胃癌细胞系中CD44^+胃癌干细胞的表达和意义  

Expression and Significance of CD44^+Gastric Cancer Stem Cells in Gastric Cancer Cell Lines

作　　者：任芳 陈毅德 冯丽华 郑志高 范鑫 林雨标 冯水土 REN Fang;CHEN Yi-de;FENG Li-hua;ZHENG Zhi-gao;FAN Xin;LIN Yu-biao;FENG Shui-tu(Department of Oncology and Hematology,Xiamen Haicang Hospital,Xiamen 361026,Fujian,China)

机构地区：[1]厦门市海沧医院肿瘤血液内科,福建厦门361026

出　　处：《医学信息》2020年第20期45-49,53,共6页Journal of Medical Information

基　　金：厦门市科技计划(医疗卫生项目)(编号:3502Z20184049、3502Z20199106);厦门市海沧区科技计划项目(编号:350205Z20174005)。

摘　　要：目的探讨胃癌细胞系中CD44^+胃癌干细胞的基因表达及其意义。方法采用无血清培养基获得漂浮球,在胃腺癌细胞系MKN-45中用FACS检测肿瘤干细胞(CSC)表面标志物CD44的表达,采用FACS分离CD44^+亚群,通过体外培养鉴定两种不同细胞群CD44^+和CD44-中的致瘤、自我更新、分化特性,采用实时RT-PCR评估CD44^+和CD44-中干细胞特异性基因的表达。结果MKN-45和NCI-N87在无血清培养基中形成胃癌微球体,而KATOⅢ和AGS不能形成胃癌微球体,MKN-45细胞系经FACS分选CD44^+和CD44-中细胞,80%MKN-45细胞高表达CD44;CD44^+细胞在无血清培养基中显示出更高的微球体形成、分化特性;参与Wnt2、Bmi1、Oct3/4、Notch1、Sox2、Nanog和其他基因的“干细胞”基因的表达水平在CD44^+亚群中高于CD44-亚群。结论人胃癌细胞中的CD44^+亚群可能含有胃癌干细胞样细胞,可为胃癌的诊断、治疗提供参考。Objective To investigate the gene expression and significance of CD44^+ gastric cancer stem cells in gastric cancer cell lines.MethodsFloating spheres were obtained with serum-free medium. FACS was used to detect the expression of tumor stem cell (CSC) surface marker CD44 ingastric adenocarcinoma cell line MKN-45, CD44^+ subgroups were separated by FACS, and two different cell populations were identified by in vitroculture The tumorigenicity, self-renewal, and differentiation characteristics of CD44^+ and CD44-. Real-time RT-PCR was used to evaluate theexpression of stem cell-specific genes in CD44^+ and CD44-.Results MKN-45 and NCI-N87 formed gastric cancer microspheres in serum-freemedium, while KATO and AGS could not form gastric cancer microspheres. The MKN-45 cell line was sorted by FACS for CD44^+ and CD44- cells,80% MKN-45 cells highly express CD44;CD44^+ cells show higher microsphere formation and differentiation characteristics in serum-free medium;the expression level of "stem cell" genes involved in Wnt2, Bmi1, Oct3/4, Notch1, Sox2, Nanog and other genes CD44^+ subgroup was higher thanCD44- subgroup.Conclusion The CD44^+ subset of human gastric cancer cells might contain gastric cancer stem cell-like cells, which could providereferences for the diagnosis and treatment of gastric cancer.

关 键 词：胃癌 CD44 胃癌干细胞 基因表达 

分 类 号：R735.2[医药卫生—肿瘤]