mTOR信号通路在调控卵巢癌CD4+ Tregs糖代谢中的作用
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1.南京医科大学第一附属医院检验学部,江苏南京 210029;2.南京医科大学第一附属医院检验学部;3.广东省深圳市宝安区妇幼保健院检验科

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国家自然科学基金项目(面上项目81772779);广东省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+ Tregs glucose metabolism in ovarian cancer
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1.Department of Laboratory Medicine, First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu Province 210029;2.Department of Laboratory Medicine, First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu Province

Fund Project:

The National Natural Science Foundation of China (no. 81772779), Baoan District Research project of Shenzhen,(2020JD462)

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    摘要:

    摘要:目的:初步探讨mTOR信号通路在卵巢癌患者CD4+ Tregs(CD4+ CD25+ CD127low Regulatory T cells)糖代谢过程中的作用。方法:流式细胞术(Flow Cytometry,FCM)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+ Tregs中mTOR+ 细胞的百分比;建立CD4+ Tregs与SKOV3共培养体系,检测共培养前后mTOR+ 的CD4+ Tregs百分比,使用雷帕霉素抑制mTOR信号后,采用比色法检测CD4+ Tregs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WB(Western blot)检测CD4+ Tregs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+ Tregs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+ Tregs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+ Tregs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+ Tregs中mTOR及其下游信号通路明显活化,mTOR信号通路可通过调控糖代谢相关基因和蛋白水平来影响CD4+ Tregs的糖代谢进程。

    Abstract:

    Abstract:objective: To explore the role of mTOR signaling pathway in CD4+ Tregs glucose metabolism in ovarian cancer patients. Methods: Flow cytometry was used to detect the percentage of mTOR+ CD4+ Tregs cells 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+ Tregs cells with ovarian cancer cell SKOV3, and detect the percentage of mTOR+ CD4+ Tregs before and after co-culture. After mTOR signal was inhibited by rapamycin, the glucose uptake and glycolysis levels of CD4+ Tregs cells were detected by colorimetry, and the expression levels of genes and proteins related to glucose metabolism in CD4+ Tregs cells were detected by real-time quantitative PCR and Western blot. Results: The percentage of mTOR+CD4+ Tregs 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+ Tregs cells 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+ Tregs 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; Conclusions: mTOR and its downstream signaling pathway are significantly activated in peripheral blood CD4+ Tregs of ovarian cancer patients, and mTOR signaling pathway can affect glucose metabolism of CD4+ Tregs by regulating glucose metabolist-related genes and protein levels.

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  • 收稿日期:2022-06-16
  • 最后修改日期:2022-09-20
  • 录用日期:2023-05-11
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