长链非编码RNA对急性髓系白血病细胞增殖和凋亡调控研究

LncRNA regulates cell proliferation and apoptosis in acute myeloid leukemia

目的长链非编码RNA(LncRNA)在肿瘤的发生发展中起重要的作用,但它们在急性白血病发病和预后中有何功能,仍不清楚。本研究探索LncRNA-ENST00000414355对急性髓系白血病(acute myeloid leukemia,AML)细胞增殖和细胞凋亡的调控作用,并初步探索LncRNA-ENST00000414355和急性白血病临床诊断的关系。方法采用RAN干扰技术对AML细胞系HL-60、K562、THP-1和U937的LncRNA-ENST00000414355进行表达沉默,用MTT法检测细胞增殖,用流式细胞术检测细胞凋亡。收集2016-01-01-2016-10-30广州市第一人民医院初发未治的10例AML患者和5名正常人的骨髓标本及相关临床资料,用实时荧光定量PCR(qPCR)检测LncRNA-ENST00000414355在骨髓中的表达。结果 4种白血病细胞系分别被3条si-RNA干扰后,LncRNA-ENST00000414355表达均明显下调。MTT法检测结果显示,RNA干扰72h后,LncRNA-ENST00000414355干扰细胞组HL-60、K562、THP-1和U937细胞的A值分别为0.706±0.04、0.606±0.005、0.617±0.003和0.808±0.016 6,明显低于阴性对照组的1.260±0.020、1.225±0.053、1.136±0.004和1.088±0.081;而流式细胞术检测显示,siENST00000414355干扰细胞组72h后HL-60、K562、THP-1和U937的细胞凋亡分别为(21.63±1.130)%、(23.853±2.912)%、(19.13±3.245)%和(22.797±2.165)%,明显高于阴性对照组的(0.041±0.003)%、(0.055±0.002)%、(0.022±0.02)%和(0.01±0.01)%,差异有统计学意义,P<0.05。人骨髓检测结果显示,急性白血病患者骨髓中LncRNA-ENST00000414355的表达为(50.070±16.633),与正常人的(1.300±0.592)比较,明显上调,差异有统计学意义,P<0.01。结论 LncRNA-ENST00000414355在白血病骨髓中高表达,它具有调控AML细胞增殖和细胞凋亡的功能。

OBJECTIVE Increasing evidence suggests that long non-coding RNAs（LncRNAs） are involved in a vari- ety of physiological and pathophysiological processes of cancer, but whether lncRNAs can serve as an important role in a- cute leukemia is still unclear. This study was designed to investigate the effect of I.ncRNA ENST00000414355 in regula ting cell proliferation and apoptosis in acute myeloid leukemia. METHODS HL-60, K562, THP 1 and U937 cells were transfected with si-ENST00000414355 and si-NC,respectively,MTT assay was performed to determine the proliferation, cell apoptosis was assayed by flow cytometry using Annexin V-FITC/PI. Bone marrow（BM） samples were obtained from 10 AML patients and five healthy volunteers in Guangzhou First People＇s Hospital. The LncRNA ENST00000414355 ex- pression of the BM samples was detected using qPCR. RESULTS We found that the expression of LncRNA-ENST- 00000414355 in AML cells was significantly knocked down after 72 h treated with si-ENST00000414355. Down regulation of LncRNA ENST00000414355 in AML cell lines HL-60,K562,THP-1 and U937 inhibited cell proliferation,and induced cell apoptosis. LncRNA-ENST00000414355 expression was markedly up-regulated in AML patients＇ bone marrow sam- ples （50. 070±16. 635） comparing with the healthy volunteers＇ （1. 300±0. 592）（P〈0.01）. CONCLUSIONS Results in dicated that LncRNA-ENST00000414355 level up-regulated in AML and regulates cell proliferation and apoptosis.