title: Downregulation of microRNA-182-5p contributes to renal cell carcinoma proliferation via activating the AKT/FOXO3a signaling pathway.
abstract: BACKGROUND
  Emerging evidence has suggested that dysregulation of miR-182-5p may contribute to tumor development and progression in several types of human cancers. However, its role in renal cell carcinoma (RCC) is still unknown.
  
  
  METHODS
  Quantitative RT-PCR was used to quantify miR-182-5p expression in RCC clinical tissues. Bisulfite sequencing PCR was used for DNA methylation analysis. The CCK-8, colony formation, flow cytometry, and a xenograft model were performed. Immunohistochemistry was conducted using the peroxidase and DAB methods. A miR-182-5p target was determined by luciferase reporter assays, quantitative RT-PCR, and Western blotting.
  
  
  RESULTS
  miR-182-5p is frequently down-regulated in human RCC tissues. Epigenetic modulation may be involved in the regulation of miR-182-5p expression. Enforced expression of miR-182-5p in RCC cells significantly inhibited the proliferation and tumorigenicity in vitro and in vivo. Additionally, overexpression of miR-182-5p induced G1-phase arrest via inhibition of AKT/FOXO3a signaling. Moreover, FLOT1 was confirmed as a target of miR-182-5p. Silencing FLOT1 by small interfering RNAs phenocopied the effects of miR-182-5p overexpression, whereas restoration of FLOT1 in miR-182-5p -overexpressed RCC cells partly reversed the suppressive effects of miR-182-5p.
  
  
  CONCLUSIONS
  These findings highlight an important role for miR-182-5p in the pathogenesis of RCC, and restoration of miR-182-5p could be considered as a potential therapeutic strategy for RCC therapy.
authors.forename: Xin
authors.initials: X
authors.lastname: Xu
authors.forename: Jian
authors.initials: J
authors.lastname: Wu
authors.forename: Shiqi
authors.initials: S
authors.lastname: Li
authors.forename: Zhenghui
authors.initials: Z
authors.lastname: Hu
authors.forename: Xianglai
authors.initials: X
authors.lastname: Xu
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhu
authors.forename: Zhen
authors.initials: Z
authors.lastname: Liang
authors.forename: Xiao
authors.initials: X
authors.lastname: Wang
authors.forename: Yiwei
authors.initials: Y
authors.lastname: Lin
authors.forename: Yeqing
authors.initials: Y
authors.lastname: Mao
authors.forename: Hong
authors.initials: H
authors.lastname: Chen
authors.forename: Jindan
authors.initials: J
authors.lastname: Luo
authors.forename: Ben
authors.initials: B
authors.lastname: Liu
authors.affiliation: Department of Urology, First Affiliated Hospital, School of Medicine, Zhejiang University, Qingchun Road 79, Hangzhou 310003, Zhejiang Province, China. zheng_xy@zju.edu.cn.
authors.forename: Xiangyi
authors.initials: X
authors.lastname: Zheng
authors.forename: Liping
authors.initials: L
authors.lastname: Xie
mesh_terms: D000818:Animals; D001483:Base Sequence; D002292:Carcinoma, Renal Cell / Q000235:genetics* / Q000378:metabolism / Q000401:mortality / Q000473:pathology; D049109:Cell Proliferation; D044127:Epigenesis, Genetic; D005260:Female; D000071316:Forkhead Box Protein O3; D051858:Forkhead Transcription Factors / Q000235:genetics* / Q000378:metabolism; D059585:G1 Phase Cell Cycle Checkpoints; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007680:Kidney Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000401:mortality / Q000473:pathology; D008297:Male; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009368:Neoplasm Transplantation; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D016019:Survival Analysis
chemical_list: C499517:FOXO3 protein, human; D000071316:Forkhead Box Protein O3; D051858:Forkhead Transcription Factors; D008565:Membrane Proteins; D035683:MicroRNAs; C538984:Mirn182 microRNA, human; D034741:RNA, Small Interfering; C106354:flotillins; D051057:Proto-Oncogene Proteins c-akt