双氢青蒿素通过c-Myc/HIF-1α调控肿瘤细胞糖代谢的分子机制

Cancer has become the first killer of human health. To find an effective anti-tumor drug has been the therapeutic problem and research hotspot in cancer treatment. Aerobic glycolysis (Warburg effect) is the most notable difference between cancer cells and normal cells. Our previous studies have found that the dihydroartemisin (DHA) can inhibit tumor growth in vitro. It’s found that tumor glucose metabolism was affected using whole genomic gene chip of cancer cells. This project will evaluate the anti-tumor effect of DHA use of human myeloid leukemia cell K562 model, aims to clarify the main signal to DHA antitumor pathways. After treat with DHA, we will analyze the expression changes of the key regulatory factor of Warburg effect (c-Myc, HIF-1α and p53) and their important enzyme of downstream (HK2, PFK1, PDK1, PKM2 and LDH-A), to explore the relationship of DHA to the Warburg effect and clarify its mechanism of anti-tumor. This project will provide experimental basis for application of DHA in tumor treatment, and provide theoretical support for anti-tumor mechanism of artemisinin derivatives.

恶性肿瘤已经成为人类健康的第一杀手，寻找有效的肿瘤治疗药物一直是肿瘤研究领域的热点和难点。有氧糖酵解（Warburg效应）是肿瘤细胞代谢区别于正常细胞的最显著特征。课题组前期研究证实双氢青蒿素（DHA）具有明显的抗肿瘤作用；通过全基因组表达谱芯片结合生物信息学方法分析发现，DHA能影响肿瘤糖代谢，但其具体机制尚不明确。本项目重点研究Warburg效应中关键因子c-Myc、HIF-1α和p53，通过RT-PCR、Western Blot、激光共聚焦等方法检测DHA作用不同时间后人髓系白血病K562细胞c-Myc、HIF-1α和p53及其下游关键酶（HK2、PFK1、PDK1、PKM2和LDH-A）的表达变化，探索DHA对糖代谢的调控作用，明确其抗肿瘤作用机制。本项目通过探索DHA的抗肿瘤作用机制，将为DHA及青蒿素类衍生物应用于肿瘤治疗提供科学依据，具有重要的理论意义和实际应用价值。

肿瘤治疗是关系到国计民生的重大医疗难题，寻找有效的肿瘤治疗药物一直是肿瘤研究领域的热点和难点。有氧糖酵解（Warburg效应）是肿瘤细胞代谢区别于正常细胞的最显著特征。双氢青蒿素（DHA）作为青蒿素类衍生物的代表，具有影响肿瘤糖代谢的功能。本项目利用人髓系白血病K562细胞模型研究DHA对肿瘤细胞的杀伤作用机制，旨在阐明DHA影响肿瘤细胞糖酵解作用的关键靶点。通过RT-PCR、Western Blot、免疫荧光等方法检测DHA作用不同时间后Warburg效应中关键因子c-myc、HIF-1α和p53及糖代谢过程中其下游的关键蛋白（GLUT1，HK2、PFK1、PDK1、PKM2和LDH-A）的表达变化，探索DHA对肿瘤细胞能量代谢的调控作用，明确其作用机制。通过本项目的实施，我们发现DHA通过下调PKM2和GLUT1的表达而发挥抑制肿瘤细胞糖酵解作用，给予DHA后，PKM2和GLUT1的表达无论在基因水平还是在蛋白水平均受到抑制。由此，我们认为PKM2和GLUT1为DHA影响白血病细胞糖酵解的关键靶点。本项目将为青蒿素类衍生物应用于肿瘤治疗提供实验依据，为阐明青蒿素类衍生物抗肿瘤机制研究提供支撑，具有重要的理论意义和实际应用价值。