文章摘要
张晓洁,吴 梦,陈 献,娄鉴芳,黄珮珺,黄 蕾,孙瑞红,王 芳.mTORC1-HIF1α通路基因在人CD8+调节T细胞中的表达及意义[J].南京医科大学学报,2016,(12):1418~1421,1480
mTORC1-HIF1α通路基因在人CD8+调节T细胞中的表达及意义
Expressions and significances of mTORC1-HIF1α pathway genes in human CD8+ regulatory T cell
投稿时间:2016-06-13  
DOI:10.7655/NYDXBNS20161202
中文关键词: 卵巢癌  CD8+T细胞  调节性T细胞  mTORC1-HIF1α通路
英文关键词: ovarian cancer cell  CD8+T cell  regulatory T cell  mTORC1-HIF1α pathway
基金项目:
作者单位
张晓洁 南京医科大学第一附属医院检验学部,江苏 南京 210029
江苏省妇幼保健院检验科,江苏 南京 210036 
吴 梦 南京医科大学第一附属医院检验学部,江苏 南京 210029 
陈 献 南京医科大学第一附属医院检验学部,江苏 南京 210029 
娄鉴芳 南京医科大学第一附属医院检验学部,江苏 南京 210029 
黄珮珺 南京医科大学第一附属医院检验学部,江苏 南京 210029 
黄 蕾 南京医科大学第一附属医院检验学部,江苏 南京 210029 
孙瑞红 南京医科大学第一附属医院检验学部,江苏 南京 210029 
王 芳 南京医科大学第一附属医院检验学部,江苏 南京 210029 
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中文摘要:
      目的:探讨mTORC1-HIF1α通路基因在人CD8+调节性T细胞中的表达及意义?方法:建立人卵巢癌细胞系SKOV3与健康人CD8+ T细胞体外共培养体系,设置CD8+T细胞单独培养组为对照组?共培养5 d后,收集各组CD8+T细胞,荧光定量PCR检测2组CD8+T细胞中mTORC1-HIF1α通路基因(mTORC1?HIF1α?Glut1?PKM2?GPI?TPI?Eno1及LDHα)mRNA表达水平;Western blot 检测2组CD8+T细胞中mTORC1-HIF1α通路基因(mTORC1?HIF1α及PKM2)蛋白表达水平;收集14例卵巢癌,12例卵巢良性肿瘤及12例健康体检者外周血,磁珠阳选法分离CD8+T细胞,荧光定量PCR检测卵巢癌患者CD8+T细胞中mTORC1-HIF1α通路基因mRNA表达水平,并与卵巢良性肿瘤及健康体检者做比较研究?结果:与单独培养组相比,共培养组CD8+ T细胞mTORC1?HIF1α?PKM2?GPI及TPI mRNA表达水平显著降低(P < 0.05);Western blot 结果显示,共培养组CD8+ T细胞mTORC1?HIF1α及PKM2蛋白表达水平也显著低于对照组(P < 0.05);卵巢癌组CD8+ T细胞中mTORC1?HIF1α?Glut1?PKM2?GPI及TPI表达量显著低于健康对照组(P < 0.05);卵巢癌组mTORC1?PKM2?GPI及TPI表达量明显低于卵巢良性肿瘤组(P < 0.05);卵巢良性组mTORC1-HIF1α通路各基因表达水平与健康对照组间无显著性差异?结论:卵巢癌微环境诱导的CD8+调节性T细胞低表达mTORC1-HIF1α通路基因,mTORC1-HIF1α通路在卵巢癌微环境中CD8+调节性T细胞代谢及分化过程中具有重要意义?
英文摘要:
      Objective:To investigate the expressions and significances of mTORC1-HIF1α pathway genes in human CD8+ regulatory T cell. Methods:Coculture systerm of CD8+ T cells and SKOV3 were conducted in vitro,CD8+ T cells cultured alone group acted as control group. At day 5,CD8+ T cells were collected,and used to examine the expression of eight glycolysis genes by quantitative real-time reverse transcriptase(qRT)-PCR(mTORC1,HIF1α,Glut1,PKM2,GPI,TPI,Eno1,and LDHα). Western blot was used to detect the expression of mTORC1,HIF1α and PKM2. We collected peripheral blood from 14 ovarian cancer(OC) patients,12 benign diseases,and 12 healthy females. CD8+ T cells were then separated using a CD8-positive isolation kit. The expressions of mTORC1,HIF1α,Glut1,PKM2,GPI,TPI,Eno1,and LDHα in CD8+ T cells from OC patients was detected by qRT-PCR and compared with that in patients with ovary benign tumor(BOT)and healthy volunteers. Results:Compared with CD8+ T cells cultured without SKOV3 cells,glycolysis gene expression showed varying degrees of decline in CD8+ T cells cultured with SKOV3 cells. The expression of mTORC1,HIF1α,PKM2,GPI,and TPI was significantly decreased in co-cultured cells compared with the control group. Results showed that the expression of mTORC1,HIF1α,and PKM2 decreased significantly(P < 0.05)in CD8+ T cells cultured with SKOV3 cells compared with control group. HIF1α and Glut1 mRNA had lower expression levels in CD8+ T cells from OC patients than those from healthy controls(both P < 0.05). mTORC1,PKM2,GPI,and TPI were also expressed at lower levels in OC patients than those in either BOT patients or healthy controls(both P < 0.05). The expression of mTORC1-HIF1α pathway genes has no difference between benign diseases group and the healthy controls. Conclusion:mTORC1-HIF1α signalling integrates the control of CD8+ T cells metabolism and differentiation in ovarian cancer microenvironment,which play a significant role in the immunotherapy of OC
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