摘要
目的 探讨LINC_00355通过miR-15a-5p调节PHF19在肺癌侵袭转移中的作用机制。方法 采用A549肺癌细胞作为实验细胞。将细胞分为A549肺癌细胞组、lncRNA-NC组、LINC_00355 mimics组、LINC_00355 inhibitor组,进行不同处理。A549肺癌细胞组:不进行任何处理;lncRNA-NC组:加入空白载体;LINC_00355 mimics组:加入LINC_00355过表达载体;LINC_00355 inhibitor组:加入LINC_00355低表达载体。培养结束后,MTT法测定细胞活力,检测单克隆形成数、迁移和侵袭水平。采用萤光素酶报告基因实验分析LINC_00355对miR-15a-5p的靶向作用;实时荧光定量PCR(qPCR)法检测细胞LINC_00355、miR-15a-5p、PHF19,Western blot检测PHF19蛋白表达水平。结果 与A549肺癌细胞组、lncRNA-NC组比较,LINC_00355 mimics组吸光度(A)值、细胞活力、单克隆形成数、穿膜数、迁移距离升高(P<0.05),凋亡率降低(P<0.05);LINC_00355 inhibitor组A值、细胞活力、单克隆形成数、穿膜数、迁移距离降低,凋亡率升高(P<0.05);与LINC_00355 mimics组比较,LINC_00355 inhibitor组A值、细胞活力、单克隆形成数、穿膜数、迁移距离降低,凋亡率升高(P<0.05)。LINC_00355过表达可明显降低miR-15a-5p-wt的萤光素酶活性(P<0.05);与A549肺癌细胞组、lncRNA-NC组比较,LINC_00355 mimics组细胞LINC_00355、PHF19 mRNA表达上调,miR-15a-5p表达降低(P<0.05),LINC_00355 inhibitor组细胞LINC_00355及PHF19 mRNA和蛋白表达降低,miR-15a-5p表达升高(P<0.05);与LINC_00355 mimics组比较,LINC_00355 inhibitor组细胞LINC_00355 mRNA、PHF19 mRNA和蛋白表达降低,miR-15a-5p表达升高(P<0.05)。结论 LINC_00355过表达促进肺癌细胞的增殖、迁移和侵袭并诱导细胞凋亡,敲低LINC_00355则产生相反的效果;其机制可能与LINC_00355负调控miR-15a-5p进而上调肺癌细胞中PHF19的表达有关。
Objective To investigate the effect of LINC_00355 on lung cancer invasion and metastasis by regulating PHF19 through miR-15a-5p.Methods A549 lung cancer cells were used as experimental cells.The cells were divided into A549 lung cancer cell group,lncRNA-NC group,LINC_00355 mimics group and LINC_00355 inhibitor group,and treated differently.A549 lung cancer cell group,no treatment;lncRNA-NC group,added with blank vector;LINC_00355 mimics group,added with LINC_00355 overexpression vector;LINC_00355 inhibitor group,added with LINC_00355 low expression vector.At the end of the culture,the cell viability was measured by MTT assay,and the number of monoclonal clones formed,migration and invasion levels were detected.Luciferase reporter gene assay was used to analyze the targeting effect of LINC_00355 on miR-15a-5p.Real-time fluorescent quantitative PCR(qPCR)was used to detect LINC_00355,miR-15a-5p,and PHF19,and Western blot was used to detect PHF19 protein expression level.Results Compared with the A549 lung cancer cell group and the lncRNA-NC group,the absorbance(A)value,cell viability,the number of monoclonal clones,the number of transmembrane cells and the migration distance were increased(P<0.05),and the apoptosis rate was decreased(P<0.05)in the LINC_00355 mimics group.In the LINC_00355 inhibitor group,the A value,cell viability,the number of clones formed,the number of transmembrane cells,and the migration distance were decreased,and the apoptosis rate was increased(P<0.05).Compared with the LINC_00355 mimics group,the A value,cell viability,number of clones formed,number of transmembrane cells and migration distance were significantly decreased,and the apoptosis rate was increased in the LINC_00355 inhibitor group(P<0.05).Overexpression of LINC_00355 significantly reduced the luciferase activity of miR-15a-5p-wt(P<0.05).Compared with the A549 lung cancer cell group and the lncRNA-NC group,the expression of LINC_00355 and PHF19 mRNA in the LINC_00355 mimics group was up-regulated,and the expression of miR-15a-5p
作者
徐柯楠
刘静
夏丽
周平
杨海龙
XU Kenan;LIU Jing;XIA Li;ZHOU Ping;YANG Hailong(Department of Thoracic Surgery,Hengshui People′s Hospital,Hengshui,Hebei 053000,China)
出处
《国际检验医学杂志》
CAS
2024年第5期629-634,640,共7页
International Journal of Laboratory Medicine
基金
衡水市市级科技计划项目(2020014054Z)。
