miR-7856-5p通过靶向作用3对结直肠癌细胞SW480迁移和增殖的影响

Effect of miR-7856-5p on migration and proliferation of colorectal cancer cell line SW480 by targeting EPHA3

目的探索微小RNA(miRNA,miR)-7856-5p对EPH受体A3(EPHA3)基因表达的调控作用及对结直肠癌细胞SW480迁移和增殖的影响。方法实时定量聚合酶链反应(Real-time PCR)检测结直肠癌组织和细胞系中miR-7856-5p的表达。脂质体转染法分别将miR-7856-5p模拟物和miR-NC转入结直肠癌细胞SW480,分别定义为miR-7856-5p组和miR-NC组。Real-time PCR检测转染效果。Transwell实验和CCK-8实验检测转染后细胞迁移和增殖能力。生物信息学软件预测和双荧光素酶报告基因系统验证miR-7856-5p的靶基因。Real-time PCR和Western blot检测转染后细胞中EPHA3的表达。符合正态分布的计量资料以均数±标准差(Mean±SD)表示,组间比较采用t检验,多组间比较采用单因素方差分析。结果miR-7856-5p在结直肠癌组织的表达明显低于癌旁组织(P<0.01)。miR-7856-5p在结直肠癌细胞系的表达明显低于正常肠黏膜上皮细胞(P<0.05),在SW480细胞中表达最低(P<0.01)。miR-7856-5p组miR-7856-5p的表达明显高于miR-NC组,差异有统计学意义[(9.49±1.09)比(1.06±0.18),P<0.01]。miR-NC组和miR-7856-5p组迁移细胞数量分别为(125.70±14.05)个和(42.01±8.98)个,miR-7856-5p组细胞迁移能力明显下降(P<0.01)。与miR-NC组比较,miR-7856-5p组细胞增殖能力明显降低(P<0.05)。生物信息学软件显示miR-7856-5p的靶基因是EPHA3。双荧光素酶报告基因系统证实miR-7856-5p可靶向结合EPHA3基因(P<0.05)。与miR-NC组比较,miR-7856-5p组SW480细胞中EPHA3的表达明显降低(P<0.05)。结论miR-7856-5p可通过靶向调控EPHA3的表达,抑制结直肠癌细胞SW480的迁移和增殖能力。

Objective To explore the regulation of microRNA(miRNA,miR)-7856-5p on the expression of EPH receptor A3(EPHA3)gene and its effect on the migration and proliferation of colorectal cancer cell line SW480.Methods Real-time quantitative polymerase chain reaction(Real-time PCR)was used to detect the expression of miR-7856-5p in colorectal cancer tissues and cell lines.The miR-7856-5p mimic and miR-NC were transfected into colorectal cancer cell line SW480 by lipofection,respectively,and defined as miR-7856-5p group and miR-NC group,respectively.Real-time PCR was used to detect transfection effects.Transwell assay and CCK-8 assay were used to detect cell migration and proliferation after transfection.The bioinformatics software predicts and the dual luciferase reporter gene system validates the target gene of miR-7856-5p.Real-time PCR and Western blot were used to detect the expression of EPHA3 in the transfected cells.The measurement data in accordamce with normal distribution were expressed as mean±standard deviation,t test was used for inter grap comparison,and single factor analysis of variance was used for multi group comparison.Results The expression of miR-7856-5p in colorectal cancer tissues was significantly lower than that in adjacent tissues(P<0.01).The expression of miR-7856-5p in colorectal cancer cell lines was significantly lower than that in normal intestinal mucosal epithelial cells(P<0.05),and lowest in SW480 cells(P<0.01).The expression of miR-7856-5p in miR-7856-5p group was significantly higher than that in miR-NC group,the difference was statistically significant[(9.49±1.09)vs(1.06±0.18),P<0.01].The number of migrated cells in miR-NC group and miR-7856-5p group were(125.70±14.05)and(42.01±8.98),respectively,and the migration ability of miR-7856-5p group was significantly decreased(P<0.01).Compared with the miR-NC group,the cell proliferation ability of the miR-7856-5p group was significantly decreased(P<0.05).Bioinformatics software showed that the target gene for miR-7856-5p was EPHA3.The dual luciferase reporter gene system confirmed that miR-7856-5p can target the EPHA3 gene(P<0.05).Compared with the miR-NC group,the expression of EPHA3 in the SW480 cells of the miR-7856-5p group was significantly decreased(P<0.05).Conclusion miR-7856-5p can inhibit the migration and proliferation of colorectal cancer cell SW480 by regulating the expression of EPHA3.