增强Mel 18基因表达抑制U937细胞生长

Overexpression of Mel 18 Inhibiting the Proliferation of U937 Cells

目的研究Mel18基因在急性白血病细胞株U937的表达情况及其对U937细胞生长的影响。方法RT-PCR方法检测Mel18在U937细胞中的表达。用亚克隆技术构建真核表达质粒pLenti6/V5-Mel18,脂质体转染法将重组质粒pLenti6/V5-Mel18和对照质粒pLenti6/V5-LacZ分别导入U937细胞,以RT-PCR法和流式细胞仪检测U937细胞转染后Mel18的表达情况,CCK-8法检测细胞增殖活性。结果U937细胞中未检测到Mel18的表达。成功构建了Mel18的正义真核表达质粒pLenti6/V5-Mel18,并将其成功导入U937细胞。pLenti6/V5-Mel18质粒转染U937细胞24h后,流式检测Mel18蛋白的阳性表达率为(23.75±2.32)%。相对于亲代细胞,转染pLenti6/V5-Mel18组与转染pLenti6/V5-LacZ组的24h增殖活性分别为(70.86±1.08)%和(95.89±1.26)%,48h增殖活性为(46.93±1.12)%和(95.43±1.63)%,差异均有极显著性意义(P<0.01)。结论U937细胞不表达Mel18基因,上调Mel18的表达能明显抑制U937细胞的生长。

Objective To explore the expression of Mel 18 gene in the human leukemic cell line U937 and effects of exogenous Mel 18 gene on the proliferation of U937 cells in vitro. Methods RT-PCR was used to detect the expression of Mel 18 in U937 cells. Recombinant eukaryotic expression vector pLenti6/V5-Mel 18 was then constructed by subcloning technique and subsequently transfected into U937 cells by LipofectamineTM 2000. The positive expression rate of Mel 18 was detected by flow cytometry 24 h after vector transfection was performed, while effects of Mel 18 transfection on the growth of U937 cells was investigated via CCK-8. Results No Mel18 expression could be detected in U937 cells. The erukaryotic expression vector pLenti6/V5-Mel 18 was constructed successfully. The positive expression rate of Me118 protein after the pLenti6/V5-Mel 18 transfection was（23.75±2.32）%. Compared with the parental cells, the relative proliferation activity of pLenti6/V5-Mel 18 transfected U937 vs pLenti6/V5-LacZ transfected parterner was（ 70.86±1.08） % vs（95.89±1.26）% at 24 h（ P〈 0.01 ） and（46.93±1.12）% vs（95.43±1.63）%（P〈0.01） at 48 h in vitro respectively, with the difference being significant. Conclusion Mel 18 is not expressed in acute leukemia cell line U937 and up-regulated expression of Mel 18 gene in leukemic cells may suppress their proliferation.