Zhou Jianpeng, Gao Feng, Zhang Hua, Xing Mingxuan, Xu Zining, Zhang Ruoyan
Department of Hepatobiliary and Pancreatic Surgery, The First Hospital of Jilin University, Changchun, Jilin, China.
Department of Gastrointestinal Surgery, The First Hospital of Jilin University, Changchun, Jilin, China.
Arch Med Sci. 2019 Nov 11;17(2):481-491. doi: 10.5114/aoms.2019.89650. eCollection 2021.
Previous studies have reported that miR-520b exhibited inhibitory effects on various human tumors, whereas the effects of miR-520b on gallbladder carcinoma (GBC) have remained unclear. To investigate the effects of miR-520b on GBC progression and reveal the underlying mechanisms, this study was performed.
MiR-520b and RAB22A mRNA levels were analyzed by quantitative real-time PCR (qPCR). RAB22A protein level was analyzed via Western blot and immunohistochemical (IHC) analysis. The proliferation, colony formation ability, migration and invasion of NOZ cells were measured via MTT, colony formation, wound healing and transwell invasion assay respectively.
MiR-520b expression level was lower in human GBC tissues than that in neighboring normal tissues. MiR-520b mimic repressed NOZ cell proliferation, colony formation ability, migration and invasion, whereas miR-520b inhibitor exhibited opposite effects. Dual luciferase reporter assay confirmed that miR-520b could bind to the 3'-untranslated regions of RAB22A mRNA. Moreover, RAB22A overexpression significantly abolished the anti-tumor effects of miR-520b in a NOZ cell model. Western blot, qPCR and IHC analysis proved that human GBC tissues showed a higher RAB22A expression level than neighboring normal tissues. Additionally, there was a negative association between miR-520b and RAB22A expression.
MiR-520b had suppressive effects on GBC via targeting RAB22A
先前的研究报道miR-520b对多种人类肿瘤具有抑制作用,而miR-520b对胆囊癌(GBC)的作用仍不清楚。为了研究miR-520b对GBC进展的影响并揭示其潜在机制,进行了本研究。
通过定量实时PCR(qPCR)分析miR-520b和RAB22A mRNA水平。通过蛋白质印迹和免疫组织化学(IHC)分析检测RAB22A蛋白水平。分别通过MTT、集落形成、伤口愈合和Transwell侵袭实验检测NOZ细胞的增殖、集落形成能力、迁移和侵袭能力。
人类GBC组织中miR-520b表达水平低于相邻正常组织。miR-520b模拟物抑制NOZ细胞增殖、集落形成能力、迁移和侵袭,而miR-520b抑制剂则表现出相反的作用。双荧光素酶报告基因实验证实miR-520b可与RAB22A mRNA的3'-非翻译区结合。此外,RAB22A过表达显著消除了miR-520b在NOZ细胞模型中的抗肿瘤作用。蛋白质印迹、qPCR和IHC分析证明,人类GBC组织中RAB22A表达水平高于相邻正常组织。此外,miR-520b与RAB22A表达之间呈负相关。
miR-520b通过靶向RAB22A对GBC具有抑制作用