长链非编码RNA LINC01268对急性髓系白血病细胞恶性生物学行为的影响  

Effect of Long Non-Coding RNA LINC01268 on the Malignant Biological Behaviors of Acute Myeloid Leukemia Cells

作　　者：陈婉 干定云 CHEN Wan;GAN Ding-Yun(Department of Hematology&Endocrinology,Wuhan Third Hospital,Wuhan 430060,Hubei Province,China)

机构地区：[1]武汉市第三医院血液内分泌内科,湖北武汉430060

出　　处：《中国实验血液学杂志》2023年第6期1608-1616,共9页Journal of Experimental Hematology

基　　金：湖北省卫生健康委员会科研项目(WJ2021M012);武汉市卫生健康委员会科研项目(WX20B31)。

摘　　要：目的:探讨长链非编码RNA LINC01268对急性髓系白血病(AML)细胞凋亡的影响及其相关机制。方法:采用q RT-PCR检测AML患者外周血样本及AML细胞系HL-60、KG-1中LINC01268和miR-217的表达水平;将HL-60细胞分为pc DNA3.1-NC、pc DNA3.1-LINC01268、si-NC、si-LINC01268、miR-NC、miR-217 mimics、si-LINC01268+inhibitorNC和si-LINC01268+miR-217 inhibitor共8组。采用q RT-PCR检测转染后细胞中LINC01268和miR-217 mRNA的表达;双荧光素酶报告实验检测LINC01268与miR-217的靶向关系;CCK-8法检测细胞活力;流式细胞术检测细胞周期分布和凋亡;Western blot检测细胞周期和凋亡相关蛋白p21、Bcl-2、Bax、caspase-3及PI3K/AKT信号通路相关蛋白表达。结果:LINC01268在AML患者外周血样本、AML细胞系HL-60和KG-1中表达量均明显升高(P<0.05),miR-217表达量明显降低(P<0.05)。与si-NC组和miR-NC组相比,si-LINC01268组和miR-217 mimics组HL-60细胞活力下降(P<0.05)、G1期细胞比例和细胞凋亡率均升高(P<0.05),p21、Bax和caspase-3蛋白表达水平升高(P<0.05),而Bcl-2蛋白表达水平下降(P<0.05)。LINC01268靶向结合并负向调控miR-217的表达,抑制miR-217的表达可部分逆转干扰LINC01268对HL-60细胞活力、细胞周期和凋亡的影响;干扰LINC01268可抑制PI3K/AKT信号通路活性,抑制miR-217的表达能部分逆转干扰LINC01268对PI3K/AKT信号通路的抑制作用。结论:AML细胞中LINC01268呈高表达,miR-217呈低表达。LINC01268可通过靶向miR-217表达,促进PI3K/AKT信号通路活性,升高急性髓系白血病细胞存活率,抑制其凋亡。Objective:To investigate the effect of long non-coding RNA LINC01268 on apoptosis of acute myeloid leukemia(AML)cells and related mechanisms.Methods:The expression levels of LINC01268 and miR-217 in peripheral blood samples from AML patients and AML cell lines HL-60 and KG-1 were detected by qRT-PCR.HL-60 cells were divided into pcDNA3.1-NC,pcDNA3.1-LINC01268,si-NC,si-LINC01268,miR-NC,miR-217 mimics,si-LINC01268+inhibitor-NC and si-LINC01268+miR-217 inhibitor groups.The mRNA expressions of LINC01268 and miR-217 were detected by qRT-PCR.The targeting relationship between LINC01268 and miR-217 was detected by dualluciferase reporter assay.Cell viability was detected by CCK-8 assay.Cell cycle distribution and apoptosis were detected by flow cytometry.The expression of cell cycle and apoptosis-related proteins p21,Bcl-2,Bax,caspase-3 and PI3K/AKT signaling pathway-related proteins were detected by Western blot.Results:The expression of LINC01268 in peripheral blood samples of AML patients and AML cell lines HL-60 and KG-1 was increased(P<0.05),and the expression of miR-217 was decreased(P<0.05).Compared with si-NC group and miR-NC group,the viability of HL-60 cells was decreased in si-LINC01268 group and miR-217 mimics group(P<0.05),the proportion of cells in G1 phase and apoptosis rate were increased(P<0.05),the protein expression levels of p21,Bax and caspase-3 were increased(P<0.05),while the protein expression level of Bcl-2 was decreased(P<0.05).LINC01268 targeted and negatively regulated the expression of miR-217,and inhibiting the expression of miR-217 partially reversed the effects of LINC01268 interference on the viability,cell cycle and apoptosis of HL-60 cells.Interference with LINC01268 could inhibit the activity of PI3K/AKT signaling pathway.Inhibiting the expression of miR-217 could partially reverse the inhibition of LINC01268 interference on PI3K/AKT signaling pathway.Conclusion:LINC01268 is highly expressed and miR-217 is lowly expressed in AML cells.LINC01268 can promote the activity of PI3K/AKT sig

关 键 词：LINC01268 miR-217 急性髓系白血病 PI3K/AKT通路 生物学行为 

分 类 号：R733.7[医药卫生—肿瘤]