PFKFB3在结直肠炎癌转化中的作用及作为治疗靶点的机制研究

Colitis is closely related to the occurrence of colon cancer. Control of colitis and inhibition of colorectal cancer cells proliferation are crucial strategies to the treatment of CAC. PFKFB3, which catalyzes the formation of fructose 2,6-diphosphate, is an important regulator of glycolysis. And the glycolysis is the main form of glucose metabolism for tumor cells and M1 macrophages. We found that the PFKFB3 inhibitor PFK15 could inhibit colitis and CAC in mice. Therefore, we hypothesize that inhibition of PFKFB3 suppresses colorectal tumorigenesis by inhibiting colitis and the malignant transformation of intestinal epithelial cells, restrains the development of colorectal cancer by inhibiting colorectal cancer cells proliferation and lactic acid production. To confirm this hypothesis, the following study will be conducted: 1) To establish mouse model with Pfkfb3 conditional knockout in macrophages and intestinal epithelial cells, to induce the colitis and CAC in this mouse model and observe the phenotypic differences between them. 2) To isolate macrophages, intestinal epithelial and colorectal cancer cells, and to explore the role and mechanism of PFKFB3-mediated glycolysis on key signaling pathways such as macrophage polarization, malignant transformation of intestinal epithelial cells and colorectal cancer cells proliferation. 3) To analyze the relationship between the expression-and-distribution of PFKFB3 and the prognosis of patients with colitis and CAC. This proposed study will investigate in depth the role of PFKFB3 in CAC and provide a new target for clinical diagnosis and treatment of this disease.

肠炎与肠癌密切相关，控制肠炎及抑制肠癌细胞增殖是治疗肠炎相关肠癌（CAC）的关键。催化形成果糖-2,6-二磷酸的PFKFB3是糖酵解过程的重要调控因子。肿瘤细胞、M1型巨噬细胞主要利用糖酵解方式进行葡萄糖代谢。我们发现PFKFB3的抑制剂PFK15可以抑制小鼠肠炎以及CAC。由此我们假设抑制PFKFB3可通过抑制肠炎抑制肠上皮细胞恶性转化从而抑制肠癌发生，并可通过抑制肠癌细胞增殖及乳酸产生从而抑制肠癌发展。为证实该假设，将开展以下研究：1）建立巨噬及肠上皮细胞Pfkfb3特异敲除小鼠，诱导小鼠发生肠炎与CAC，观察小鼠表型差异；2）分离巨噬、肠上皮及肠癌细胞，探索PFKFB3介导的糖酵解对巨噬细胞极化、肠上皮恶性转化、肠癌增殖等关键信号通路的作用及机制；3）分析PFKFB3表达分布与肠炎及CAC患者预后的关系。本研究将深入解析PFKFB3在CAC中的作用，为临床诊疗此疾病提供新的靶点。

肠炎与肠癌密切相关，控制肠炎及抑制肠癌细胞增殖是治疗肠炎相关肠癌（CAC）的关键 。我们前期研究发现PFKFB3的抑制剂PFK15可以抑制小鼠肠炎以及CAC。由此我们假设抑制PFKFB3可通过抑制肠炎抑制肠上皮细胞恶性转化从而抑制肠癌发生 ，并可通过抑制肠癌细胞增殖及乳酸产生从而抑制肠癌发展。为证实该假设，我们开展以下工作：1）建立巨噬及肠上皮细胞Pfkfb3特异敲除小鼠，诱导小鼠发生肠炎与CAC，观察小鼠表型差异；2）以SW480、HCT116,SW620三株结肠癌细胞株为体外模型证实抑制PFKFB3的表达明显降低这三株细胞株的存活率其机制可能与细胞自噬相关；3）：在AOM/DSS小鼠体外模型中也证实PFK-15抑制小鼠肠炎进展，并且抑制AOM/DSS肠癌模型小鼠的肿瘤进展；裸鼠移植瘤实验也证实了PFK-15对SW480细胞体内的增殖；4）：收集了55余例结直肠癌临床样本，分析PFKFB3表达分布与肠炎及CAC患者预后的关系。本研究将深入解析PFKFB3在CAC中的作用，为临床诊疗此疾病提供新的靶点和手段。