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Larp4b是人类LARP家族中的一员,为广谱翻译因子,可结合RNA并促进mRNA的翻译。前期本实验室的研究工作提示白血病患者存在Larp4b基因表达变化。本研究检测Larp4b基因在造血各系细胞中的表达量,敲降Larp4b表达对Lin-造血细胞生物行为的影响。利用RT-PCR检测Larp4b基因在造血各系细胞中的表达,用病毒上清感染Lin-细胞,分选EGFP+细胞,PI染色法检测细胞周期,AnnexinV-PE/7-AAD双染法检测细胞凋亡,并进行集落形成实验(colony forming cell assay,CFC)。结果表明:Larp4b基因在造血各系细胞中表达不同,Larp4b基因在粒系和巨噬祖细胞(GMP)以及髓细胞(M)的表达较高,在T淋巴细胞中表达明显低,抑制Larp4b表达不影响Lin-造血细胞体外集落形成能力,不影响Lin-造血细胞的细胞周期和凋亡。结论:Larp4b基因在髓系细胞中表达较高。抑制Larp4b表达不影响Lin-造血细胞集落形成能力,不影响Lin-造血细胞的增殖,这为后续Larp4b体内实验研究奠定了基础。
Larp4b, a member of the human LARP family, is a broad-spectrum translation factor that binds RNA and facilitates the translation of mRNA. Preliminary research work in this laboratory prompted the presence of Larp4b gene expression in patients with leukemia. This study was to investigate the effect of Larp4b knockdown on the biological behavior of Lin-hematopoietic cells by detecting the expression of Larp4b in hematopoietic lineage cells. The expression of Larp4b gene in hematopoietic lineage cells was detected by RT-PCR. Lin-cells were infected with virus supernatant and EGFP + cells were sorted. PI staining was used to detect the cell cycle. Cell apoptosis was detected by Annexin V-PE / 7-AAD double staining The colony forming cell assay (CFC) was performed. The results showed that the Larp4b gene was expressed differently in hematopoietic lineage cells. The Larp4b gene was highly expressed in myeloid and macrophage progenitor cells (GMP) and myeloid cells (M), and was significantly lower in T lymphocytes and inhibited Larp4b expression Does not affect Lin-hematopoietic cells in vitro colony-forming ability, does not affect the Lin-hematopoietic cell cycle and apoptosis. Conclusion: The Larp4b gene is highly expressed in myeloid cells. Inhibition of Larp4b expression did not affect the colony forming ability of Lin-hematopoietic cells, and did not affect the proliferation of Lin-hematopoietic cells, which lays a foundation for the subsequent study of Larp4b in vivo.