Twist-1通过MPL促进髓系白血病细胞增殖和耐药

发布时间：2018-09-06 19:33

【摘要】：目的:检测Twist-1与骨髓增生性白血病病毒癌基因(myeloproliferative leukemia virus oncogene,MPL)在髓系白血病患者和髓系造血系统恶性肿瘤细胞系中表达的相关性,并探讨Twist-1是否通过MPL对髓系白血病细胞增殖、耐药发挥促进作用。方法:选取中国医学科学院血液病医院2005年1月至2008年12月初次诊断为急性髓系白血病(acute myeloid leukemia,AML)、慢性粒细胞白血病(chronic myeloid leukemia,CML)患者的骨髓标本41例(其中AML 23例、CML 18例),用Real-time PCR检测AML、CML患者以及髓系造血系统恶性肿瘤细胞系中Twist-1和MPL mRNA的表达情况,并分析其相关性。构建MPL过表达载体,制备慢病毒并感染髓系白血病细胞系K562、U937,通过细胞计数实验、集落形成实验以及药物敏感实验评价其对白血病细胞增殖、集落形成能力以及耐药的影响,并进一步确定Twist-1是否通过MPL发挥白血病促进作用。结果:在U937和K562细胞中过表达Twist-1明显增加MPL蛋白水平(P0.05),敲降Twist-1后MPL mRNA的表达水平明显下降(P0.01);AML、CML患者骨髓单核细胞(bone marrow mononuclear cells,BMMCs)中Twist-1 mRNA表达与MPL mRNA表达水平呈显著正相关(P0.05)。过表达MPL使K562和U937细胞对化疗药物柔红霉素及伊马替尼的敏感性显著降低(P0.01),且提高K562-MPL、U937-MPL的细胞增殖、集落形成数目(均P0.01);干扰Twist-1并过表达MPL显著降低髓系白血病细胞增殖和集落形成(均P0.01)。结论:Twist-1通过MPL促进AML、CML白血病细胞的增殖、存活和耐药。
[Abstract]:Objective: to investigate the expression of Twist-1 and bone marrow proliferative leukemia virus oncogene (myeloproliferative leukemia virus oncogene,MPL) in myeloid leukemia patients and myeloid hematopoietic malignant tumor cell lines, and to explore whether Twist-1 can promote the proliferation of myeloid leukemia cells through MPL. Drug resistance plays a catalytic role. Methods: from January 2005 to December 2008, 41 bone marrow specimens from patients with acute myeloid leukemia (acute myeloid leukemia,AML) and chronic myelogenous leukemia (chronic myeloid leukemia,CML) were collected from Hematological Hospital of the Chinese Academy of Medical Sciences (AML 23 cases). To detect the expression of Twist-1 and MPL mRNA in AML,CML patients and myeloid hematopoietic malignant tumor cell lines by Real-time PCR. And analyze its correlation. MPL overexpression vector was constructed to prepare lentivirus and infect myeloid leukemia cell line K562U937. The effects of lentivirus on the proliferation, colony forming ability and drug resistance of leukemia cells were evaluated by cell count assay, colony forming assay and drug sensitivity test. And further determine whether Twist-1 plays a role in promoting leukemia through MPL. Results: overexpression of Twist-1 in U937 and K562 cells significantly increased the level of MPL protein (P0.05), and the expression level of MPL mRNA decreased significantly (P0.01) in the bone marrow monocytes of U937 and K562 cells (P0.01). There was a significant positive correlation between Twist-1 mRNA expression and MPL mRNA expression level in the bone marrow monocytes of U937 and K562 cells (P0.05). Overexpression of MPL significantly decreased the sensitivity of K562 and U937 cells to the chemotherapeutic drugs daunorubicin and imatinib (P0.01), and increased the proliferation of K562-MPLU37-MPL cells. The number of colony formation (all P0.01), interfering with Twist-1 and overexpression of MPL significantly decreased the proliferation and colony formation of myeloid leukemia cells (P0.01). Conclusion: Twist-1 promotes the proliferation, survival and drug resistance of AML,CML leukemia cells through MPL.
【作者单位】：中国医学科学院血液病医院血液学研究所实验血液学国家重点实验室;
【基金】：国家自然科学基金资助项目(No.81670158,No.81470278,No.81600138) 天津市应用基础与前沿技术研究计划资助项目(青年项目)(No.15JCQNJC10300) 协和青年科研基金滚动资助项目(No.33320140066)~~
【分类号】：R733.7

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