p75NTR在细胞核和细胞浆不同部位表达与食管癌细胞分化程度的关系及其作用研究

项目名称： p75NTR在细胞核和细胞浆不同部位表达与食管癌细胞分化程度的关系及其作用研究

项目编号： No.31460310

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 生物科学

项目作者： 慕晓玲

作者单位： 石河子大学

项目金额： 50万元

中文摘要： 肿瘤干细胞是晚期癌症死亡率高的主要原因，认识肿瘤干细胞有助于进一步了解肿瘤细胞的生物学特点，发展直接靶向肿瘤干细胞的治疗技术。但是有关食管癌干细胞生物学知之甚少。基于前期研究结果，我们提出食管癌细胞中p75NTR核阳性表达的细胞可能就是食管肿瘤干细胞或分化程度比较低的干细胞的假设。本项目将通过采用单细胞克隆和细胞球培养技术分别从食管癌细胞株和原代培养的细胞中分离p75NTR核阳性表达细胞克隆和p75NTR浆阳性表达细胞克隆，采用荧光免疫细胞技术、Western-blot技术和PCR技术，分析两种细胞克隆的食管上皮干细胞标签蛋白表达情况，采用MTT、平板克隆、Transwell和体内种植致瘤检测其增殖和自我更新能力、侵袭性和致瘤能力，证明p75NTR核阳性表达细胞克隆是食管癌干细胞或分化程度比较低的干细胞，并在此基础上探讨p75NTR对维持食管癌干细胞的生存及自我更新的作用及机制。

中文关键词： 食管癌干细胞；p75NTR；核表达；干细胞分化；自我更新

英文摘要： Cancer stem cells are the leading cause of high cancer mortality for advanced cancer. Therefore, there is an increasing need to understand the biology of cancer stem cells so that new therapeutic developments could tackle this cell population. However, comparatively little is known about the biology of esophageal cancer stem cells (ECSC) compared to other solid tumor stem cells because the special marker of ECSC has not been found. We suppose a hypothesis that the cell expressing p75NTR in nucleus is likely ECSC or the more primitive ECSC in the hierarchy of esophageal cancer stem cell, on account of the results we have done. So in this grant,we will separate the cell clones of p75NTR+ expression in nucleus and in cytoplasm by single cell clone and cell spheres formation from the human esophageal cancer cell line and the primary culture esophageal cancer cells of patient, respectively, to detect its expression of esophageal epithelial stem cells marker with immunofluorescent cell technology ,Western-blot and PcR,respectively; to analysis their proliferation, self-renewal, invasion and tumorigenesis by MTT, Flat clone, Transwell and xenografted into immunodeficient mice, respectively, to prove the hypothesis we supposed.

英文关键词： esophageal cancer stem cell;p75NTR;expression in nuclear;stem cell differentiation;self-renewal