基于青蒿素小分子探针的弥散性大B细胞淋巴瘤细胞铁死亡机理研究

Ferroptosis is a newly discovered iron ion dependent cell death mode, the comprehensive use of molecular biology, cell biology and chemical protein mass spectrometry technology, Ferroptosis mechanism of diffuse large B cell lymphoma cell will be studied based on artemisinin small molecular probes. First of all, MTT assays were performed to study whether artemisinin has an inhibitory effect on diffuse large B cell lymphoma cell proliferation in a cellular level; then, the ferroptosis mechanism of artemisinin inhibitting the proliferation of diffuse large B cell lymphoma cells will be studied by Annexin V/PI double staining and fluorescence microscopy, reactive oxygen species (ROS), lipid peroxide (LPO) test and Caspase-3 activity detection; and then, transcriptome sequencing will be used to study expression of the tumor genes involved with ferroptosis; to study interactions of artemisinin with ferroptosis proteins, chemical protein mass spectrometry will be utilized; finally, activity and mechanism of artemisinin in vivo will be studied using human diffuse tumor large B cell lymphoma cell nude mouse model. Based on the above studies, we will explore the mechanism of artemisinin small molecular probe on the ferroptosis of diffuse large B cell lymphoma cells, it will provide a theoretical basis for the further development of antitumor drugs.

铁死亡是一种新发现的铁离子依赖性的细胞死亡模式，本项目计划综合运用分子生物学、细胞生物学和化学蛋白质谱学等技术，研究青蒿素小分子探针导致弥散性大B细胞淋巴瘤细胞铁死亡的作用机理。我们将利用MTT法检测青蒿素对弥散性大B细胞淋巴瘤细胞增殖的抑制活性；利用Annexin V/PI双染色法和荧光显微镜、活性氧ROS检测法、脂质过氧化物（LPO）测试法、Caspase-3活性检测等研究其导致弥散性大B细胞淋巴瘤细胞铁死亡的作用机理；然后，应用转录组测序和化学蛋白质组技术，研究铁死亡过程中肿瘤细胞基因表达的变化情况,以及其作用蛋白；最后，利用人弥漫性大B细胞淋巴瘤小鼠肿瘤模型，研究青蒿素通过铁死亡机制抑制弥散性大B细胞淋巴瘤细胞增殖的动物体内活性。通过上述研究，我们将探索蒿素小分子探针导致弥散性大B细胞淋巴瘤细胞铁死亡的作用机理，为进一步将其开发成新的抗肿瘤药提供理论基础。

铁死亡是一种新发现的铁离子依赖性的细胞死亡模式。本项目综合运用分子生物学、细胞生物学、转录组测序和化学蛋白质谱学等技术，研究青蒿素类化合物诱导淋巴瘤细胞铁死亡的作用机理。研究发现：1）青蒿素类化合物抑制弥散大B细胞淋巴瘤和Burkitt淋巴瘤细胞增殖，诱导了淋巴瘤细胞的铁死亡；2）ATF4-CHOP-CHAC1通路的激活增强了青蒿琥酯诱导淋巴瘤细胞铁死亡的活性；4）青蒿琥酯通过作用于淋巴瘤细胞VDAC和Xc-系统,诱导了淋巴瘤细胞铁死亡；5）青蒿琥酯可能通过作用于淋巴瘤细胞中的HSP90，抑制了肿瘤细胞增殖；5）采用小鼠移植瘤模型进一步证实了青蒿琥酯在体内能诱导铁死亡抑制淋巴瘤细胞增殖。这些研究结果说明青蒿素类化合物导致的淋巴瘤铁死亡具有重要的研究价值，可以为进一步提高青蒿素类化合物抑制淋巴瘤增殖活性的研究提供新的理论。