DNMT3A突变导致SLC7A11高表达介导AML耐药的机制研究

DNMT3A mutation, as an important molecular genetic change in acute myeloid leukemia (AML), can persist steadily during the initial and recurrent stages of the disease and is closely related to drug resistance and recurrence of AML. However, the mechanism is still unclear. High expression of SLC7A11，which is closely related to the redox balance in tumor cells so that to affect the metabolism of chemotherapeutic drugs, tumor growth and migration, has not been reported in AML. Based on our previous work, we speculate that the DNMT3A mutation may increase GSH synthesis in AML cells by upregulating SLC7A11 expression, thereby affecting the process of cell proliferation and drug resistance. In this project, we used gene overexpression and gene knockout techniques with DNMT3A R882H mutation cell line to clarify the role of SLC7A11 gene in AML cells. Then we applied expression microarray and bioinformatics analysis to explore the effect of SLC7A11 on the expression of AML related drug resistance genes and NOD/SCID mouse model was established for in vitro validation and analysis. Ultimately, SLC7A11 specific inhibitor was used for exploring the possibility of SLC7A11 as a new therapeutic target of AML, which could provide a new clinical idea for the targeted treatment of AML.

DNMT3A突变作为急性髓系白血病（AML）中一种重要的分子遗传学改变，可在疾病的初始至复发阶段持续稳定存在，与AML的耐药和复发有密切关系，但相关机制仍不明确。SLC7A11高表达与肿瘤细胞内氧化还原平衡密切相关，可影响化疗药物代谢、肿瘤生长及迁徙等，但在AML中尚未见报道。基于我们的前期工作，我们推测：DNMT3A突变可能通过上调SLC7A11表达，使AML细胞内GSH合成增加，进而影响细胞的增殖及出现耐药。在本项目中，我们利用DNMT3A R882H突变的细胞系，使用基因过表达和基因敲除技术，明确SLC7A11基因在AML细胞中的作用，再通过表达谱芯片、生物信息学分析发掘SLC7A11表达改变后对AML相关耐药基因表达的影响，并构建NOD/SCID小鼠模型进行体外验证分析。同时使用SLC7A11特异性抑制剂研究其作为AML新的治疗靶点的可能性，为AML的靶向治疗提供新的临床思路。

DNMT3A突变作为急性髓系白血病（AML）中一种重要的分子遗传学改变，可在疾病的初始至复发阶段持续稳定存在，与AML的耐药和复发有密切关系，但相关机制仍不明确。SLC7A11高表达与肿瘤细胞内氧化还原平衡密切相关，可影响化疗药物代谢、肿瘤生长及迁徙等，但在AML中尚未见报道。我们以前期的DNMT3A R882H突变细胞系为基础，通过基因过表达和基因敲除技术构建SLC7A11过表达细胞株和SLC7A11-/-细胞株，进行相应生物行为学检测提示两种细胞株在细胞形态及免疫表型上没有明显差异，但SLC7A11过表达细胞株增值能力更强且具有更强的抗凋亡能力。后期我们将继续进行表达谱芯片检测，发掘SLC7A11表达改变后对AML相关耐药基因表达的影响，并构建NOD/SCID小鼠模型进行体外验证分析。