Liu Shuzhen, An Guoyan, Cao Qing, Li Tong, Jia Xinyu, Lei Lei
Key Laboratory of Resource Biology and Biotechnology in Western China, School of Medicine, Northwest University, Ministry of Education, Xi'an 710069, China.
Health Science Center, Xi'an Jiaotong University, Xi'an 710049, China.
Int J Mol Sci. 2021 May 30;22(11):5877. doi: 10.3390/ijms22115877.
Increasing numbers of miRNAs have been observed as oncogenes or tumor suppressors in colorectal cancer (CRC). It was recently reported that hsa-miR-106b-5p (miR-106b) promoted CRC cell migration and invasion. However, there were also studies showing contradictory results. Therefore, in the present study, we further explore the role of miR-106b and its downstream networks in the carcinogenesis of CRC. We observed that the expression of miR-106b is significantly increased in Pan-Cancer and CRC tissues compared with normal tissues from The Cancer Genome Atlas (TCGA) database. Furthermore, we used Transwell, Cell Counting Kit-8, and colony formation assays to clarify that miR-106b promotes the migratory, invasive, and proliferative abilities of CRC cells. For the first time, we systematically screened the target mRNAs and lncRNAs of miR-106b using TCGA database and the bioinformatics algorithms. Dual-luciferase reporter assay confirmed that NR2F2-AS1 and PLEKHO2 are the direct targets of miR-106b. Furthermore, NR2F2-AS1 acts as a competing endogenous RNA (ceRNA) to regulate PLEKHO2 expression by sponging miR-106b. The results of Gene set enrichment analysis (GSEA) and Western blot indicated that they play important roles in CRC progression by regulating MAPK pathway. Thus, miR-106b/NR2F2-AS1/PLEKHO2/MAPK signaling axis may suggest the potential usage in CRC treatment.
在结直肠癌(CRC)中,越来越多的微小RNA(miRNA)被发现具有致癌基因或肿瘤抑制基因的作用。最近有报道称,人源miR-106b-5p(miR-106b)可促进CRC细胞的迁移和侵袭。然而,也有研究得出了相互矛盾的结果。因此,在本研究中,我们进一步探讨了miR-106b及其下游网络在CRC致癌过程中的作用。我们观察到,与来自癌症基因组图谱(TCGA)数据库的正常组织相比,miR-106b在泛癌组织和CRC组织中的表达显著增加。此外,我们使用Transwell实验、细胞计数试剂盒-8实验和集落形成实验来阐明miR-106b可促进CRC细胞的迁移、侵袭和增殖能力。我们首次使用TCGA数据库和生物信息学算法系统地筛选了miR-106b的靶标mRNA和lncRNA。双荧光素酶报告基因实验证实NR2F2-AS1和PLEKHO2是miR-106b的直接靶标。此外,NR2F2-AS1作为一种竞争性内源RNA(ceRNA),通过吸附miR-106b来调节PLEKHO2的表达。基因集富集分析(GSEA)和蛋白质免疫印迹实验结果表明,它们通过调节丝裂原活化蛋白激酶(MAPK)信号通路在CRC进展中发挥重要作用。因此,miR-106b/NR2F2-AS1/PLEKHO2/MAPK信号轴可能为CRC治疗提供潜在的应用价值。