新狼毒素A通过NAD+补救信号通路调节能量代谢逆转黑色素瘤恶性表型的机制研究

Our research group have examined the anti-proliferation effects of a series of chemical compounds isolated from Stellera chamaejasme L. in melanoma cells. We found that neochamaejasmin A (NCA) could inhibit NAMPT mRNA expression and effect cellular NAD+ level, which accompanied by down-regulation of key enzyme of glycolysis (GLUT1, HK2, PFK-1) mRNA level and raise mitochondrial respiration, leading to cancer cell proliferative inhibition and malignant phenotypes reverse. In this study, two tumor cell lines (mouse melanoma B16F0 and human melanoma A375) and human normal melanocytes will be used as the research object. We will define how NAD+ salvage pathway regulates cancer cell energy metabolism in NCA-treated melanoma cells and clarify the role of NCA in the regulation of tumor cell energy metabolism. We will further find out that the regulation of NCA on cancer cell energy metabolism to inhibit the proliferation of tumor cells and reverse the malignant phenotype. Meanwhile, non-targeted metabonomics analysis will used to explore the differential metabolites and differential pathways of the malignant phenotype of melanoma induced by NCA. These results could guide the chemical structural modification of NCA, and find more active new compounds, which supplying a basis for the study of NCA and providing a useful attempt to screen the natural products for targeting to tumor energy metabolism.

课题组对狼毒的系列化学成分抗黑色素瘤增殖的作用进行了初步研究，发现新狼毒素A能够下调黑色素瘤细胞内NAD+水平和NAMPT mRNA及蛋白表达，黑色素瘤细胞内糖酵解关键酶（GLUT1, HK2, PFK-1）水平明显下降，线粒体呼吸作用增强，使得黑色素瘤细胞增殖抑制和恶性表型发生逆转。本研究拟采用小鼠黑色素瘤B16F0、人黑色素瘤A375，以及人正常黑色素细胞为研究对象，旨在弄清新狼毒素A如何影响肿瘤细胞NAD+补救通路，明确其对肿瘤细胞能量代谢的调控作用；进一步明确新狼毒素A干扰肿瘤细胞能量代谢从而抑制肿瘤细胞增殖、逆转恶性表型的作用机制。同时利用非靶向代谢组学分析方法，研究新狼毒素A使黑色素瘤恶性表型逆转后出现的差异代谢物以及差异通路。研究结果可指导新狼毒素A化学结构改造，找到活性更强的新化合物，从而为新狼毒素A类似物的研究提供基础，为筛选靶向肿瘤能量代谢的天然产物研究提供有益尝试。

本课题对新狼毒素A及其有关类似物抗黑色素瘤增殖的作用进行了较为系统研究，发现新狼毒素A对小鼠黑色素瘤B16F0、人黑色素瘤A375有明显的抑制作用，但对人正常黑色素细胞的抑制作用不明显，作用机制研究表明新狼毒素A通过一系列途径影响黑色素瘤细胞NAD+补救通路，并进一步对肿瘤细胞能量代谢起到调控作用；并明确了新狼毒素A干扰肿瘤细胞能量代谢，及其抑制肿瘤细胞增殖、逆转恶性表型的作用机制。同时通过基因测序分析方法，找到了新狼毒素A使黑色素瘤恶性表型逆转后出现的差异代谢物以及差异通路。