摘要
目的探究miR-873-5p在结直肠癌(CRC)中表达特征及其与患者预后相关性分析。方法基于TarBase v.7.0数据库筛选靶标mRNA;利用GEO数据库进行差异表达基因筛选,并进行KEGG信号通路富集分析;在结肠癌SW480细胞中过表达miR-873-5p,进行功能实验。miR-873-5p表达水平采用qRT-PCR法测定;细胞增殖检测采用CCK-8法测定;细胞迁移使用Transwell法测定;采用qRT-PCR和Western blotting检测SW-620、LOVO、SW480细胞和正常结肠上皮NCM-460细胞的mRNA和蛋白表达水平;流式细胞仪检测SW480细胞凋亡率;高内涵系统检测SW480细胞线粒体膜电位。结果miR-873-5p在CRC中低表达(P<0.01),而过表达miR-873-5p可使下游分子SF1和NF-κB表达显著增加(P<0.01),并使肿瘤细胞增殖、迁移、凋亡能力显著降低(P<0.01);KEGG通路富集分析显示miR-873-5p下游通路富集于NF-κB通路,预后分析显示,miR-873-5p靶标分子SF1和NF-κB与患者良好呈正相关(P<0.01)。结论miR-873-5p在CRC中低表达,过表达miR-873-5p可抑制结肠癌SW480细胞增殖与迁移,并诱导细胞凋亡,从而可成为CRC潜在的分子靶标。
Objective To explore the expression characteristics of miR-873-5p in colorectal cancer(CRC)and its correlation with patient prognosis.Methods Target mRNA was screened based on TarBase v.7.0 database.Differentially expressed genes were screened using GEO database,and KEGG signaling pathway enrichment analysis was performed.MiR-873-5p was overexpressed in colon cancer SW480 cells for functional testing.The expression level of miR-873-5p was determined by qRT-PCR.Cell proliferation detection was detected by CCK-8 method.Cell migration was measured by Transwell method.qRT-PCR and Western blotting were used to detect mRNA and protein expression levels in SW-620,LOVO,SW480 cells and normal colon epithelial NCM-460 cells.Flow cytometry was used to detect the apoptosis rate of SW480 cells.High-content screening was used to detect mitochondrial membrane potential in SW480 cells.Results MiR-873-5p was underexpressed in CRC(P<0.01),while overexpression of miR-873-5p significantly increased the expression of downstream molecules SF1 and NF-κB(P<0.01),and significantly reduced the proliferation,migration,and apoptosis ability of tumor cells(P<0.01).KEGG pathway enrichment analysis showed that the downstream pathway of miR-873-5p was enriched in NF-κB pathway,and prognostic analysis showed that the target molecules of miR-873-5p,SF1 and NF-κB,were positively correlated with patients(P<0.01).Conclusion MiR-873-5p is underexpressed in CRC,while overexpression of miR-873-5p can inhibit the proliferation and migration of colon cancer SW480 cells,and induce cell apoptosis,thus becoming a potential molecular target for CRC.
作者
赵奕涵
宋亮
胡文婷
邓佳
杨桃
ZHAO Yihan;SONG Liang;HU Wengting;DENG Jia;YANG Tao(The Second Clinical Medical College,Shaanxi University of Chinese Medicine,Xianyang 712046,China;State Key Laboratory of Holistic Integrative Management of Gastrointestinal Cancers,National Clinical Research Center for Digestive Diseases,Xijing Hospital of Digestive Diseases,Air Force Medical University,Xi'an 710032,China;The First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine,Guiyang 550001,China)
出处
《空军军医大学学报》
CAS
2024年第7期775-780,共6页
Journal of Air Force Medical University
基金
国家自然科学基金(82260936)
国家中医药管理局高水平中医药重点学科建设项目(zyyzdxk-2023188)
贵州中医药大学中医脾胃病科技创新人才团队建设项目(贵中医TD[2022]005)
贵州中医药大学博士启动基金(GYZYYFY-BS-2022-01)。
