低极性人参皂苷安留克抑制单核细胞白血病髓外浸润的机制研究

Acute monocytic leukemia with high incidence of extramedullary infiltration and poor prognosis, there is no specific effective medicine. In our previously study, low polarity of panax ginsenosides an-liu-ke (ALK) was screened and identified by biological activity assay of anti-leukemia as the index. The invention patent of ALK applied has been authorized. The specific elements and content of ALK was clear and definite, mainly composed of five monomers F4, Rh4 and S-Rg3, etc. Our study also showed that ALK was significantly effective not only to suppress proliferation in monocytic leukemic patients’ primary leukemic cells and monocytic SHI-1 cell line, but also to resist against their invasion and infiltration of brain tissue and the other organs in model mice. The mechanism of anti-extracellular infiltration of ALK may be involved in the regulation of multiple target genes. Therefore, this project is to observe effective of ALK on suppression proliferation, migration and infiltration by using monocytic leukemic patient’s primary leukemic cells and SHI-1 leukemic cells, meanwhile, establishing three type models with monocytic leukemic patient’s primary leukemic cells and SHI-1 leukemic cells, as well as detecting panax ginsenosides monomers of ALK in brain tissue of model mice. The differences of candidate target genes of leukemic cells induced by ALK treatment will be screened using new technology of digital gene expression profiling, the candidate target genes were confirmed in vitro and vivo of mouse models by using multiple methods, and intracellular signal pathways mediated by ALK will be constructed. The preliminary analysis of correlation will be made among experimental results, extramedullary infiltration and curative effect in clinical patients, it may possess reference value for predicting the extramedullary infiltration and improving prognosis in monocytic leukemia patients, also provide the experimental evidence to develop ALK as a new Chinese patent drug for effectively treating extramedullary infiltration of leukemic patients without obvious side efficacy.

髓外浸润是急性单核细胞白血病的严重并发症，预后差，尚无特效药物。前期工作以生物学活性为考察指标，筛选并确定低极性人参皂苷安留克（ALK）抗白血病作用明显，发明专利被授权。成分含量明确主含F4、Rh4、S-Rg3等5个人参皂苷单体，抑制单核白血病患者原代及SHI-1细胞增殖及其在小鼠模型脑组织和其它脏器的浸润。设想ALK抗单核白血病髓外浸润机制可涉及到多重靶基因的调控，本课题采用单核白血病患者原代和SHI-1细胞株，及髓外浸润的三种小鼠模型，观察ALK体内外抑制单核白血病细胞增殖、迁移和浸润作用，检测模型小鼠脑组织中ALK的单体。数字基因表达谱新技术筛选ALK作用于白血病髓外浸润差异表达的基因，多种方法验证后确定作用的关键靶基因，并构建相关信号通路，初步分析与患者临床髓外浸润及疗效的相关性，对今后预测白血病患者髓外浸润及改善预后有参考价值，为研发安全有效抗髓外浸润的中药新药提供实验依据。

髓外浸润是急性单核细胞白血病的严重并发症，预后差，尚无特效药物。前期工作以生物学活性为考察指标，筛选并确定低极性人参皂苷安留克（ALK）抗白血病作用明显，成分含量明确，发明专利被授权。. 通过本课题实施，获得有价值的结果：①证明ALK体内外有效地抑制单核白血病细胞增殖、迁移和浸润。②数字基因表达谱筛选出ALK诱导差异表达的基因，与白血病的发生发展密切相关。③有效地治疗SHI-1单核白血病髓外浸润小鼠模型，抑制白血病细胞在体内增殖及中枢神经系统浸润。④采用患者原代粒-单核白血病细胞输入，成功制备白血病PDX小鼠模型，ALK有效地抑制原代白血病细胞在小鼠体内增殖及浸润。⑤原代白血病细胞对ALK敏感且下调迁移浸润相关基因的患者，临床疗效较好，获得完全缓解，提示对预测疗效有一定的参考价值。⑥质谱分析ALK治疗组小鼠脑组织检测出其主含成分人参皂苷Rd、S-Rg3和F4单体，提示ALK可通过血脑屏障而抗白血病脑浸润。⑦计划外增加的工作：从ALK分离出所含单体，其中人参皂苷F4、S-Rg3单体具有抑制白血病细胞增殖、诱导分化及抗迁移浸润作用。⑧发表相关论文8篇，其中SCI收录6篇，第一标注基金批准号7篇，第二标注1篇。培养博士和硕士生5名，晋升副高职称1名，学术会议报告7次。.科学意义及应用前景：①揭示ALK抗单核白血病髓外浸润的作用及机制，为研发安全有效抗白血病的中药新药/辅助药物提供实验依据。②成功制备人原代粒-单核白血病髓外浸润PDX小鼠模型，符合患者白血病的特征，能够较客观准确地反映ALK抗白血病的疗效。③患者原代白血病细胞对ALK敏感，且下调迁移浸润相关基因表达，其临床疗效较好，完全缓解率高，提示对预测疗效有一定的参考价值。④ALK所含单体能够透过血脑屏障，设想将来用ALK治疗白血病脑浸润，具有服用方便，无需鞘内注射等优点。