人硒蛋白PcDNA探针的制备及其在肝脏组织中的表达

Preparation of human selenoprotein P cDNA probe and expression in human hepatic tissue

目的 :探讨SeP基因与肝脏疾病特别是肝癌发生发展的关系 .方法 :采用美国Georgia大学惠赠的质粒PBluescript HumanSelenoproteinP,用PCR法制备人SePcDNA探针 ,检测了 13例正常肝脏、2 0例肝硬化、30例肝癌组织中SePmR NA的表达水平 .结果 :PCR产物经电泳鉴定与预期相符 .3种肝脏组织的组织间质均有SePmRNA表达 ,主要位于脉管区的血管内皮 ;正常肝细胞、肝硬化肝细胞及肝癌细胞中SePmRNA阳性率分别为 84 .6 % ,5 0 .0 % ,2 0 .0 % .肝癌细胞阳性表达率明显低于其他两组 (χ2 =15 .84 ,P <0 .0 0 5 ;χ2 =4 .96 ,P <0 .0 5 ) .正常肝细胞和肝硬化肝细胞的胞质及胞核内均出现蓝色的阳性颗粒 ,在胞核主要分布在核仁与核周 ;正常肝细胞的核阳性强于肝硬化肝细胞 .HCC细胞阳性表达颗粒位于胞质 ,胞核几无表达 .结论 :人SePcDNA探针制备成功 ,可用于人体组织SePmRNA的原位检测 .肝癌细胞中存在SePmRNA的表达缺失 。

AIM: To investigate the expression of selenoprotein P mRNA in hepatic diseases, including cirrhosis and hepatocellular carcinoma (HCC). METHODS: Selenoprotein P cDNA fragment was amplified by PCR the expression of Selenoprotein P mRNA was detected in 13 cases of normal liver, 20 cases of cirrhosis and 30 cases of hepatocellular carcinoma (HCC) by in situ hybridization. RESULTS: The PCR products evaluated by electrophoresis were as expected. The expression of selenoprotein P mRNA was found in the tissue stroma of liver, especially in the endothelial cells of vasculature. The positive expression rate of selenoprotein P mRNA was 84.6% (11/13), 50.0% (10/20), 20.0% (6/30) respectively in normal liver, cirrhosis and HCC. The rate of expression in hepatic carcinoma was significantly lower than that in others (χ 2=15.84, P < 0.005 ; χ 2=4.96, P <0.05). The positive blue granules were seen distributed in the nucleus and cytoplasm in normal liver and cirrhosis liver. The expression in nucleus of normal liver was significantly higher than that of cirrhosis liver. In HCC, the positive particles were located in cytoplasm, but nucleolus showed almost no staining. CONCLUSION : The expression of human selenoprotein P gene is significant different in the three kinds of liver tissues, suggesting that selenoprotein P may play a role in the occurrence and development of HCC.