肿瘤微环境缺氧诱导UHRF1上调介导Th17/Treg失衡促进髓母细胞瘤进展的研究

Recently, ubiquitin-like with PHD and RING finger domains 1(UHRF1) was revealed as an oncogene. We previously demonstrated that UHRF1 promoted medulloblastoma proliferation and progression. We recently revealed the expression of UHRF1,TGF-β1 and IL-10 was up-regulated under hypoxia circumstances. RNAi of UHRF1 inhibited TGF-β1 and IL-10 expression. The supernatants of medulloblastoma cells under hypoxia contained significantly more TGF-β1 and IL-10 protein, and the supernatants could lead to Treg cells expension. Combined with previous literature that reported Th17/Treg imbalance promoted tumor progression, we plan to：①use immunohistochemical staining,etc.method to detect and analyze the correlation of HIF-1α，UHRF1 and Th17/Treg cell frequency in medulloblastoma tissues and the relationship of previous biomarkers to clinical characteristics and prognosis of medulloblastoma. ②use microRNA microarray, Chip technology to study the mechanism of hypoxia inducing UHRF1 up-regulation, use MeDIP-seq technology to study the mechanism of UHRF1 up-regulating TGF-β1and IL-10. ③study the influence of TGF-β1and IL-10 to initiating CD4+T cells and Th17/Treg imbalance at cellular level, verify that UHRF1 inducing Th17/Treg imbalance and promoting tumor progression using animal model.

近年来，泛素样含PHD和环指域1(UHRF1) 被研究证实为癌基因。我们前期证实：UHRF1促进髓母细胞瘤增殖和进展。我们近期发现缺氧环境下髓母细胞中UHRF1、TGF-β1、IL-10表达上调，干扰UHRF1抑制TGF-β1、IL-10表达，缺氧诱导的髓母细胞培养上清中TGF-β1和IL-10明显升高，且上清可刺激Treg细胞扩增。结合文献报道Th17/Treg失衡促进肿瘤进展，本项目拟：①免疫组化等分析髓母组织HIF-1α，UHRF1与Th17/Treg的关联性，以及它们与肿瘤预后的关系。②microRNA芯片、Chip等技术研究缺氧诱导UHRF1上调的机制；MeDIP-seq技术研究UHRF1上调TGF-β1和IL-10的机制。③细胞水平研究TGF-β1和IL-10对CD4+T细胞分化的影响，介导Th17/Treg失衡；动物模型验证UHRF1介导Th17/Treg失衡，促进肿瘤进展。

近年来，泛素样含PHD和环指域1(UHRF1) 被研究证实为癌基因。我们发现UHRF1 在髓母细胞瘤中呈高表达，其表达水平影响髓母细胞瘤细胞系的增殖和进展。但UHRF1 促进髓母细胞瘤发生发展的具体机制不详。我们发现缺氧环境下髓母细胞中UHRF1、TGF-β1、IL-10表达上调，干扰UHRF1抑制TGF-β1、IL-10表达，缺氧诱导的髓母细胞培养上清中TGF-β1和IL-10明显升高，且上清可刺激Treg细胞扩增。本研究表明了髓母组织HIF-1α，UHRF1与Th17/Treg之间呈正相关。HIF-1α、UHRF1的高表达对预后不利；microRNA芯片、Chip等技术证实了缺氧诱导UHRF1上调；MeDIP-seq技术研究证实UHRF1上调TGF-β1和IL-10；细胞水平研究TGF-β1和IL-10对CD4+T细胞分化的影响，介导Th17/Treg失衡；动物模型验证UHRF1介导Th17/Treg失衡，促进肿瘤进展。本项目采用了免疫学、生物信息学、实验动物学以及分子生物学多层次阐释肿瘤微环境缺氧诱导UHRF1 上调介导Th17/Treg 失衡促进髓母细胞瘤进展及机制。从临床角度而言，相关问题的探讨有助于对UHRF1 促进肿瘤免疫逃逸机制的深入认识，有望为肿瘤的基因治疗、免疫治疗提供新思路。