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）m RNA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m RNA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 m RNA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（q RT）-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 q RTPCR 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 m RNA 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.