LncRNA MIAT在颅内动脉瘤患者血清中的表达及靶向调节miR-331-3p对血管平滑肌细胞增殖和凋亡的影响  

Expression of LncRNA MIAT in serum of patients with intracranial aneurysm and the impacts of targeted regulation of miR-331-3p on the proliferation and apoptosis of vascular smooth muscle cells

作　　者：马思伟 董伟[1] 陈思[1] MA Siwei;DONG Wei;CHEN Si(Department of Neurosurgery,Tangshan People's Hospital,Tangshan 063000,Hebei,China)

机构地区：[1]唐山市人民医院神经外科,河北唐山063000

出　　处：《贵州医科大学学报》2024年第7期997-1004,共8页Journal of Guizhou Medical University

基　　金：河北省2024年度医学科学研究课题(20240351);2022年唐山市市级科技计划项目(22130202H)。

摘　　要：目的分析长非编码RNA(LncRNA)心肌梗死相关转录物(MIAT)在颅内动脉瘤(IA)患者血清中的表达,探究其对血管平滑肌细胞(VSMCs)增殖和凋亡的影响及作用机制。方法数字减影血管造影诊断的IA患者15例为IA组,同期无IA家族史的健康参与者15例为对照组(C组),qRT-PCR检测两组受试者血清LncRNA MIAT和miR-331-3p水平;体外培养的VSMCs分为空白对照组(BC组)、过表达LncRNA MIAT阴性对照组(oe-NC组)、过表达LncRNA组(oe-MIAT组)、过表达LncRNA+过表达miR-331-3p阴性对照组(oe-MIAT+miR-331-3p-NC组)及过表达LncRNA+过表达miR-331-3p组(oe-MIAT+miR-331-3p mimic组);采用qRT-PCR检测VSMCs中LncRNA MIAT、miR-331-3p表达,MST测定VSMCs 24、48、72 h时的细胞活力(OD值),EdU染色测定VSMCs增殖,AnnexinⅤ/7-AAD双染法测定VSMCs凋亡,Western blot分析VSMCs中Bax、Bcl-2、cleaved-Caspase-3表达;双荧光素酶报告、RNA免疫共沉淀验证LncRNA MIAT、miR-331-3p靶向关系。结果与C组相比,IA组患者血清LncRNA MIAT相对表达量显著升高,血清miR-331-3p相对表达量显著降低(P<0.05);与BC组及oe-NC组相比,oe-MIAT组、oe-MIAT+miR-331-3p-NC组LncRNA MIAT相对表达量、凋亡率、cleaved-Caspase-3、Bax表达增高,miR-331-3p的相对表达量、24 h、48 h、72 h的OD值、EdU阳性细胞比例、Bcl-2蛋白表达均下降(P<0.05);oe-MIAT+miR-331-3p-mimic组与oe-MIAT+miR-331-3p-NC组、oe-MIAT组相比,凋亡率、cleaved-Caspase-3、Bax表达下降,miR-331-3p的相对表达量、24 h、48 h、72 h的OD值、EdU阳性细胞比例、Bcl-2蛋白水平均显著增高(P<0.05);双荧光素酶报告、RNA免疫共沉淀表明LncRNA MIAT和miR-331-3p之间存在负向调控。结论LncRNA MIAT在IA患者血清中高表达,高表达的LncRNA MIAT可通过下调miR-331-3p表达诱导VSMCs过度凋亡、抑制VSMCs增殖,参与IA发生或进展。Objective To analyze the expression of long non-coding RNA(LncRNA)myocardial infarction-associated transcript(MIAT)in the serum of patients with intracranial aneurysm(IA),and to explore its impact and mechanism on the proliferation and apoptosis of vascular smooth muscle cells(VSMCs).Methods A total of 15 patients with IA diagnosed by digital subtraction angiography were classified as IA group,and 15 healthy participants without family history of IA were enrolled in the control group(C group).The expression levels of serum LncRNA MIAT and miR-331-3p were detected by qRT-PCR in both groups.VSMCs cultured in vitro were divided into blank control group(BC group),overexpressed LncRNA MIAT negative control group(oe-NC group),overexpressed LncRNA group(oe-MIAT group),overexpressed LncRNA+overexpressed miR-331-3p negative control group(oe-MIAT+miR-331-3p-NC group),and overexpressed LncRNA+overexpressed miR-331-3p group(oe-MIAT+miR-331-3p mimic group).The expression of LncRNA MIAT and miR-331-3p in VSMCs was detected by qRT-PCR;the cell viability of VSMCs at 24 h,48 h and 72 h was measured by MST;the proliferation of VSMCs was detected by EdU staining;Annexin V/7-AAD double staining was applied to detect apoptosis of VSMCs;the expression of Bax,Bcl-2 and cleaved-Caspase-3 in VSMCs were analyzed by Western blot;dual-luciferase reporter and RNA co-immunoprecipitation were used to verify the targeting relationship between LncRNA MIAT and miR-331-3p.Results Compared with the C group,the relative expression of LncRNA MIAT in serum of IA group was markedly increased,and the relative expression of miR-331-3p in serum was obviously decreased(P<0.05).Compared with BC group and oe-NC group,the relative expression of LncRNA MIAT,apoptosis rate,and the expression of cleaved-Caspase-3 and Bax in oe-MIAT group and oe-MIAT+miR-331-3p-NC group were increased,while the relative expression of miR-331-3p,the OD value at 24 h,48 h,and 72 h,the proportion of EdU positive cells,and the protein level of Bcl-2 were decreased(P<0.05).Compared wi

关 键 词：颅内动脉瘤 长非编码RNA 心肌梗死相关转录物 血管平滑肌细胞 细胞凋亡 细胞增殖 

分 类 号：R739.41[医药卫生—肿瘤]