miRNA-628-3p对非小细胞肺癌H1299细胞增殖、凋亡及侵袭的影响及其与胰岛素样生长因子1受体的靶向关系  

Effects of miRNA-628-3p on proliferation,apoptosis and invasion of non-small cell lung cancer H1299 cells and its targeting relationship with insulin-like growth factor 1 receptor

作　　者：彭华利[1] 胡同晨 杨帆 张帆[1] 刘威[1] 罗伟 Peng Huali;Hu Tongchen;Yang Fan;Zhang Fan;Liu Wei;Luo Wei(Department of Thoracic and Cardiac Surgery,People's Hospital of Leshan,Leshan 614000,China;Department of Respiratory,People's Hospital of Leshan,Leshan 614000,China)

机构地区：[1]乐山市人民医院胸心外科,乐山614000 [2]乐山市人民医院呼吸科,乐山614000

出　　处：《肿瘤研究与临床》2022年第7期481-486,共6页Cancer Research and Clinic

摘　　要：目的探讨miRNA-628-3p(miR-628-3p)对非小细胞肺癌H1299细胞增殖、凋亡及侵袭的影响及其与胰岛素样生长因子1受体(IGF-1R)的靶向关系。方法设立空白对照组(未经处理的H1299细胞)、miR-NC组(转染空载质粒的H1299细胞)、miR-628-3p-M组(转染含miR-628-3p模拟物序列质粒的H1299细胞)、miR-628-3p-I组(转染含miR-628-3p抑制序列质粒的H1299细胞)。各组细胞培养72 h后,采用四甲基偶氮唑盐(MTT)法检测细胞增殖能力,结晶紫染色测定细胞单克隆形成数,流式细胞术测定细胞凋亡水平,Transwell法测定细胞侵袭能力,反转录、实时荧光定量聚合酶链反应(qRT-PCR)法测定细胞中miR-628-3p、IGF-1R mRNA水平,蛋白质印迹法测定细胞中IGF-1R蛋白水平。结果与空白对照组和miR-NC组比较,miR-628-3p-M组细胞存活率[(42±7)%比(78±6)%、(76±7)%]、单克隆形成数[(235±35)个比(614±89)个、(618±75)个]、侵袭细胞数[(265±85)个比(693±185)个、(703±119)个]、IGF-1R mRNA相对表达量(2.17±0.14比3.38±0.15、3.37±0.13)和IGF-1R蛋白相对表达量(0.34±0.13比0.89±0.19、0.88±0.18)均低(均P<0.05),细胞凋亡率[(9.30±3.51)%比(3.30±1.54)%、(3.10±1.94)%]、miR-628-3p相对表达量(6.93±0.17比3.29±0.15、3.30±0.16)均高(均P<0.05);而miR-628-3p-I组细胞存活率[(90±6)%]、单克隆形成数[(1063±102)个]、侵袭细胞数[(1985±426)个]、IGF-1R mRNA相对表达量(4.30±0.18)和IGF-1R蛋白相对表达量(1.47±0.17)均高(均P<0.05),细胞凋亡率[(0.90±0.20)%]、miR-628-3p相对表达量(1.93±0.18)均低(均P<0.05)。与miR-628-3p-M组比较,miR-628-3p-I组细胞存活率、单克隆形成数、侵袭细胞数、IGF-1R mRNA及蛋白相对表达量均高(均P<0.05),细胞凋亡率、miR-628-3p相对表达量均低(均P<0.05)。结论调节miR-628-3p水平对H1299细胞增殖、迁移、侵袭和凋亡有影响;miR-628-3p与IGF-1R可能有靶向关系。Objective To investigate the effects of miRNA-628-3p(miR-628-3p)on the proliferation,apoptosis and invasion of non-small cell lung cancer H1299 cells and its targeting relationship with insulin-like growth factor 1 receptor(IGF-1R).Methods The blank control group(untreated H1299 cells),miR-NC group(H1299 cells transfected with empty plasmid),miR-628-3p-M group(H1299 cells transfected with miR-628-3p mimic sequence plasmid)and miR-628-3p-I group(H1299 cells transfected with miR-628-3p inhibitory sequence plasmid)were established.The cells in each group were cultured for 72 h,and the cell proliferation ability was detected by methyl thiazol tetrazolium(MTT)method,the number of cell monoclonal formation was determined by crystal violet staining,the level of cell apoptosis was determined by flow cytometry,and the cell invasion ability was determined by Transwell method.The mRNA levels of miR-628-3p and IGF-1R in cells were determined by real-time fluorescence quantitative reverse transcription polymerase chain reaction(qRT-PCR),and the protein level of IGF-1R in cells was determined by Western blotting.Results Compared with the blank control group and miR-NC group,the cell survival rate[(42±7)%vs.(78±6)%,(76±7)%],the number of monoclonal formation[235±35 vs.614±89,618±75],the number of invasive cells[(265±85)cells vs.(693±185)cells,(703±119)cells],relative expression of IGF-1R mRNA(2.17±0.14 vs.3.38±0.15,3.37±0.13)and relative expression of IGF-1R protein(0.34±0.13 vs.0.89±0.19,0.88±0.18)in the miR-628-3p-M group were lower(all P<0.05),but the apoptosis rate[(9.30±3.51)%vs.(3.30±1.54)%,(3.10±1.94)%]and relative expression of miR-628-3p(6.93±0.17 vs.3.29±0.15,3.30±0.16)were higher(all P<0.05);the cell survival rate[(90±6)%],the number of monoclonal formation(1063±102),the number of invasive cells[(1985±426)cells],relative expression of IGF-1R mRNA(4.30±0.18)and relative expression of IGF-1R protein(1.47±0.17)in the miR-628-3p-I group were higher(all P<0.05),but the apoptosis rate[(0.90±0.20)

关 键 词：癌 非小细胞肺 受体 IGF1型 miRNA-628-3p 细胞增殖 细胞凋亡 细胞侵袭 

分 类 号：R734.2[医药卫生—肿瘤]