| 廖子龙,向国强,陈绘迦.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[J].安徽医药,2024,28(4):794-799. |
| 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 |
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| DOI：10.3969/j.issn.1009-6469.2024.04.034 |
| 中文关键词: 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、M，则呈， |
| 英文关键词: Human leukocyte antigen complex group Cyclin E Lymphoma,large B-cell,diffuse Proliferating cell nuclear anti? gen Bcl-2-associated X protein Matrix metalloproteinase 9 MicroRNA-497-5p Proliferation, invasion |
| 基金项目:恩施土家族苗族自治州中心医院科研项目( CTR20190424) |
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| 中文摘要: |
| 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)、 pcD? NA-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-497-5，p(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细胞增殖、侵袭行为及 PCNAMP-9蛋白表达，诱导细胞凋亡及 Bax蛋白表达，而上调 HCG18相反趋势，下调 miR-497-5p逆转了沉默 HCG18对 OCI-LY8细胞增殖、侵袭、凋亡的影响， HCG18靶向调控 miR-497-5p/CCNE1。结论沉默 HCG18可能通过调控 miR-497-5p/CCNE1抑制 OCI-LY8细胞增殖、侵袭，诱导细胞凋亡。 |
| 英文摘要: |
| 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-4975p)/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 lymphnode hyperplasia patients at Enshi Tujia and Miao Autonomous Prefecture Central Hospital, and immortalized human normal B cellsHMy2. 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; flowcytometry 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 were used for subsequent research; compared with the si-NC group, the ex? pression of HCG18 [(0.26±0.03) vs. (1.01±0.01)] and CCNE1 protein [(0.45±0.03) vs.(1.44±0.19)] decreased, and the expression of miR497-5p [(1.95±0.14) vs. (1.03±0.02)] increased in si-HCG18 group (P<0.05); compared with pcDNA group, the expression of HCG18 [(1.96±0.23) vs. (1.02±0.01)] and CCNE1 protein [(2.33±0.21) vs. (1.42±0.18)] increased, and the expression of miR-497-5p [(0.28± 0.02) vs. (1.02±0.02)] decreased in the pcDNA-HCG18 group (P<0.05); compared with si-HCG18 group and si-HCG18+inhibitor NC group, the change of HCG18 expression was not significantly different (P>0.05), and the expression of miR-497-5p decreased [(1.21 ± 0.09) vs. (1.95±0.14), (1.94±0.13)], CCNE1 protein [(0.87±0.08) vs. (0.45±0.03), (0.44±0.04)] increased in si-HCG18+miR-497-5p in? hibitor group (P<0.05); down-regulation of HCG18 could inhibit OCI-LY8 cell proliferation, invasion, PCNA, MMP-9 protein expres? sions , and promote cell apoptosis and Bax protein expression, while up-regulation of HCG18 showed the opposite trend; down-regula? tion of miR-497-5p reversed the effects of silencing HCG18 on the proliferation, invasion and apoptosis of OCI-LY8 cells. HCG18 tar? geted the miR-497-5p/CCNE1.Conclusion Silencing HCG18 may inhibit the proliferation and invasion and induce cell apoptosis of OCI-LY8 cells by regulating miR-497-5p/CCNE1. |
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