RBM38调节FZD1 mRNA稳定性介导急性髓系白血病细胞HL-60的增殖

RBM38 Mediates the Proliferation of Acute Myeloid Leukemia Cells HL-60 by Regulating FZD1 mRNA Stability

目的:研究RNA结合基序蛋白38(RBM38)对人急性髓系白血病细胞株HL-60增殖的调控功能和作用机制。方法:分别构建过表达、敲低RBM38的慢病毒载体,使用慢病毒感染HL-60细胞后,Western blot方法检测细胞内RBM38的表达水平,分别用CCK-8法和PI染色结合流式细胞术检测HL-60细胞的增殖和周期,通过RNA免疫共沉淀结合实时定量PCR方法检测RBM38与靶mRNA的结合情况,放线菌素D处理结合实时定量PCR方法检测RBM38对靶mRNA稳定性的影响。结果:通过慢病毒介导的基因转移可成功实现HL-60细胞内RBM38的过表达或表达抑制。过表达RBM38可显著促进HL-60细胞的增殖活性和细胞周期运行;相反,抑制内源性RBM38的表达可抑制HL-60细胞增殖和细胞周期。RBM38可与FZD1 mRNA结合,促进其mRNA的稳定。结论:RBM38通过促进FZD1 mRNA的稳定调节HL-60细胞的增殖。

Objective: To explore the regulatory function of RNA binding motif protein 38(RBM38) in human acute myeloid leukemia cells HL-60 and its mechanism. Methods: The lentivirus carriers of overexpressed and knockdown RBM38 were constructed. After HL-60 cells were transfected, Western blot was used to analyze the expression level of RBM38 in HL-60 cells. The cell proliferation and cycle of HL-60 were detected by CCK-8 assay and flow cytometry assay, respectively. RNA immunoprecipitation coupled real-time PCR(RIP-qPCR) was used to detect the combination of RBM38 with mRNAs. Actinomycin D treatment followed by real-time PCR(AcD-qPCR) was used to detect the effect of RBM38 on the stability of target mRNAs. Results: RBM38 in HL-60 cells was overexpressed or inhibited by lentivirus transduction. Overexpressed RBM38 promoted the cell cycle and proliferation of HL-60, while RBM38 knockdown repressed the two processes. RBM38 showed an interaction with FZD1 mRNA and enhancement of its stability.Conclusion: RBM38 can regulate cell proliferation of HL-60 by improving the stability of FZD1 mRNA.