外源突变型p53和潜伏膜蛋白1基因在转基因鼠不同器官组织中的表达  

Expression of exogenous mutant p53 and latent membrane protein 1 genes in the tissues of the different organs of transgenic mice

作　　者：何迎春[1] 田道法[1] 田甜[1] 卢芳国[1] 刘宇勤[1] 

机构地区：[1]湖南中医学院基础部,湖南省长沙市410007

出　　处：《中国临床康复》2005年第42期99-101,共3页Chinese Journal of Clinical Rehabilitation

基　　金：中国博士后科学基金第35批资助项目(No.2004035644);湖南省杰出青年基金项目(No.03JJY3040);湖南省教育厅基金项目(No.03C287)~~

摘　　要：目的:观察转基因小鼠鼻腔、鼻咽、肺、食道、胃、大肠、肝、肾、脾中外源突变型p53m t和潜伏膜蛋白1基因的表达情况。方法:实验于2005-03/04在湖南中医学院中西医结合系基础研究室完成。选取4月龄转p53m t和潜伏膜蛋白1基因F0代小鼠1只,以正常同龄未转基因C57BL/6J小鼠4只作为对照。采用免疫组化分别检测转基因鼠和对照鼠鼻腔、鼻咽、肺、食道、胃、大肠、肝、肾、脾中p53m t和潜伏膜蛋白1基因的表达强度,用图像分析系统测定阳性反应物的平均灰度值。系统设定白色最大灰度为256,黑色最小灰度为0。灰度值越小,代表阳性反映程度越高。结果:实验纳入转基因鼠1只,对照鼠4只,均进入结果分析。①蛋白质阳性表达信号的定位分布:转基因鼠p53m t和潜伏膜蛋白1基因的免疫组化阳性信号均为棕黄色的颗粒或斑块,表达量高,p53m t蛋白绝大部分分布于细胞膜和胞浆内,少数位于细胞核中。对照鼠p53mt蛋白在组织中呈弱表达,潜伏膜蛋白1的蛋白阳性信号主要位于上皮细胞的细胞膜和胞浆中。②转基因鼠和对照鼠不同组织部位中p53m t基因表达的平均灰度值:转基因鼠食道、肝、脾中平均灰度值较高(131.75±17.25,135.15±13.14,139.42±11.55),在鼻腔黏膜、鼻咽组织、大肠中平均灰度值较低(96.61±12.43,109.26±9.90,109.45±6.72)。除了胃、脾外,各组织部位与对照鼠比较均有显著差异(P<0.05)。③转基因鼠和对照鼠不同组织部位中潜伏膜蛋白1基因表达的平均灰度值:转基因鼠鼻咽部平均灰度值最低(106.75±19.57),其次为鼻腔、食道、肺、胃,而在大肠、肝、脾中表达水平较高(152.29±12.17,134.98±17.79,158.35±10.75)。除鼻腔、大肠、肝、脾外,各组织部位中潜伏膜蛋白1基因的表达水平与对照鼠比较均有显著差异(P<0.05)。④转基因鼠不同组织部位中p53mt基因与潜伏膜蛋白1基因表达水平的相关性�AIM： To investigate the expression of exogenous mutant p53（p53mt） and latent membrane protein 1 （LMP1）genes transferred into C57BL/6J mice in the tissues of different organs collected from these transgenic animals, such as the tissues of nasal cavity, nasopharynx, lung, esophagus, stomach, large intestine, liver, kidney and spleen,METHODS： The experiment was carried out from March to April 2005 in the Basic Laboratory of the Department of Integrated Traditional Chinese and Western Medicine, Hunan University of Traditional Chinese Medicine. The experiment was carried out in one Fo transgenic mouse carrying p53mt and LMP1 genes aged 4 months, and four non-transgenic C57BL/6J mice of 4 months as controls. The tissue samples were collected respectively from the different organs of the experimental mice to detect the expression of p53mt and LMPI with immunohistochemical method and to analyze the expression difference of those two genes among different organs, such as the tissues of nasal cavity, nasopharynx, lung, esophagus, stomach, large intestine, liver, kidney and spleen. Then, the average gray values were measured by image analysis system. Value of 256 was considered as the highest gray value, 0 as the lowest. The lower the gray value, the higher the positive reaction. RESULTS： All the experimental mice were enrolled in the result analysis. ①Positional distribution of positive signal of protein expression： Immunohistochemical positive signals of p53mt and LMPI in the transgenic mouse were brown granules or plaques. Most of p53mt proteins were found in cell membrane and cytoplasm, and few in nuclei. Expression of p53mt was weak in the non-transgenic mice, and positive LMP1 expression was mainly found in cell membrane and cytoplasm of epithelial cells. ②Mean gray value of p53mt： There were the lower expression levels of p53mt in esophagus （ 131.75±17.25 ）, liver （ 135.15±13.14） and spleen （139.42±11.55）, and the higher expression levels in nasal mucosa（96.61±12.43 ）, nasop

关 键 词：转基因 蛋白质P53 病毒包膜蛋白质类 基因表达 

分 类 号：R349.64[医药卫生—基础医学]