Abstract

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.

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