雌激素受体β调控CD4+ T细胞介导的肿瘤微环境促进膀胱癌进展及分子机制

Proliferation and invasion are main cause of death of bladder cancer (BCa). It is reported that both occurrence and biological behavior of BCa are related to genders, and estrogen receptor β could promote invasion of BCa. Tumor microenvironment (TME) could also impact BCa’s biological behaviors, our previous study demonstrated that CD4+ T cell could promote invasion of BCa. In this project, we are going to study ERβ’s role in CD4+ T cell conducted TME and its potential mechanisms. Our preliminary test suggested that, (1) CD4+ T cell could promote proliferation and invasion of BCa, as well as expression of ERβ and c-MET; (2) blocking ERβ could partially reverse CD4+ T cell promoted BCa invasion and c-MET expression; (3) blocking ERβ could also reduce the number of CD4+ T cells recruited to BCa in vitro. Considering preliminary results above, we propose a hypothesis that CD4+ T cell promotes c-MET via ERβ, and promoted ERβ could further recuit CD4+ T cells to BCa, thus promotes BCa progression as a positive feedback loop. This project aims to study ERβ’s role in CD4+ T cell conducted TME, as well as the mechanism how it modulates proliferation and invasion related key genes, via gene transfection, gene interfering, ChIP, luciferase reporter assay and etc..

膀胱癌的增殖和侵袭是其致死主因。文献报导膀胱癌的生物学行为与性别相关，雌激素受体β可促进膀胱癌的增殖和侵袭。肿瘤微环境对于膀胱癌的生物学行为亦有作用，我们前期发现CD4+ T细胞可促进膀胱癌的侵袭。因此我们拟研究雌激素受体β(ERβ)在CD4+ T细胞介导的肿瘤微环境中的作用及机制。预实验结果表明，CD4+ T细胞可提高膀胱癌的增殖、侵袭及ERβ、c-MET的表达，阻断ERβ可逆转CD4+ T细胞引起的侵袭力增加及c-MET的表达升高，还可致CD4+ T细胞募集至膀胱癌的数目减少。因此我们提出假说，在此微环境中CD4+ T细胞可通过提升ERβ来增加c-MET表达，而ERβ的升高可进一步增加膀胱癌对CD4+ T细胞的募集，正反馈式促进膀胱癌的进展。本课题拟通过基因转染、干扰、染色质免疫沉淀、荧光素酶报告基因实验等技术，研究ERβ在该肿瘤微环境中的作用及其对肿瘤增殖、侵袭的关键基因的调控机制。

膀胱癌的增殖和侵袭是其致死主因。文献报导膀胱癌的生物学行为与性别相关，雌激素受体β可促进膀胱癌的增殖和侵袭。肿瘤微环境对于膀胱癌的生物学行为亦有作用，我们前期发现CD4+ T细胞可促进膀胱癌的侵袭。因此我们拟研究雌激素受体β(ERβ)在CD4+ T细胞介导的肿瘤微环境中的作用及机制。实验结果表明，CD4+ T细胞可提高膀胱癌的增殖、侵袭及ERβ、c-MET的表达，阻断ERβ可逆转CD4+ T细胞引起的侵袭力增加及c-MET 的表达升高，还可致CD4+ T细胞募集至膀胱癌的数目减少。在此微环境中，CD4+ T细胞可通过提升ERβ来增加c-MET表达，而ERβ的升高可进一步增加膀胱癌对CD4+ T细胞的募集，正反馈式促进膀胱癌的进展。本课题通过基因转染、干扰、染色质 免疫沉淀、荧光素酶报告基因实验等技术，发现浸润的 CD4+ T 细胞可以通过ERβ/c-MET和ERβ/IL-1/c-MET信号通路来增强膀胱癌细胞的增殖和转移。.在研究过程中我们还发现，NRBP1蛋白在膀胱肿瘤的增殖、凋亡等生物学行为中发挥重要作用。我们通过免疫组织化学和临床资料随访，发现NRBP1的高表达和膀胱癌的预后呈负相关；通过si-RNA沉默NRBP1基因，采用CCK-8和流式细胞术证实其可在体外实验中抑制肿瘤的生长，并促进其凋亡；随后我们在裸鼠成瘤实验中证实了si-NRBP1可抑制肿瘤生长。.本项目为通过雌激素信号通路控制膀胱肿瘤进展的潜在应用提供了一定的理论基础。其主要意义在于，为膀胱肿瘤的治疗提供了新思路，同时内分泌治疗可利用现有药物调控性激素水平实现肿瘤控制，省去大量新药研发费用，具有潜在的卫生经济学效益。