mTOR信号通路在调控卵巢癌CD4+Treg糖代谢中的作用
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R737.31

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国家自然科学基金(81772779);深圳市宝安区科技创新计划基础研究项目(2020JD462)


Role of mTOR signaling pathway in regulating CD4 + Treg glucose metabolism in ovarian cancer
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    摘要:

    目的:初步探讨mTOR 信号通路在卵巢癌患者CD4+ Treg(CD4+ CD25+ CD127low调节性T 细胞)糖代谢过程中的作用。方法:流式细胞术检测卵巢癌患者(ovarian cancer,OC)、卵巢良性肿瘤患者(benign ovarian tumor,BOT)以及健康对照者 (healthy controls,HC)外周血CD4+ Treg中mTOR+ 细胞的百分比;建立CD4+ Treg与SKOV3共培养体系,检测共培养前后mTOR+ 的CD4+ Treg百分比,使用雷帕霉素抑制mTOR信号后,采用比色法检测CD4+ Treg细胞糖摄取和糖酵解水平,采用实时荧光定量PCR(real-time fluorescence quantitative PCR,RT-PCR)和Western blot 检测CD4+ Treg 细胞糖代谢相关基因和蛋白的表达水平。结果:卵巢癌患者外周血中 mTOR+ 的CD4+ Treg百分比显著高于健康对照组[(77.4±8.12)% vs(. 64.19±9.7)%,P<0.01),与 SKOV3共培养后CD4+ Treg中mTOR+ 细胞百分比明显升高(P<0.05),mTOR信号通路相关基因和蛋白表达水平也升高;抑制 SKOV3生长环境中CD4+ Treg的mTOR信号后其糖代谢相关基因和蛋白表达水平显著下降,糖摄取(193.49±13.28 vs. 174.05± 14.58,P<0.01)和糖酵解水平(21.97±0.87 vs. 4.85±1.54,P<0.001)也明显下降。结论:卵巢癌患者CD4+ Treg中mTOR及其下游信号通路明显活化,mTOR信号通路可通过调节CD4+ Treg的糖摄取和糖酵解水平,并调控糖代谢相关基因和蛋白的表达水平,影响CD4+ Treg的糖代谢进程。

    Abstract:

    Objective:To explore the role of mTOR signaling pathway in CD4 + regulatory T cell(Treg)glucose metabolism in ovarian cancer patients. Methods:Flow cytometry was used to detect the percentage of mTOR+ CD4 + Treg in peripheral blood with ovarian cancer(OC)patients,benign ovarian tumor(BOT)patients and healthy controls(HC). We established a coculture system of human CD4+ Treg with ovarian cancer cell SKOV3,and detected the percentage of mTOR+ CD4+ Treg before and after co-culture. After mTOR signal was inhibited by rapamycin,the glucose uptake and glycolysis levels of CD4+ Treg were detected by colorimetry,and the expression levels of genes and proteins related to glucose metabolism in CD4 + Treg were detected by real-time quantitative PCR and Western blot. Results:The percentage of mTOR+ CD4+ Treg in peripheral blood of ovarian cancer patients was significantly higher than that of healthy controls[(77.4±8.12)% vs(. 64.19±9.7)%,P < 0.01]. After co-culture with SKOV3,the percentage of mTOR+ CD4 + Treg were elevated(P < 0.05),and the levels of mTOR signaling pathway related genes and proteins also increased. After inhibition of mTOR signal of CD4 + Treg in SKOV3 growth environment,the glucose uptake(193.49 ± 13.28 vs. 174.05 ± 14.58,P < 0.01)and glycolysis level(21.97±0.87 vs. 4.85±1.54,P < 0.001)were decreased significantly. Glucose metabolism - related genes and proteins were also significantly reduced. Conclusion:mTOR and its downstream signaling pathway are significantly activated in peripheral blood CD4 + Treg of ovarian cancer patients,and mTOR signaling pathway can affect glucose metabolism of CD4 + Treg by regulating glucose metabolist-related genes and protein levels.

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付鑫,吴茗,陈献,刘书娜,张磊,李荣,徐娟,陶子琦,王婷,王芳. mTOR信号通路在调控卵巢癌CD4+Treg糖代谢中的作用[J].南京医科大学学报(自然科学版),2023,(5):604-610

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  • 收稿日期:2022-12-13
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  • 在线发布日期: 2023-05-18
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