hsacircRNA0001478在COPD相关肺动脉高压患者中的表达及其对人肺动脉平滑肌细胞增殖的影响  

Expression of circular RNA hsa_circRNA_0001478 in COPD patients with pulmonary hypertension and its effect on cell proliferation of human pulmonary artery smooth muscle cells

作　　者：莫丽蓉 徐燕 马红红 周玮 董辉[2] 杨朝 陈乾 李萍[1] 耑冰[1] Mo Lirong;Xu Yan;Ma Honghong;Zhou Wei;Dong Hui;Yang Zhao;Chen Qian;Li Ping;Zhuan Bing(Department of Respiratory Medicine,Ningxia Hui Autonomous Region People′s Hospital(Ningxia Hui Autonomous Region People′s Hospital Affiliated to Ningxia Medical University),Yinchuan 750001,China;The Management Center of Scientific Research,General Hospital of Ningxia Medical University,Yinchuan 750001,China;Department of Respiratory and Critical Care Medicine,the Science and Technology Town Hospital of Suzhou,Suzhou 215153,China)

机构地区：[1]宁夏回族自治区人民医院(宁夏医科大学附属自治区人民医院)呼吸内科,银川750001 [2]宁夏医科大学总医院科研管理中心,银川750001 [3]苏州科技城医院呼吸与危重症医学科,苏州215153

出　　处：《国际呼吸杂志》2023年第4期425-433,共9页International Journal of Respiration

基　　金：国家自然科学基金(82160017);江苏省自然科学基金(BK20201183);江苏省"双创人才"(JSSCRC2021568);宁夏自然科学基金(2022AAC03351)。

摘　　要：目的基于基因组高通量测序技术筛选慢性阻塞性肺疾病(COPD)相关肺动脉高压患者血液中异常表达的环状非编码RNAs(circRNAs),并研究显著差异表达的hsacircRNA0001478对人肺动脉平滑肌细胞(hPASMCs)增殖及凋亡的影响。方法本研究为实验研究。采用非随机抽样法选取2021年1月至12月在宁夏回族自治区人民医院呼吸内科住院的COPD肺动脉高压期患者、单纯COPD患者及健康人群的血液标本,采用RNA测序进行测序分析,并运用荧光定量聚合酶链反应(RT-PCR)验证外周血标本中超过10倍差异表达基因。通过RT-PCR验证hPASMCs中hsacircRNA0001478被敲低后的表达水平;运用细胞计数试剂盒实验检测细胞的增殖功能;Hoechst 33342实验通过对细胞核染色确定细胞的状态以检测细胞凋亡;Transwell实验检测细胞侵袭和迁移能力。结果根据测序结果,挑选出在COPD合并肺动脉高压患者外周血中明显高表达的4种circRNAs:hsacircRNA0001478、hsacircRNA0006872、hsacircRNA0001859和hsacircRNA0016070,构建相对应的引物,并通过RT-PCR在COPD合并肺动脉高压患者进行验证,发现仅hsacircRNA0001478表达差异较为明显。构建的si-circ质粒可显著降低hsacircRNA0001478在hPASMCs中的表达,转染si-circ的细胞增殖、迁移及侵袭能力均降低,并促进细胞凋亡。结论hsacircRNA0001478的异常表达对hPASMCs的细胞增殖和细胞凋亡均有影响,这些结果表明hsacircRNA0001478在COPD合并肺动脉高压的发生、发展中可能发挥重要的作用。Objective To screen abnormally-expressed circular non-coding RNA(circRNA)in the blood of chronic obstructive pulmonary disease(COPD)patients based on high-throughput genome sequencing,and to explore the effects of significantly differentially expressed hsa_circRNA_0001478 on human pulmonary artery smooth muscle cells proliferation and apoptosis.Methods This was an experimental study.Using non-random sampling method,the blood samples were collected of COPD patients with pulmonary arterial hypertension and patients with simple COPD hospitalized in the Respiratory Department of the People′s Hospital of Ningxia Hui Autonomous Region from January to December 2021 and the healthy people for check-up during the same period.RNA sequencing(RNA-seq)was used for sequencing analysis,and the real-time fluorescent quantitative polymerase chain reaction(RT-PCR)was used to verify more than 10 times differentially expressed genes in the peripheral blood samples.The expression level of hsa_circRNA_0001478 after knockdown in human pulmonary artery smooth muscle cells(hPASMCs)was verified by RT-PCR;cell counting kit experiment was used to detect cell proliferation;Hoechst 33342 experiment was used to detect cell state and cell apoptosis through nuclear staining;Transwell assay was used to detect cell invasion and migration.Results Based on the sequencing results,four circRNAs that were obviously highly expressed in the peripheral blood of COPD patients with pulmonary arterial hypertension were selected:hsa_circRNA_0001478,hsa_circRNA_0006872,hsa_circRNA_0001859,and hsa_circRNA_0016070,and the corresponding primers were constructed and verified by RT-PCR in COPD patients.It was found that only hsa_circRNA_0001478 showed obvious expression difference.The constructed si-circ plasmid significantly reduced the expression of hsa_circRNA_0001478 in hPASMCs,and cells transfected with si-circ showed reduced proliferation,migration,and invasion ability and increased cell apoptosis.Conclusions The abnormal expression of hsa_circRNA_0001478 a

关 键 词：肺疾病 慢性阻塞性 RNA 环状 肺动脉高压 细胞增殖 细胞凋亡 HsacircRNA0001478 人肺动脉平滑肌细胞 

分 类 号：R544.1[医药卫生—心血管疾病] R563.9[医药卫生—内科学]