异甘草素调控糖代谢诱导黑色素瘤细胞凋亡机制研究

项目名称： 异甘草素调控糖代谢诱导黑色素瘤细胞凋亡机制研究

项目编号： No.31471338

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 生物科学

项目作者： 郑秋生

作者单位： 滨州医学院

项目金额： 90万元

中文摘要： 课题组前期研究表明，异甘草素能诱导黑色素瘤细胞凋亡，并伴随低氧诱导因子（HIF-1α）下调，葡萄糖摄取量降低，乳酸含量降低，ATP和活性氧（ROS）水平升高，线粒体电势去极化等现象，推测异甘草素可能靶向HIF-1，下调丙酮酸脱氢酶激酶1（PDK1） 表达，降低有氧糖酵解水平，上调氧化磷酸化代谢途径，ROS生成增多，活化线粒体凋亡途径或者产生内质网胁迫引起细胞凋亡。为验证上述假设，本研究拟采用小鼠黑色素瘤B16F0、人黑色素瘤A875两种黑色素瘤细胞，用RT-PCR、Western blot、基因沉默、染色质免疫共沉淀（ChIP）、激光共聚焦、流式细胞等技术，首先弄清异甘草素如何影响肿瘤细胞糖酵解，明确对糖代谢的影响是否通过靶向HIF-1起作用；接下来阐明糖代谢改变与ROS升高的关系，最后确定异甘草素诱导黑色素瘤细胞凋亡的机制。本研究将为筛选靶向肿瘤能量代谢的天然产物研究提供有益尝试。

中文关键词： 糖酵解；能量代谢；活性氧；异甘草素；HIF

英文摘要： Our previous research has shown that isoliquiritigenin could induce apoptosis in melanoma cells, which accompanied by down regulation of hypoxia-inducible factor (HIF - 1 alpha) mRNA expression, the decrease of content of lactate and glucose intake, and accompanied by increase of ATP content reactive oxygen species (ROS) level; and potential mitochondrial depolarization phenomenon. We hypothesized that isoliquiritigenin may be targeted HIF - 1, lower levels of aerobic glycolysis, raised oxidative phosphorylation metabolic pathway, causing increased formation of ROS, activation of mitochondrial or endoplasmic reticulum stress apoptosis pathway. To verifying our hypothesis, two lines of tumor cells (mouse melanoma B16F0, human melanoma A875) and following technology, such as RT-PCR, Western blot, gene silencing, chromatin immune co-precipitation (ChIP), laser confocal and flow cytometry etc. will be used in this research protocol. First, we will find out whether isoliquiritigenin have effects on the glucose metabolism in melanoma cells, then we will clear and definite whether these effects on glucose metabolism is through targeted HIF - 1, and definite the relationship between glucose metabolism changes and elevated ROS level. Finally, we will determine the mechanism of apoptosis in melanoma cells induced by isoliquiritigenin. For clinical application of such kind of compound laid a foundation, in order to find out HIF-1 inhibitor from natural products for treatment of cancer provided beneficial references, for further development and utilization of isoliquiritigenin and its analogues provide the basis of theory and practice.

英文关键词： Isoliquiritigenin;Energy metabolism;ROS;HIF;glycolysis