摘要
目的探讨miR-150-5p调控口腔鳞癌细胞增殖、凋亡、迁移和侵袭的作用及作用机制。方法收集口腔鳞癌组织和癌旁正常组织,体外培养人口腔上皮胶质细胞系HOK和口腔鳞癌细胞系HN6、SCC-25和CAL-27,采用实时荧光定量PCR(quantitative real-time pCR,qRT-PCR)和western blot检测组织样本和细胞中miR-150-5p和胰岛素样生长因子2 mRNA结合蛋白2(insulin-like growth factor 2 mRNA binding protein 2,IGF2BP2)的表达。构建过表达miR-150-5p或沉默表达IGF2BP2的CAL-27细胞,采用Cell Counting Kit-8(CCK8)实验和EdU染色检测细胞增殖活性,流式细胞术检测细胞凋亡率,划痕实验和Transwell实验检测细胞迁移和侵袭能力。双荧光素酶实验验证miR-150-5p与IGF2BP2之间的靶向关系。功能拯救实验分析miR-150-5p/IGF2BP2轴在口腔鳞癌细胞生物学行为中的调控作用。结果口腔鳞癌组织和细胞中miR-150-5p低表达,而IGF2BP2 mRNA和蛋白高表达。与对照组比较,过表达miR-150-5p组和沉默IGF2BP2组细胞的增殖活性均降低,细胞凋亡率升高,细胞划痕愈合率降低,细胞迁移和侵袭数目减少。在CAL-27细胞中miR-150-5p能够与IGF2BP2的3’UTR靶向结合。功能拯救实验显示,与miR-150-5p+pcDNA组比较,miR-150-5p+pcDNA-IGF2BP2组细胞增殖活性增强,细胞凋亡率降低,细胞划痕愈合率升高,细胞迁移和侵袭数目增多。结论miR-150-5p在口腔鳞癌中低表达,过表达miR-150-5p可能通过靶向抑制IGF2BP2基因表达,抑制癌细胞的增殖、迁移和侵袭,促进细胞凋亡。
Objective To investigate the regulatory role of miR-150-5p in the proliferation,apoptosis,migration and invasion of oral squamous cell carcinoma cells,and the underlying mechanism.Methods Oral squamous cell carcinoma tissues and adjacent normal tissues were collected.The human oral epithelial glial cell line HOK and oral squamous cell carcinoma cell lines HN6,SCC-25 and CAL-27 were cultured in vitro.The expression levels of miR-150-5p and insulin-like growth factor-2 mRNA-binding protein 2(IGF2BP2)in tissue samples and cells were detected by quantitative real-time PCR(qRT-PCR)and Western blot.After overexpression of miR_150-5p or knockdown of insulin-like growth factor 2 mRNA binding protein 2(IGF2BP2)in CAL-27 cells,cell proliferation was detected by Cell Counting Kit-8(CCK-8)assay and EdU staining.Cell apoptosis was detected by flow cytometry,and cell migration and invasion were detected by wound healing assay and Transwell assay,respectively.Dual-luciferase reporter gene assay was performed to verify the targeting relationship between miR-150-5p and IGF2BP2.Rescue assay was performed to analyze the regulatory role of the miR-150-5p/IGF2BP2 axis in biological behaviors of oral squamous cell carcinoma cells.Results MiR-150-5p was downregulated in oral squamous cell carcinoma tissues and cells,while IGF2BP2 mRNA was upregulated.Compared with those of the control group,overexpression of miR-150-5p or knockdown of IGF2BP2 significantly decreased the proliferative rate,wound healing rate,and the number of migratory and invasive cells,but significantly increased the apoptotic rate.MiR-150-5p could bind to the 3’UTR of IGF2BP2 in CAL-27 cells.Rescue assay showed that co-overexpression of miR-150-5p and IGF2BP2 significantly increased the proliferative rate,wound healing rate,and the number of migratory and invasive cells,but significantly decreased the apoptotic rate as compared to those overexpressing miR-150-5p.Conclusion MiR-150-5p is downregulated in oral squamous cell carcinoma.Overexpression of miR-150-5p can
作者
田国永
李玲
吕志军
TIAN Guoyong;LI Ling;LV Zhijun(Department of Stomatology,the First Affiliated Hospital of Shandong First Medical University,Shandong,Jinan 271199,China)
出处
《河北医药》
CAS
2023年第11期1621-1627,共7页
Hebei Medical Journal
