肿瘤微环境中成纤维细胞自噬对膀胱癌糖代谢及恶性潜能的影响

Recent studies have focused on the relationship between tumor microenvironment, autophagy and carcinogenesis. However, little is known about the role of autophagy in a tumor microenvironment in tumor metabolism, and its underlying mechanisms. .Our previous researches have observed that fibroblasts could transformed into cancer-associated fibroblasts (CAFs) in a vivo-like microenvironment, accompanied with up-regulated expression of Monocarboxylate anion transporters (MCT) and enhanced autophagy, as well as elevated lactate levels in the tumor microenvironment. MCT inhibition of CAFs could suppress bladder cancer T24 cell proliferation and invasion, and induce cell apoptosis. In addition, we observed an interaction between p62 and MCT. Based on these results, we hypothesize that autophagy in CAFs might play a role in the development of bladder cancer in the tumor microenvironment, where glucose metabolism change might be involved. Consequently, we aim to understand whether CAFs can affect MCT expression and lactate levels, and subsequent glucose metabolism and biological behaviors of bladder tumor cells in the tumor microenvironment through autophagy, by means of 3-dimensional microfluidic co-culture device, lentivirus infection, pharmacological interventions, nude mouse tumorigenicity assay, et al. Finally, we would like to reveal the detailed role of autophagy in carcinogenesis from a new perspective of how autophagy takes effect on tumor in the microenvironment, and provide evidence of potential therapeutic targets.

肿瘤微环境和自噬是肿瘤领域的研究热点，而自噬对微环境中细胞代谢的影响研究较少。我们前期观察到共培养环境中成纤维细胞（hFF）可转化为肿瘤相关成纤维细胞，其单羧酸转运体MCT的表达上调，微环境中乳酸水平升高；抑制hFF的MCT表达可抑制膀胱癌细胞增殖和侵袭，诱导其凋亡，同时hFF自噬水平升高，该信号通路重要分子p62可与MCT相互作用，推断hFF自噬可能参与了膀胱癌的发生发展，尤其对细胞糖代谢产生影响。基于此假说，我们拟通过构建3D细胞共培养体系，采用分子克隆、药物干预、裸鼠成瘤实验等手段，通过改变微环境中hFF的自噬水平，从分子、细胞、体外和体内探讨hFF是否通过自噬-p62-MCT通路调节微环境乳酸水平，并进一步影响膀胱癌细胞糖代谢及生物学表型。本研究从肿瘤微环境中hFF自噬对微环境及膀胱癌糖代谢及生物学表型的作用这个新的视角出发，为揭示膀胱癌发病机制、探索临床可行治疗靶点提供新依据。

1、项目背景：肿瘤微环境和自噬是肿瘤领域的研究热点，而自噬对微环境中细胞代谢的影响研究较少。基于前期实验结果，我们推断成纤维细胞自噬可能参与了膀胱癌的发生发展，尤其对细胞糖代谢产生影响。基于此假说，我们探讨成纤维细胞是否通过自噬-MCT通路调节微环境乳酸水平，并进一步影响膀胱癌细胞糖代谢及生物学表型。.2、主要研究内容：（1）检测了MCT1在膀胱癌组织中的表达，并分析其表达与临床病理因素之间的相关性；并利用TCGA数据库进行验证；（2）在人膀胱癌T24细胞系中研究了MCT1对膀胱癌增殖、迁移、侵袭的影响；（3）体外研究了MCT1对膀胱癌EMT和糖代谢的影响；并初步揭示了HIF-1α在该过程中的作用；（4）体外诱导/抑制了成纤维细胞的自噬；（5）研究共培养环境中成纤维细胞自噬是否影响了T24细胞的生物学表型；（6）研究成纤维细胞自噬是否影响了T24细胞的糖代谢。.3、重要结果：（1）MCT1表达与膀胱癌预后有关；（2）MCT1可在体外和体内抑制膀胱癌增殖、迁移和侵袭；（3）MCT1可影响膀胱癌细胞糖代谢，并且在T24细胞中， MCT1表达与HIF-1α具有相关性；（4）成功诱导了成纤维细胞的自噬；（5）共培养环境中，成纤维细胞自噬影响了T24细胞增殖和侵袭能力；（6）共培养环境中，成纤维细胞自噬影响了T24细胞的糖代谢。.4、科学意义：本项目通过共培养体系，从分子、细胞、体外和体内探讨成纤维细胞是否通过自噬-MCT通路调节微环境乳酸水平，并进一步影响膀胱癌细胞糖代谢及生物学表型，为揭示膀胱癌发病机制、探索临床可行治疗靶点提供新依据。