目的:新的研究强调了microRNAs在癌症发生和各种肿瘤恶性进展中的重要作用。本研究主要探讨miR-377-3p对骨肉瘤(OS)CUL1、上皮-间质转化(EMT)和Wnt/β-catenin通路的生物学效应。
方法:我们进行了定量实时聚合酶链反应(qRT-PCR)以分析OS组织和MG-63细胞中miR-377-3p和CUL1的表达水平。然后,细胞计数试剂盒(CCK)-8和Transwell分析用于检查miR-377-3p在OS细胞生长和转移能力中的功能。同时,荧光素酶报告基因测定用于验证CUL1作为miR-377-3p的直接靶标。然后进行qRT-PCR和Westernblot以检测miR-377-3p对EMT和Wnt/β-catenin通路的影响。建立肿瘤异种移植模型以进一步检查miR-377-3p对体内OS肿瘤发生的影响。
结果:miR-377-3p下调在OS组织和细胞中经常被发现,这与OS患者预后较差有关。功能实验显示miR-377-3p恢复可通过调节EMT和Wnt/β-catenin通路显著抑制OS细胞生长和迁移。此外,荧光素酶报告基因分析显示CUL1作为miR-377-3p的功能靶标。此外,OS组织中CUL1的表达升高也表明OS患者的预后不良。此外,体内miR-377-3p过表达也明显抑制了OS肿瘤的生长。
结论:总的来说,上述所有发现均显示miR-377-3p通过CUL1和EMT以及Wnt/β-catenin通路发挥抗OS功能.这些结果可能有助于临床OS治疗的发展。
OBJECTIVE: Emerging studies highlight the crucial effects of microRNAs on cancer initiation and malignant progression of various tumors. This study focused on the biological effect of miR-377-3p on
CUL1 and epithelial-mesenchymal transition (EMT) and Wnt/β-catenin pathways in osteosarcoma (OS).
METHODS: We performed quantitative real-time polymerase chain reaction (qRT-PCR) to analyze miR-377-3p and
CUL1 expression levels in OS tissues and MG-63 cells. Then, cell counting kit (CCK)-8 and Transwell assay were used to examine the functions of miR-377-3p in OS cell growth and metastasis abilities. Meanwhile, luciferase reporter assay was used to validate
CUL1 as direct target of miR-377-3p. qRT-PCR and Western blot were then carried out to detect the impact of miR-377-3p on EMT and Wnt/β-catenin pathways. Tumor xenograft models were established to further examine the effects of miR-377-3p on OS tumorigenesis in vivo.
RESULTS: miR-377-3p downregulation was frequently identified in OS tissues and cells, which was associated with worse prognosis of OS patients. Functional experiments showed miR-377-3p restoration could dramatically repress OS cell growth and migration by regulation of EMT and Wnt/β-catenin pathways. Moreover, luciferase reporter assay revealed that
CUL1 acted as a functional target of miR-377-3p. Additionally, the elevated
CUL1 expressions in OS tissues also indicated poor prognosis of OS patients. Furthermore, the OS tumor growth was also obviously inhibited by miR-377-3p overexpression in vivo.
CONCLUSIONS: Collectively, all the above findings revealed that miR-377-3p exerted anti-OS functions via CUL1 and EMT and Wnt/β-catenin pathways. These results may contribute to the development of clinical OS treatment.