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目的:研究人黑素瘤分化相关基因-7(melanoma differentiation associated gene-7,mda-7;又称IL-24)及其剪接体IL-24 delE5与急性髓系白血病(acute myeloid leukemia,AML)细胞分化的关系。方法:十四烷酰佛波醇-乙酯(12-O-tetrade-canoylphorbol-13-acetate,TPA)处理人急性髓系白血病细胞系U937、HL-60,real-time PCR及Western blotting检测细胞中mda-7/IL-24和IL-24 delE5的表达,FACS检测细胞表面CD11b、CD14和CD115的表达。siRNA干扰U937、HL-60细胞中mda-7/IL-24和IL-24 delE5的表达后,再以TPA诱导细胞分化,FACS检测CD11b、CD14和CD115的表达。用前期构建的mda-7/IL-24和IL-24 delE5载体转染U937、HL-60及AML-M5患者的原代白血病细胞,观察异位过表达mda-7/IL-24和IL-24 delE5能否诱导白血病细胞向单核细胞分化。结果:TPA在诱导U937、HL-60细胞向单核细胞分化过程中显著促进mda-7/IL-24及其剪接体IL-24 delE5的表达,用siRNA干扰mda-7/IL-24及IL-24 delE5的表达后,TPA诱导U937、HL-60细胞向单核细胞分化的作用也被阻断。异位过表达mda-7/IL-24和IL-24 delE5可诱导U937、HL-60以及原代AML细胞向单核细胞分化:U937、HL-60细胞表面CD11b和CD14表达均显著升高,CD115表达仅在U937细胞有显著升高;3例AML-M5患者的原代AML细胞中CD11b、CD14及CD115表达均有不同程度升高,细胞呈现单核细胞的形态特征。结论:TPA可通过上调mda-7/IL-24及其剪接体IL-24 delE5的表达诱导白血病细胞向单核细胞分化。
Objective: To study the expression of melanoma differentiation associated gene-7 (mda-7; IL-24) and its splice variant IL-24 delE5 and acute myeloid leukemia (AML) Relationship between cell differentiation. Methods: Human acute myeloid leukemia cell line U937 was treated with 12-O-tetrade-canoylphorbol-13-acetate (TPA), HL-60, real-time PCR and Western blotting The expression of mda-7 / IL-24 and IL-24 delE5 was detected by FACS. The expression of CD11b, CD14 and CD115 on the cell surface was detected by FACS. After interfering with the expression of mda-7 / IL-24 and IL-24 delE5 in U937 and HL-60 cells, siRNA was used to induce cell differentiation by TPA. The expression of CD11b, CD14 and CD115 was detected by FACS. The primary leukemic cells of U937, HL-60 and AML-M5 were transfected with the mda-7 / IL-24 and IL-24 delE5 vectors and the expression of mda-7 / IL- 24 delE5 can induce leukemia cells to monocyte differentiation. Results: TPA could significantly promote the expression of mda-7 / IL-24 and its spliceosome IL-24 delE5 in the process of inducing U937 and HL-60 cells to monocyte differentiation. Interfering mda-7 / IL-24 and IL -24 delE5 expression, TPA-induced U937, HL-60 cells to monocytes differentiation is also blocked. Ectopic overexpression of mda-7 / IL-24 and IL-24 delE5 induced differentiation of U937, HL-60 and primary AML cells into monocytes: the expression of CD11b and CD14 on U937 and HL-60 cells were significantly increased, The expression of CD115 in U937 cells was significantly increased. The expression of CD11b, CD14 and CD115 in 3 AML-M5 primary AML cells increased to some extent. The morphology of monocytes was characterized by mononuclear cells. CONCLUSION: TPA induces the differentiation of leukemia cells into monocytes by up-regulating the expression of mda-7 / IL-24 and its spliceosome IL-24 delE5.