共生环境下内皮细胞糖酵解靶向阻断对膀胱癌血管生成的影响及机制

Currently, the block effect of angiogenesis is still not satisfying. Some researches even reported that this therapy may increase the risk of tumor recurrence and metastasis. Our previous proteomic studies discovered the expressions of fructose 6-diphosphate kinase 2 (PFK2) and monocarboxylate transporter 1 (MCT1) were increased in endothelial cells of bladder cancer which indicated that the glycolysis of endothelial cells was enhanced. Whether there are connections among aerobic glycolysis of cancer, glycolysis function of endothelial cells and angiogenesis remain unknown. Based on our findings, we speculate that there were some metabolic coupling between cancer cells and endothelial cells. The possible mechanism is that cancer cells undergo aerobic glycolysis and release lactate, lactate enters endothelial cells through MCT1. The increased lactate concentration directly leads to angiogenesis, meanwhile, the glycolysis of endothelial cells is enhanced and may offer energy to angiogenesis. The targeted therapy should be stable and effective if the deprivation of energy of endothelial cells is available, and should obtain additional advantages for this method can escape from the complicated regulation mechanism of angiogenesis. In order to verify our speculation, we designed a proper microfluidic chip platform to investigate metabolic coupling between cancer cells and endothelial cells. Based on microfluidic chip platform, the driving mechanism and subsequent metabolic changes in endothelial cells under a co-culture environment were discussed. Furthermore, to offer more evidences to metabolic-coupling and angiogenesis, our study included glycolysis, mitochondrial function, etc. What's more, combined target small interfering technique was performed to block energy generation as well as MCT1 by in vitro and in vivo experiments. Above all things, present research item will help to explain the mechanism of bladder cancer and lay the foundation for the novel targeted antitumor therapy.

目前靶向血管生成疗效并不理想，甚至会增加肿瘤复发转移的风险。前期研究中我们发现膀胱癌血管内皮细胞6磷酸果糖激酶2(PFK2)、单羧酸转运体1(MCT1)表达上调，提示内皮细胞糖酵解功能增强。膀胱癌细胞的有氧酵解与内皮细胞糖酵解及血管生成之间是否存在关联？为此我们提出假说"肿瘤细胞糖酵解产生大量乳酸，乳酸通过MCT1进入内皮细胞，诱发血管生成，并上调糖酵解功能为血管生成提供能量。如能阻断内皮细胞的能量代谢，可以绕开血管生成的复杂调控机制，使靶向治疗稳定，有效"。为验证这一假说，我们拟建立适合本实验条件的高通量微流控芯片实验室；在此基础上探讨共生环境下血管生成的驱动机制以及后续代谢改变，从糖酵解、线粒体功能等多方面为血管生成特点及代谢模式提供佐证，并利用联合靶向性小干扰RNA技术阻断乳酸转运通路及糖酵解限速酶，探讨其体内外抗血管生成作用，为阐明膀胱癌的发病机制及靶向治疗夯实理论基础。

肿瘤血管生成与肿瘤进展密切相关，目前临床靶向血管生成疗效并不理想，甚至会增加肿瘤复发转移的风险。我们前期研究发现膀胱癌血管内皮细胞6磷酸果糖激酶2(PFK2)、单羧酸转运体1(MCT1)表达上调，提示内皮细胞糖酵解功能增强，推测膀胱癌细胞的有氧酵解与内皮细胞糖酵解及血管生成之间密切关联。为此我们建立适合本实验条件的高通量微流控芯片实验室；在此基础上探讨共生环境下血管生成的驱动机制以及后续代谢改变，从糖酵解、线粒体功能等多方面为血管生成特点及代谢模式提供佐证，并利用联合靶向性小干扰RNA技术阻断乳酸转运通路及糖酵解限速酶，探讨其体内外抗血管生成作用，从而证实肿瘤细胞糖酵解产生的大量乳酸，可以通过MCT1进入内皮细胞，诱发血管生成，并上调糖酵解功能为血管生成提供能量，同时我们阻断内皮细胞的能量代谢，可以绕开血管生成的复杂调控机制，使靶向治疗稳定，有效，从而为研究膀胱癌的发病机制及临床靶向治疗提供新思路。