研室

出　　处：《中华检验医学杂志》2006年第8期708-710,共3页Chinese Journal of Laboratory Medicine

基　　金：山东省科技厅资助项目(2002BB1 CJA8)

摘　　要：目的建立外周血淋巴细胞HLA-A mRNA的检测方法,探讨其在慢性乙型肝炎、肝炎后肝硬化、肝癌患者中的应用价值。方法改良法提取人外周血淋巴细胞RNA,应用逆转录-聚合酶链反应(RT-PCR)一步法扩增HLA-A mRNA,琼脂糖凝胶电泳结果采用Bandscan软件分析相对灰度值,半定量HLA-A的基因表达。结果半定量HLA-A所得平均相对灰度值在慢性乙型肝炎组、肝硬化组、肝癌组、正常对照组分别为0．54±0．21、0．34±0．27、0．22±0．16、0．95±0．19。慢性乙型肝炎组、肝硬化组、肝癌组均明显低于正常对照组(P<0．05),慢性乙型肝炎组与肝癌组,肝硬化组与肝癌组相比差异有统计学意义(P<0．05),慢性乙型肝炎组与肝硬化组有下降趋势。结论成功建立检测HLA-A基因的RT-PCR一步法,从基因转录水平上证实慢性乙型肝炎、肝炎后肝硬化、肝癌患者外周血淋巴细胞HLA-A下调,且与相应患者细胞免疫功能减弱密切相关。Objective To establish a method for the detection of HLA-A mRNA in peripheral blood lymphocytes and investigates its applicable value in patients with hepatitis B, hepatocirrhosis, hepatocelluar carcinoma. Methods We used the advanced method to extract the RNA of peripheral blood lymphocytes. Then we amplificated HLA-A mRNA by one-step RT-PCR. The amplified fragments were analyzed by Bandscan software after agarose gel electrophoresis to semi-quantify the expression of HLA-A. Results The average relative gray value acquired by semi-quantify the expression of HLA-A in hepatitis B patients, hepatocirrhosis patients, hepatocelluar carcinoma patients, and the normal control were 0. 54 ± 0. 21,0. 34 ± 0. 27,0. 22 ± 0. 16, and 0. 95 ± 0. 19 respectively. The expression of HLA-A in the groups of hepatitis B patients,hepatocirrhosis patients,hepatocelluar carcinoma patients were all significantly lower than that in the controls （ P 〈 0. 05 ） . Only the expression of HLA-A in hepatitis B and hepatocirrhosis patients were significantly lower than that in hepatocelluar carcinoma patients. Only the expression of HLA-A between hepatitis B and hepatocirrhosis patients demonstrate a downtrend. Conclusions The method of detecting HLA-A gene by RT-PCR was successfully established. We confirmed HLA-A significantly declined in peripheral blood lymphocytes of patients with hepatitis B, hepatocirrhosis, hepatocelluar carcinoma on the level of gene translation. The expression of HLA-A was closely correlated with the state of cell immunity.

关 键 词：HLA-A抗原 逆转录聚合酶链反应 肝炎 乙型 慢性 肝硬化 肝肿瘤 

分 类 号：R512.62[医药卫生—内科学]