长链非编码RNA KIAA0125对急性髓系白血病U937细胞增殖和凋亡的影响  

Effects of long non-coding RNA KIAA0125 on proliferation and apoptosis of acute myeloid leukemia U937 cells

作　　者：胡华丽 邓发滑 刘远程 王斯奇 张静馨 禄婷婷 黄海 韦四喜 Hu Huali;Deng Fahua;Liu Yuancheng;Wang Siqi;Zhang Jingxin;Lu Tingting;Huang Hai;Wei Sixi(Clinical Laboratory Center of Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou Province,China;School of Medical Laboratory,Guizhou Medical University,Guiyang 550004,Guizhou Province,China)

机构地区：[1]贵州医科大学附属医院临床检验中心,贵州省贵阳市550004 [2]贵州医科大学医学检验学院,贵州省贵阳市550004

出　　处：《中国组织工程研究》2025年第19期3983-3991,共9页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81960031,82260033,81660027),项目负责人:韦四喜;贵州省科技厅资助项目(20185779-70),项目负责人:韦四喜;贵州医科大学附属医院博士研究启动基金(gyfybsky-2021-29),项目负责人:禄婷婷;贵州医科大学附属医院国家自然科学基金区域基金培育项目(gyfynsfc-2021-48),项目负责人:禄婷婷。

摘　　要：背景:U937细胞可以作为急性髓系白血病细胞模型,用于研究急性髓系白血病的生物学特性、信号通路和治疗靶点。目前虽然已有研究报道长链非编码RNA KIAA0125在急性髓系白血病中呈高表达,但其在U937细胞中的生物学功能尚不清楚,在急性髓系白血病发生发展中的作用机制有待进一步阐明。目的:探讨长链非编码RNA KIAA0125在急性髓系白血病患者外周血中的表达水平及对U937细胞增殖、凋亡的影响。方法:RNA-seq分析急性髓系白血病患者骨髓单核细胞样本,筛选得到差异表达基因——长链非编码RNA KIAA0125,利用qRT-PCR检测长链非编码RNA KIAA0125在急性髓系白血病患者外周血中的表达进行验证,通过GEPIA数据库统计分析173例急性髓系白血病患者和70例健康人骨髓细胞中长链非编码RNA KIAA0125 mRNA的表达与预后的关系。随后使用重组慢病毒技术及CRISPR/Cas9-SAM技术分别构建敲低/过表达长链非编码RNA KIAA0125的U937细胞系,qRT-PCR检测长链非编码RNA KIAA0125敲低/过表达效率。接下来,使用CCK-8实验、流式细胞术及Western blot检测敲低/过表达长链非编码RNA KIAA0125对U937细胞增殖、凋亡的影响。最后,使用Western blot检测敲低/过表达长链非编码RNA KIAA0125对Wnt/β-catenin信号通路相关蛋白的影响。结果与结论:①qRT-PCR结果显示长链非编码RNA KIAA0125在急性髓系白血病患者外周血中呈高表达,GEPIA数据库显示长链非编码RNA KIAA0125在急性髓系白血病患者骨髓细胞中呈高表达,高表达组具有更差的生存期;②敲低组长链非编码RNA KIAA0125的敲低效率为70%,成功构建了稳定敲低长链非编码RNA KIAA0125表达的U937细胞,过表达组长链非编码RNA KIAA0125的表达是Vector组的4倍,成功构建了稳定过表达长链非编码RNA KIAA0125的U937细胞;③敲低长链非编码RNA KIAA0125抑制U937细胞的增殖并促进其凋亡,过表达长链非编码RNA KIAA0125则促BACKGROUND:U937 cells can be used as a cell model for studying the biological characteristics,signaling pathways,and therapeutic targets of acute myeloid leukemia.Although it has been reported that long non-coding RNA KIAA0125 is highly expressed in acute myeloid leukemia,its biological function in U937 cells remains unclear,and its mechanism of action in the occurrence and development of acute myeloid leukemia needs to be further clarified.OBJECTIVE:To investigate the expression level of long non-coding RNA KIAA0125 in peripheral blood of patients with acute myeloid leukemia and its effect on the proliferation and apoptosis of U937 cells.METHODS:RNA-sequencing was used to analyze the bone marrow monocyte samples from acute myeloid leukemia patients,and the differentially expressed gene long non-coding RNA KIAA0125 was screened.The expression of long non-coding RNA KIAA0125 in peripheral blood of patients with acute myeloid leukemia was detected by qRT-PCR.The relationship between long non-coding RNA KIAA0125 mRNA expression and prognosis in bone marrow cells of 173 acute myeloid leukemia patients and 70 healthy people was statistically analyzed by GEPIA database.Subsequently,recombinant lentivirus technology and CRISPR/Cas9-SAM technology were used to construct U937 cell lines with knockdown/overexpression of long non-coding RNA KIAA0125.qRT-PCR was used to detect the knockdown/overexpression efficiency of long non-coding RNA KIAA0125.Next,CCK-8 assay,flow cytometry,and western blot assay were used to detect the effects of knockdown/overexpression of long non-coding RNA KIAA0125 on the proliferation and apoptosis of U937 cells.Finally,western blot assay was used to detect the effect of knockdown/overexpressed long non-coding RNA KIAA0125 on Wnt/β-catenin signaling pathway-related proteins.RESULTS AND CONCLUSION:(1)The results of qRT-PCR showed that long non-coding RNA KIAA0125 was highly expressed in peripheral blood of acute myeloid leukemia patients.The results of GEPIA database showed that long non-coding RN

关 键 词：急性髓系白血病 lncKIAA0125 WNT/Β-CATENIN U937细胞 增殖 

分 类 号：R459.9[医药卫生—治疗学] R34[医药卫生—临床医学] R733.71