| | | 人硒蛋白P cDNA探针的制备及其在肝脏组织中的表达
| | 临床医学检验论文 作者：南克俊 隋晨光 韩王月 刘彦仿 胡沛臻 秦海霞 景钊 刘陕西
【关键词】 人硒蛋白P
Preparation of human selenoprotein P cDNA probe and expression in human hepatic tissue
【Abstract】 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 (χ2=15.84, P<0.005; χ2=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.
【Keywords】 human selenoprotein P; cDNA probes; PCR; in situ hybridization; carcinoma, hepatocellular
【摘要】 目的临床医学检验论文： 探讨SeP基因与肝脏疾病特别是肝癌发生发展的临床医学检验论文关系. 方法： 采用美国Georgia大学惠赠的质粒PBluescriptHuman Selenoprotein P，用PCR法制备人SeP cDNA探针，检测了13例正常肝脏、20例肝硬化、30例肝癌组织中SeP mRNA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SeP mRNA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SeP mRNA阳性率分别为84.6%，50.0%，20.0%. 肝癌细胞阳性表达率明显低于其他两组（χ2=15.84, P<0.005; χ2=4.96, P<0.05）. 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SeP cDNA探针制备成功，可用于人体组织SeP mRNA的原位检测. 肝癌细胞中存在SeP mRNA的表达缺失，SeP基因的缺失可能与肝癌的发生有关.
【关键词】 人硒蛋白P；cDNA探针；PCR；原位杂交；癌，肝细胞
0引言
硒是人体必需的微量元素之一，参与GSHPx的合成，具有抗氧化损伤作用［1］，流行病学调查表明缺硒可导致恶性肿瘤的发生［2］. 实验证实食物中添加硒可以减少前列腺癌、结肠癌、肺癌及肝癌的发生［3］. 硒的许多生物学功能是通过硒蛋白来实现的，迄今已经发现了18种具有生物功能的硒蛋白，硒蛋白P（Selenoprotein P，SeP）是其中一种［4］. 人体几乎所有的组织中都含有SeP，近年来大量研究证实其与多种恶性肿瘤有关，已经成为研究的热点. 但从基因水平对肝癌组织中的SeP进行原位检测，目前尚少见报道. 因此我们制备了特异性的SeP cDNA探针，对正常肝脏、肝硬化、肝癌组织进行核酸原位杂交，检测其表达水平，探讨SeP与肝脏疾病特别是肝癌发生发展的关系.
|
| | |
| | | | <<<<<全文未完>>>>> 全文字数约$$$zishu$$$字 | |
| | |
| |
|
教育论文中心