UCA1在AML患者骨髓组织单个核细胞中的表达及对AML细胞增殖凋亡调节作用观察

Expression of UCA1 in bone marrow mononuclear cells of patients with AML and its regulatory effects on proliferation and apoptosis of AML cells

目的观察长链非编码RNA(long non-coding RNAs,LncRNA)尿路上皮癌胚抗原(urothelial carcinoma associated gene 1,UCA1)在急性髓系白血病(AML)患者骨髓组织单个核细胞中的表达情况及对人AML不同亚型细胞株U937、HL60增殖凋亡的调节作用,并探讨其可能作用机制。方法(1)30例AML患者、30例缺铁性贫血患者的骨髓标本,采用RT-PCR检测骨髓组织单个核细胞UCA1。(2)将U937及HL60细胞各分为1、2、3组。1、2组分别用UCA1-siRNA(可抑制UCA1表达)和空载siRNA转染,3组为空白对照(不作任何处理)。分别于转染24、48、72 h取各组细胞,MTT法测算细胞增殖活力。转染48 h时采用流式细胞术观察各组细胞凋亡情况。转染48 h时采用WESTERN Blotting法检测各组细胞Wnt、β-连环蛋白(β-catenin)、p-gsk-3?蛋白。结果AML及缺铁性贫血患者骨髓组织单个核细胞UCA1相对表达量分别为4.96±1.51、1.09±0.93,二者比较,P<0.05。与2、3组比较,转染24、48、72 h时U937及HL-601组细胞增殖活力低(P均<0.05)。U937细胞1、2、3组细胞凋亡率分别为34.20%±1.21%、7.21%±1.41%、6.84%±1.53%,HL-60细胞1、2、3组细胞凋亡率分别为34.21%±1.83%、7.20%±1.56%、7.85%±1.93%,与2、3组比较,U937及HL-60细胞1组细胞凋亡率高(P均<0.05)。与2、3组比较,U937及HL-601组细胞Wnt、β-catenin相对表达量降低,p-gsk-3?蛋白相对表达量升高(P均<0.05)。结论与缺铁性贫血患者比较,AML患者骨髓组织单个核细胞UCA1表达升高。抑制UCA1表达可抑制U937及HL-60细胞的增殖,促进细胞凋亡,其机制可能与UCA1抑制细胞Wnt/β-catenin信号通路有关。

Objective To observe the expression of long non-coding RNA(LncRNA)urothelial carcinoma associat⁃ed gene 1(UCA1)in bone marrow mononuclear cells of patients with acute myeloid leukemia(AML)and its regulatory ef⁃fects on the proliferation and apoptosis of U937 and HL60 cells,and to investigate the possible mechanism of action.Methods①Bone marrow samples from 30 AML patients and 30 iron deficiency anemia patients were examined for UCA1 by RT-PCR.②U937 and HL60 cells were divided into the groups 1,2,and 3,respectively.Cells in the group 1 and group 2 were transfected with UCA1-siRNA(which can inhibit the expression of UCA1)and no-siRNA,respectively,and cells in the group 3 were blank controls(without any treatment).The cells were harvested at 24,48 and 72 h after transfection,and the proliferation activity was measured by MTT assay.At 48 h after transfection,apoptosis was observed by flow cytometry.Wnt,β-catenin,and p-gsk-3ßproteins were detected by Western blotting at 48 h after transfection.Results The relative expression levels of UCA1 were 4.96±1.51 in AML patients and 1.09±0.93 in iron deficiency ane⁃mia patients,respectively,with statistically significant difference(P<0.05).At 24,48 and 72 h after transfection,the proliferation activity of U937 and HL-601 cells was lower in the group 1 than in the groups 2 and 3(P<0.05).The apoptosis rates of U937 cells in the groups 1,2,and 3 were 34.20%±1.21%,7.21%±1.41%,and 6.84%±1.53%;the apop⁃tosis rates of Hl-60 cells in the groups 1,2,and 3 were 34.21%±1.83%,7.20%±1.56%,and 7.85%±1.93%,respec⁃tively.Compared with groups 2 and 3,the apoptosis rates of U937 and Hl-60 cells in the group 1 were higher(both P<0.05).Compared with groups 2 and 3,the relative expression levels of Wnt andβ-catenin in U937 and HL-601 cells de⁃creased,while the relative expression of p-gsk-3ßprotein increased in the group 1(all P<0.05).Conclusion UCA1 expression in bone marrow mononuclear cells is elevated in AML patients in comparison with that of iron deficiency anemia patients.Inhibition of UCA1 expression can inhibit the proliferation and promote apoptosis of U937 and Hl-60 cells,which may be related to the inhibition of Wnt/β-catenin signaling pathway by UCA1.