长链非编码RNA人类白细胞抗原复合体18通过微RNA-497-5p/细胞周期蛋白E1轴调节弥漫性大B细胞淋巴瘤的增殖、凋亡和侵袭  

Long noncoding RNA human leukocyte antigen complex 18 regulates proliferation,apoptosis and invasion of diffuse large B-cell lymphoma via the microRNA-497-5p/cyclin E1 axis

作　　者：廖子龙 向国强 陈绘迦 LIAO Zilong;XIANG Guoqiang;CHEN Huijia(Hematological Department,Enshi Tujia and Miao Autonomous Prefecture Central Hospital,Enshi,Hubei 445000,China)

机构地区：[1]恩施土家族苗族自治州中心医院血液病科,湖北恩施445000

出　　处：《安徽医药》2024年第4期794-799,I0006,共7页Anhui Medical and Pharmaceutical Journal

基　　金：恩施土家族苗族自治州中心医院科研项目(CTR20190424)。

摘　　要：目的 探讨长链非编码RNA人类白细胞抗原复合体18(lncRNA HCG18)调控微RNA-497-5p(miR-497-5p)/细胞周期蛋白E1(CCNE1)轴对弥漫性大B细胞淋巴瘤(DLBCL)细胞增殖、凋亡和侵袭的影响。方法 实时荧光定量PCR(qRT-PCR)、蛋白质印迹法分别检测2018年5月至2021年5月收集的恩施土家族苗族自治州中心医院DLBCL病人淋巴组织、良性淋巴结增生病人的淋巴组织、人正常B细胞永生化细胞HMy2.CIR、DLBCL细胞系SU-DHL-1、OCI-LY8、U2932中HCG18、miR-497-5p表达及CCNE1蛋白表达,将OCI-LY8细胞分为Ct组(正常培养的OCI-LY8细胞)、pcDNA组(细胞转染过表达物阴性对照)、pcDNA-HCG18组(细胞转染HCG18过表达物)、si-NC组(细胞转染小干扰RNA阴性对照)、si-HCG18组(细胞转染HCG18小干扰RNA)、si-HCG18+inhibitorNC组(细胞转染HCG18小干扰RNA和抑制物阴性对照)、si-HCG18+miR-497-5p inhibitor组(细胞转染HCG18小干扰RNA和miR-497-5p抑制物),CCK-8法检测细胞增殖,流式细胞术检测细胞凋亡,Transwell检测细胞侵袭,蛋白质印迹法检测CCNE1、增殖细胞核抗原(PCNA)、Bcl-2相关X蛋白(Bax)、基质金属蛋白酶9(MMP-9)蛋白表达,双萤光素酶验证HCG18与miR-497-5p、miR-497-5p与CCNE1的关系。结果 在DLBCL淋巴组织和细胞中,HCG18、CCNE1蛋白高表达,miR-497-5p低表达,且在OCI-LY8细胞中HCG18、CCNE1蛋白表达上调最高,miR-497-5p表达下调最多(P<0.05),因此,以OCI-LY8细胞进行后续研究,与si-NC组比较,si-HCG18组HCG18(0.26±0.03比1.01±0.01)、CCNE1蛋白(0.45±0.03比1.44±0.19)表达降低,miR-497-5p(1.95±0.14比1.03±0.02)表达升高(P<0.05),与pcDNA组比较,pcDNA-HCG18组HCG18(1.96±0.23比1.02±0.01)、CCNE1蛋白(2.33±0.21比1.42±0.18)表达升高,miR-497-5p(0.28±0.02比1.02±0.02)表达降低(P<0.05),与siHCG18组、si-HCG18+inhibitor NC组比较,miR-497-5p表达降低(1.21±0.09比1.95±0.14、1.94±0.13),CCNE1蛋白(0.87±0.08比0.45±0.03、0.44±0.04)表达上调(P<0.05),沉默HCG18可抑制OCI-LY8细Objective To investigate the impacts of long non-coding RNA human leukocyte antigen complex 18(lncRNA HCG18)on the proliferation,apoptosis and invasion of diffuse large B-cell lymphoma(DLBCL)cells by regulating MicroRNA-497-5p(miR-497-5p)/cyclin E1(CCNE1)axis.Methods Real time fluorescence quantitative PCR(qRT PCR)and Western blotting were used to detect the expression of HCG18,miR-497-5p,and CCNE1 protein in lymphoid tissue of DLBCL patients,lymphoid tissue of benign lymph node hyperplasia patients at Enshi Tujia and Miao Autonomous Prefecture Central Hospital,and immortalized human normal B cells HMy2.CIR and DLBCL cell lines SU-DHL-1,OCI-LY8,SU-DHL-1,U2932 were collected from May 2018 to May 2021,respectively;OCI-LY8 cells were assigned into Ct group(normally cultured OCI-LY8 cells),pcDNA group(cells transfected with overexpression neg-ative control),pcDNA-HCG18 group(cells transfected with HCG18 overexpression),si-NC group(cells transfected with small interfer-ing RNA negative control),si-HCG18 group(cells transfected with HCG18 small interfering RNA),si-HCG18+inhibitor NC group(cells transfected with HCG18 small interfering RNA and inhibitor negative control),si-HCG18+miR-497-5p inhibitor group(cells transfected with HCG18 small interfering RNA and miR-497-5p inhibitor).CCK-8 assay was applied to detect cell proliferation;flow cytometry was applied to detect apoptosis;Transwell was applied to detect cell invasion;Western blotting was applied to detect the pro-tein expression of CCNE1,proliferating cell nuclear antigen(PCNA),Bcl-2-associated X protein(Bax),and matrix metalloproteinase 9(MMP-9);double luciferase was used to verify the relationship between HCG18 and miR-497-5p,miR-497-5p and CCNE1.Results In DLBCL lymphoid tissues and cells,HCG18 and CCNE1 proteins were highly expressed and miR-497-5p was lowly expressed,and in OCI-LY8 cells,the upregulation of HCG18 and CCNE1 protein expression was the highest,while the downregulation of miR-497-5p ex-pression was the highest(P<0.05).Therefore,OCI-LY8 cells we

关 键 词：人类白细胞抗原复合体 细胞周期蛋白E 淋巴瘤 大B细胞 弥漫性 增殖细胞核抗原 BCL-2相关X蛋白质 基质金属蛋白酶9 微RNA-497-5p 增殖 侵袭 

分 类 号：R733.1[医药卫生—肿瘤]