过表达CD47通过阻断Fas/FasL途径抑制SW480人结肠癌细胞凋亡  

Overexpression of CD47 inhibits apoptosis of SW480 human colon cancer cells by blocking Fas/FasL pathway

作　　者：王芳[1] 吴晓婷 路志国 唐翠 曹相玫 WANG Fang;WU Xiaoting;LU Zhiguo;TANG Cui;CAO Xiangmei(Department of Gastroenterology,Ningxia Medical University General Hospital,Yinchuan 750004;School of Clinical Medicine,School of Basic Medicine,Ningxia Medical University,Yinchuan 750004,China;Department of Pathology,School of Basic Medicine,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学总医院消化内科,宁夏银川750004 [2]宁夏医科大学临床医学院,宁夏银川750004 [3]宁夏医科大学基础医学院病理学系,宁夏银川750004

出　　处：《细胞与分子免疫学杂志》2021年第10期904-909,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81860442,81560501)。

摘　　要：目的研究过表达CD47对SW480人结肠癌细胞凋亡的影响及可能机制。方法癌症基因组图谱(TCGA)数据库分析CD47在34种肿瘤中的表达,以及与结肠癌组织中免疫细胞浸润和细胞凋亡、黏附、血管生成等生物学过程的相关性。利用过表达CD47的慢病毒感染SW480细胞,实验分为空白对照组、空载体病毒感染组和CD47过表达组,通过荧光显微镜和Western blot法检测各组感染效率,流式细胞术检测细胞凋亡率,Western blot法检测凋亡相关蛋白自杀相关因子配体(FasL)、胱天蛋白酶8(caspase-8)、caspase-3的表达。结果TCGA数据库分析结果显示CD47在包括结肠癌在内的多种肿瘤中高表达,其mRNA水平与结肠癌微环境中CD4^(+)T细胞、CD8^(+)T细胞、B细胞、巨噬细胞、中性粒细胞的浸润相关,并参与细胞凋亡、黏附、血管生成等生物学过程,且蛋白互作网络提示CD47与死亡受体途径的相关蛋白存在相互作用。与空白对照组、空载体病毒感染组相比,过表达CD47的SW480细胞凋亡率显著下降,FasL、caspase-8、caspase-3蛋白表达均明显降低。结论CD47与结肠癌组织内免疫细胞浸润相关,且在结肠癌中高表达,过表达CD47可能通过阻断Fas/FasL途径抑制人结肠癌SW480细胞凋亡。Objectives To study the effect of overexpression of CD47 on apoptosis of human colon cancer SW480 cells and its underlying mechanism.Methods The expression of CD47 in thirty-four tumors was analyzed,along with its correlation to immune cell infiltration,apoptosis,adhesion and angiogenesis in colon cancer by referring to the TCGA database.Meanwhile we packaged an lentivirus vector expressing CD47 and successfully transfected SW480 cells with this vector to stably express CD47.These cells were divided into blank control group,empty body virus infection group and CD47 overexpression group,and then used for the following assays.The infection efficiency was detected by fluorescence microscope and Western blot analysis;the apoptosis rate was detected by flow cytometry;the expression of FasL,caspase-8 and caspase-3 was detected by Western blot analysis.Results The TCGA database analysis showed that CD47 was overexpressed in a variety of tumors,including colon cancer.The mRNA level of CD47 was related to the infiltration of CD4^(+)T cells,CD8^(+)T cells,B cells,macrophages and neutrophils in the colon cancer microenvironment.The expression of CD47 involved in biological processes such as apoptosis,adhesion and angiogenesis.Furthermore,the protein-protein interaction network suggested that CD47 interacted with proteins related to death receptor pathway.The apoptosis rate of SW480 cells and the expression of FasL,Caspase-8 and caspase-3 protein in overexpressing CD47 group was significantly higher than the blank control group and empty vector virus infection group.Conclusion The CD47 is highly expressed and is associated with immune cell infiltration in colon cancer.Overexpression of CD47 may inhibit apoptosis by blocking Fas/FasL pathway in human colon cancer SW480 cells.

关 键 词：CD47 结肠癌 SW480细胞 细胞凋亡 自杀相关因子配体(FasL) 

分 类 号：R735.35[医药卫生—肿瘤] Q279[医药卫生—临床医学]