摘要
目的探讨细胞质中核仁磷酸蛋白A型突变体(NPM1 mutant A,NPM1 m A)与AKT的相互作用,观察NPM1 m A联合AKT对白血病细胞增殖的影响。方法免疫共沉淀实验观察胞质蛋白NPM1 m A与AKT的相互作用。通过RNA干扰技术抑制NPM1突变阳性的OCI/AML3白血病细胞株中NPM1突变蛋白的表达。不同浓度AKT抑制剂(AKT INHIBITORⅣ,AKTⅣ)处理白血病细胞抑制磷酸化AKT蛋白表达。CCK-8法检测细胞体外增殖能力。结果胞质NPM1 m A能够与内源性的AKT蛋白相互结合。干扰NPM1后,OCI/AML3细胞中NPM1 m A和野生NPM1蛋白表达明显下降(P<0.05),白血病细胞的体外增殖能力亦明显降低(P<0.05)。AKTⅣ处理后,白血病细胞体外增殖较未处理细胞显著下降(P<0.05)。若NPM1沉默合并AKTⅣ处理,则OCI/AML3细胞干扰组体外增殖明显降低(P<0.05)。结论在NPM1突变的白血病细胞中,胞质NPM1 m A能够与AKT相互结合,并参与调控白血病细胞的增殖。
Objective To observe the interaction between nucleophosmin mutant A（ NPM1 m A） and AKT in the cytoplasm,and to investigate the effect of NPM1 m A and AKT on the proliferation of acute myeloid leukemia（ AML） cells. Methods Co-immunoprecipitation assay was used to explore the interaction between NPM1 m A and AKT. The expression of NPM1 and NPM1 m A in OCI / AML3 cells was knocked down by RNA interference. The expression of p-AKT was suppressed by AKT inhibitor Ⅳ（ AKT Ⅳ）. Cell proliferation capability was determined by CCK-8 assay. Results NPM1 m A interacted with AKT in OCI / AML3 cells.Transfection with the targeting plasmids significantly down-regulated the expression of NPM1 m A and NPM1 proteins in OCI / AML3 cells（ P〈0. 05）. The cell proliferation capability was significantly suppressed in the leukemia cells with NPM1 interference（ P〈0. 05）. After treated with AKT Ⅳ, the cell growth was significantly inhibited（ P〈0. 05）. Combination of knocking-down NPM1 and treating with AKT IV significantly inhibited the cell growth of the OCI / AML3 cells with NPM1 interference（ P〈0. 05）. Conclusion Cytoplasmic NPM1 m A interacts with AKT and regulates the proliferation of AML cells.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2014年第22期2267-2271,共5页
Journal of Third Military Medical University
基金
国家自然科学基金面上项目(81271913)
重庆市科委自然科学基金(CSTC2013jcyj A10035)~~
