摘要
目的探究长链非编码RNA(LncRNAs)HOTAIR在急性髓细胞白血病(AML)中的表达,以及AML细胞对柔红霉素(DNR)耐药的分子机制。方法qRT-PCR检测AML细胞株、AML患者骨髓标本、对照组骨髓标本HOTAIR的表达水平。选取U937细胞和THP1细胞为研究对象,构建HOTAIR基因的shRNA载体,分别转染两株细胞,qRT-PCR检测HOTAIR基因表达水平,Western blot检测P-糖蛋白的表达。DNR(0.1μmol/L)处理细胞株,CCK-8试验检测细胞增殖,流式细胞术检测细胞凋亡。结果AML细胞株与AML患者骨髓中,HOTAIR的表达水平明显高于对照组;成功下调HOTAIR在U937细胞和THP1细胞中表达,P-糖蛋白水平随之降低,成功下调HOTAIR的U937细胞和THP1细胞联合DNR有利于抑制细胞增殖、促进细胞凋亡。结论AML中,HOTAIR的异常表达参与对DNR的耐药机制形成。
Objective To investigate the expression of long noncoding RNA(LncRNAs)HOTAIR in acute myeloid leukemia(AML)and the function and molecular mechanism of resistance to daunorubicin(DNR)in AML cells.Methods The qRT-PCR was used to detect leukemia cell lines,primary acute myeloid leukemia cells,the normal expression level of control cells HOTAIR.U937 cells and THP1 cells were selected as research objects.The sh-NC plasmid and sh-HOTAIR plasmid of hairpin RNA(shRNA)were constructed and transfected into these two cells,respectively.The expression of HOTAIR in the cells was detected by qRT-PCR.P-glycoprotein(P-gp)expression in transfected cells was detected by Western blot,DNR(0.1μmol/L)was applied to each cell,CCK8 assay was used to detect cell proliferation,and flow cytometry was used to detect cell apoptosis.Results The expression of HOTAIR in the normal control group was significantly lower than that in the leukemia cell line and the acute myeloid leukemia primary cells;The expression of HOTAIR in U937 cells and THP1 cells was successfully down-regulated,and the level of P-glycoprotein was also decreased.The expression of HOTAIR in U937 cells and THP1 cells was successfully down-regulated,combined with DNR was more conducive to inhibit cell proliferation and promote apoptosis.Conclusion The abnormal expression of HOTAIR in AML is involved in the formation of resistance to DNR.
作者
陈玲琳
熊术道
高申孟
Chen Linglin;Xiong Shudao;Gao Shenmeng(Dept of Hematology,The Second Affiliaed Hospital of Anhui Medial University,Hlefei 230601;Dept of Internal Medicine,The First Hospital of Wenzhou Medical University,Wenzhou 325000)
出处
《安徽医科大学学报》
CAS
北大核心
2020年第11期1676-1680,共5页
Acta Universitatis Medicinalis Anhui
基金
浙江省自然科学基金(编号:LY19H080001)。
