卵巢癌细胞生长环境中CD4^(+)Treg细胞和CD4^(+)Teff细胞的糖代谢特征初探  

The glucose metabolism characteristics of CD4^(+)Treg cells and CD4^(+)Teff cells in growth environment of ovarian cancer cells

作　　者：吴茗 徐睿 刘书娜 王芳[1] WU Ming;XU Rui;LIU Shuna;WANG Fang(Department of Laboratory Medicine,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China)

机构地区：[1]南京医科大学第一附属医院检验学部,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2021年第7期999-1005,共7页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金(81772779);江苏省“六大人才高峰”资助项目(2015-WSN-034);江苏省“六个一工程”拔尖人才项目(LGY2017068);江苏省重点人才基金(ZDRCA2016003);江苏省实验诊断学重点实验室(ZDXKB2016005)。

摘　　要：目的:探究CD4^(+)CD25+CD127low调节性T细胞(regulatory T cell,Treg)和CD4^(+)CD25-CD127+效应性T细胞(effector T cell,Teff)在卵巢癌细胞生长环境中的糖代谢特征。方法:流式细胞术(fluorescence-activated cell sorting,FACS)分选CD4^(+)Treg细胞和CD4^(+)Teff细胞,建立两种细胞与SKOV3细胞的共培养体系,采用实时荧光定量PCR(real-time fluorescence quantitative PCR,RT-PCR)和Western blot检测CD4^(+)Treg细胞、CD4^(+)Teff细胞糖代谢相关基因和蛋白的表达水平,比色法检测细胞糖摄取和糖酵解水平。结果:SKOV3共培养组CD4^(+)Treg细胞糖代谢相关基因和蛋白的表达水平高于CD4^(+)Teff细胞和CD4^(+)Treg细胞单独培养组(P<0.05);SKOV3共培养组CD4^(+)Treg细胞糖摄取和糖酵解水平高于CD4^(+)Teff细胞[(59.78±2.57)pmol vs.(44.93±1.86)pmol,P<0.05;(15.31±2.05)mmol/L vs.(7.98±0.88)mmol/L,P<0.05]。结论:卵巢癌细胞生长环境中的人外周血CD4^(+)Treg细胞糖代谢水平高于CD4^(+)Teff细胞,卵巢癌细胞可以促进人外周血CD4^(+)Treg细胞糖代谢相关蛋白和基因表达水平上调。Objective:To investigate the glucose metabolism characteristics of CD4^(+)CD25+CD127low regulatory T cells(Treg)and CD4^(+)CD25-CD127+ effector T cells(Teff)in growth environment of ovarian cancer cells. Methods:Fluorescence-activated cell sorting(FACS)separated CD4^(+)Treg cells and CD4^(+)Teff cells,then established a coculture system of human T cells with ovarian cancer cell SKOV3. The expression levels of glucose metabolism related genes and proteins were determined by real-time fluorescence quantitative PCR(RT-PCR)and Western blot. The levels of glucose uptake and glycolysis were detected by colorimetry. Results:Compared with the CD4^(+)Treg cells in single culture without SKOV3 and CD4^(+)Teff cells,the expression levels of glucose metabolism related genes and proteins in CD4^(+)Treg cells cocultured with SKOV3 were elevated(P<0.05)and the levels of glucose uptake and glycolysis were increased[(59.78±2.57)pmol vs.(44.93±1.86)pmol,P<0.05;(15.31 ± 2.05)mmol/L vs.(7.98 ±0.88)mmol/L,P<0.05]. Conclusion:The level of glucose metabolism in CD4^(+)Treg cells is higher than CD4^(+)Teff cells in ovarian cancer cell growth environment,ovarian cancer cells could promote the expression levels of glucose metabolism related genes and proteins in CD4^(+)Treg cells.

关 键 词：卵巢癌 调节性T细胞 效应性T细胞 糖代谢 

分 类 号：R737.31[医药卫生—肿瘤]