基于“遗邪内伏”理论研究中药干预染色体不稳定性调节CDK2泛素化通路诱导AML细胞分化作用机制

染色体不稳定性（CIN）是导致基因突变、抑癌基因缺失，引发急性髓细胞白血病（AML）发生、发展的关键因素。AML重要特征是细胞分化障碍，诱导分化是主要治疗方法。干预CIN能够影响AML细胞的增殖和分化。CDK2蛋白在AML细胞分化中发挥关键作用，可作为一个潜在治疗靶点。中医理论“遗邪内伏”是AML重要病机，虚实并治为主要治则，前期临床研究发现：以雄黄、青黛配伍酒萸肉为中药组方，可诱导细胞分化，改善患者生存期，但作用机制不清。本课题以AML小鼠及细胞株为研究对象，探索中药干预CIN诱导细胞分化的生物学特征，检测CDK2泛素化水平及KLHL6、PRDX2、MAFB细胞分化关键蛋白的表达；揭示中药干预CIN诱导AML细胞分化的作用机制。基于“遗邪内伏”理论探讨中药治疗AML的组方用药规律和诱导细胞分化作用机制，对于阻止疾病复发、延长白血病缓解期具有重要研究价值。

Chromosomal instability (CIN) is the key factor that leads to gene mutation, tumor suppressor gene deletion and AML occurrence and development. Cell differentiation disorder is an important feature of AML, and induced differentiation is an important treatment. CDK2 protein plays an important role in the process of AML cell differentiation and can be a potential new target for AML treatment. The TCM theory of “Endogenous pathogenic factors” is an important pathogenesis of AML. The combination of deficiency and excess is the main treatment principle. In the early clinical study, realgar indigo Cornus officinalis can improve the survival time of patients, induce cell differentiation, but the mechanism is not clear. The preliminary study of the research group found that the intervention of CIN can affect the proliferation and differentiation of AML cells.In this study, the AML mouse model and cell line were used to detect the effective targets and mechanism of realgar indigo Cornus officinalis in the treatment of AML on cell differentiation induced by CIN. The level of CDK2 ubiquitination , the binding of CDK2 and ubiquitin E3 ligase klhl6 protein are detected by IP Co-IP technology,respectively.The expression of CDK2, KLHL6, Prdx2 and MafB which are cell differentiation key proteins are detected by RT-PCR and Western blot .The purpose of this study is to reveal the mechanism of on the differentiation of AML cells induced by interference of chromosomal instability. Based on the theory of "endogenous pathogenic factors", it is of great research value to explore the regularity and mechanism of traditional Chinese medicine in the treatment of leukemia, induce cell differentiation, prevent disease recurrence and prolong the remission period of leukemia.