环状RNA hsa_circ_0006156在系统性红斑狼疮患者外周血中的表达及其对Raji细胞增殖、凋亡的影响
The significance of circular RNA hsa_circ_0006156 expression in peripheral blood of SLE pa-tients and its effect on proliferation and apoptosis of Raji cells
马庆 1李黎 1李文宇 1侯伟 2林有坤1
作者信息
- 1. 广西医科大学第一附属医院皮肤性病科,南宁 530021
- 2. 中国医学科学院地中海贫血防治研究重点实验室,南宁 530021;广西地中海贫血防治重点实验室,南宁 530021
- 折叠
摘要
目的:研究hsa_circ_0006156对Raji细胞增殖、凋亡的影响及其在系统性红斑狼疮(SLE)发生、发展中的作用.方法:收集广西医科大学第一附属医院皮肤性病科2021年9月至2023年9月收治的55例SLE患者(SLE组),并以40例健康对照者作为对照组(HC组),分离出外周血单个核细胞(PBMC)及CD20+B淋巴细胞,使用实时荧光定量聚合酶链反应(RT-qPCR)法检测hsa_circ_0006156表达水平;采用受试者工作特征曲线(ROC)分析hsa_circ_0006156的诊断效能.以Raji细胞为研究对象,使用慢病毒构建hsa_circ_0006156过表达模型,将细胞分为hsa_circ_0006156过表达组(OE组)、转染空病毒载体的阴性对照组(NC组)以及空白对照组(control组).利用CCK8法检测0 h、6 h、12 h、18 h、24 h、30 h、36 h细胞450 nm吸光度,计算OE组和NC组增殖率.Western blotting实验检测各组Bcl-2、cleaved caspase-3凋亡蛋白表达水平.结果:与HC组比较,SLE组PBMC和CD20+B淋巴细胞中hsa_circ_0006156表达量均明显降低(P＜0.001).ROC曲线分析显示,hsa_circ_0006156区分SLE和HC的ROC曲线下面积(AUC)为0.787 3(95%CI:0.697 5～0.877;P＜0.001),敏感度为97.5%,特异度为50.9%,对SLE诊断Cut-off值为＜0.652.hsa_circ_0006156表达量在无活动或轻度活动与中重度活动患者之间存在显著差异(P=0.0148).CCK8实验结果显示,在36h时,OE组Raji细胞增殖较NC组受到明显抑制(P＜0.001).Western blotting检测结果表明,与NC组和control组比较,OE组Bcl-2蛋白表达水平降低,而cleaved caspase-3蛋白表达水平升高(均P＜0.05).结论:hsa_circ_0006156对SLE具有一定的诊断效能.hsa_circ_0006156可能是通过调节B细胞的增殖、凋亡影响SLE的发生、发展.
Abstract
Objective:To investigate the effect of hsa_circ_0006156 on the proliferation and apoptosis of Raji cells and its role in the occurrence and development of systemic lupus erythematosus(SLE).Methods:A total of 55 SLE patients(SLE group)admitted to the Department of Dermatology and Venereal Diseases,the First Affili-ated Hospital of Guangxi Medical University from September 2021 to September 2023 were collected,and 40 healthy controls were taken as the control group(HC group).Peripheral blood mononuclear cells(PBMC)and CD20+B lymphocytes were isolated.Reverse transcription-quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of hsa_circ_0006156,and receiver operating characteristic(ROC)curve was used to analyze the diagnostic efficacy of hsa_circ_0006156.Taking Raji cells as the research object,the hsa_circ_0006156 overexpression model was constructed by lentivirus.The cells were divided into hsa_circ_0006156 overexpression group(OE group),negative control group(NC group)transfected with empty viral vec-tor,and blank control group(control group).CCK8 assay was used to detect the cell absorbance of 450 nm at 0 h,6 h,12 h,18 h,24 h,30,36 h,and the proliferation rate of OE group and NC group was calculated.Western blot-ting was used to detect the expression levels of Bcl-2 and cleaved caspase-3 apoptotic proteins in each group.Re-sults:Compared with the HC group,the expression of hsa_circ_0006156 in PBMC and CD20+B lymphocytes in the SLE group was significantly decreased(P＜0.001).ROC curve analysis showed that the area under the ROC curve(AUC)of hsa_circ_0006156 distinguishing SLE and HC was 0.7873(95%CI:0.6975-0.877;P＜0.001),sensitivity was 97.5%,specificity was 50.9%,and cut-off value for SLE diagnosis was less than 0.652.There was a significant difference in the expression of hsa_circ_0006156 between patients with no or mild activity and mod-erate to severe activity(P=0.0148).CCK8 assay showed that the proliferation of Raji cells in the OE group was significantly inhibited compared with the NC group at 36 h(P＜0.001).The results of western blotting showed that compared with the NC group and the control group,the expression level of Bcl-2 protein in the OE group was decreased,while the expression level of cleaved caspase-3 protein was increased(all P＜0.05).Conclusion:The hsa_circ_0006156 has certain diagnostic efficacy for SLE.It may affect the occurrence and development of SLE by regulating the proliferation and apoptosis of B cells.
关键词
系统性红斑狼疮/环状RNA/hsa_circ_0006156/Raji细胞Key words
systemic lupus erythematosus/circular RNA/hsa_circ_0006156/Raji cells引用本文复制引用
基金项目
国家自然科学基金(81760561)
国家自然科学基金(82260620)
出版年
2024
国家科技期刊平台
NSTL
万方数据