mTORC1-HIF1α通路基因在人CD8+调节T细胞中的表达及意义
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Expressions and significances of mTORC1-HIF1α pathway genes in human CD8+ regulatory T cell
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    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-

    Abstract:

    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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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(自然科学版),2016,(12):1418-1421,1480

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  • 收稿日期:2016-06-13
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  • 在线发布日期: 2016-12-28
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