核糖体蛋白基因RPLP0对急性髓系白血病细胞增殖和凋亡的调控作用

Effect of ribosomal protein gene RPLP0 on proliferation and apoptosis of acute myeloid leukemia cells

目的研究核糖体蛋白侧柄亚单位P0(ribosomal protein lateral stalk subunit P0,RPLP0)对急性髓系白血病(AML)细胞增殖和凋亡的调控作用。方法用qRT-PCR和Western blot检测健康者(n=15)和AML患者(n=30)骨髓标本、AML细胞系(THP-1)及PBMC细胞中RPLP0的表达。用Lipofectamine 2000分别将RPLP0的短发夹RNA(shRNA-RPLP0组)、shRNA阴性对照(shRNA-NC组)、过表达RPLP0的质粒(pcDNA3.1-RPLP0组)以及对照质粒(pcDNA3.1-NC组)转染到THP-1细胞中,MTT法测定细胞活力,流式细胞术测定细胞凋亡,qRT-PCR和Western blot检测细胞中p53、PTEN、PI3K、p-PI3K、AKT和p-AKT的表达。结果与健康者相比,AML患者骨髓中RPLP0的mRNA和蛋白表达均显著升高(P<0.001)。与PBMC相比,THP-1细胞中RPLP0的mRNA和蛋白表达均显著升高(P<0.001)。与shRNA-NC组相比,shRNA-RPLP0组的相对细胞活力降低(P<0.001),而细胞凋亡率升高(P<0.001)。与shRNA-NC组相比,shRNA-RPLP0组的p53和PTEN蛋白表达量显著升高,而p-PI3K和p-AKT蛋白表达量显著降低(P<0.05)。与pcDNA3.1-NC组相比,pcDNA3.1-RPLP0组相对细胞活力升高(P<0.05),p53和PTEN蛋白表达量降低(P<0.05),p-PI3K和p-AKT蛋白表达量升高(P<0.05)。结论下调RPLP0的表达可抑制AML细胞的增殖并诱导凋亡,RPLP0对AML细胞增殖和凋亡的影响是通过p53-PTEN-PI3K-AKT轴介导的。

Objective To explore the regulation of ribosomal protein gene ribosomal protein lateral stalk subunit P0(RPLP0)on the proliferation and the apoptosis of acute myeloid leukemia(AML)cells.Methods The expression of RPLP0 was detected in bone marrow specimens of 15 healthy controls and 30 AML patients,AML cell lines(THP-1)and PBMC by qRT-PCR and Western blot.RPLP0 short hairpin RNA(shRNA-RPLP0 group),shRNA negative control(shRNA-NC group),RPLP0 overexpression plasmid(pcDNA3.1-RPLP0 group)and control plasmid(pcDNA3.1-NC group)were transfected into THP-1 cells using Lipofectamine 2000,respectively.Cell viability was determined by MTT method,cell apoptosis was determined by flow cytometry,qRT-PCR and Western blot were used to detect the expression of p53,PTEN,PI3K,p-PI3K,AKT and p-AKT in cells.Results The expression levels of RPLP0 mRNA and protein in bone marrow of AML patients were significantly higher than those of healthy controls(P<0.001).Compared with PBMC,the mRNA and protein expression levels of RPLP0 in THP-1 cells were significantly increased(P<0.001).Compared with shRNA-NC group,the relative cell viability decreased in shRNA-RPLP0 group(P<0.001),while the apoptosis rate was increased(P<0.001).Compared with shRNA-NC group,the p53 and PTEN protein expression levels were significantly increased in shRNA-RPLP0 group,while the expression levels of p-PI3K and p-AKT protein were significantly reduced(P<0.05).Compared with pcDNA3.1-NC group,the relative cell viability was increased in pcDNA3.1-RPLP0 group(P<0.05),the expression of p53 and PTEN protein was decreased(P<0.05),and the expression of p-PI3K and p-AKT protein was increased(P<0.05).Conclusion Down-regulating the expression of RPLP0 can inhibit the proliferation of AML cells and induce the apoptosis through the p53-PTEN-PI3K-AKT axis.