title: Mir-135a enhances cellular proliferation through post-transcriptionally regulating PHLPP2 and FOXO1 in human bladder cancer.
abstract: BACKGROUND
  Bladder cancer is the most common malignancy in urinary system and the ninth most common malignancy in the world. MicroRNAs (miRNAs) are small, non-coding RNAs that regulate gene expression by targeted repression of transcription and translation and play essential roles during cancer development. We investigated the expression of miR-135a in bladder cancer and explored its bio-function during bladder cancer progression.
  
  
  METHODS
  The expression of miR-135a in bladder cancer cells and tissues are performed by using Real-time PCR assay. Cell viability assay (MTT assay), colony formation assay, anchorage-independent growth ability assay and Bromodeoxyuridine labeling and immunofluorescence (BrdUrd) assay are used to examine cell proliferative capacity and tumorigenicity. Flow cytometry analysis is used to determine cell cycle progression. The expressions of p21, p27, CyclinD1, Ki67, PHLPP2 and FOXO1 are measured by Western blotting assay. Luciferase assay is used to confirm whether FOXO1 is the direct target of miR-135a.
  
  
  RESULTS
  miR-135a is upregulated in bladder cancer cells and tissues. Enforced expression of miR-135a promotes bladder cancer cells proliferation, whereas inhibition of miR-135a reverses the function. Furthermore, for the first time we demonstrated PHLPP2 and FOXO1 are direct targets of miR-135a and transcriptionally down-regulated by miR-135a. Suppression of PHLPP2 or FOXO1 by miR-135a, consisted with dysregulation of p21, p27, Cyclin D1 and Ki67, play important roles in bladder cancer progression.
  
  
  CONCLUSIONS
  Our study demonstrates that miR-135a promotes cell proliferation in bladder cancer by targeting PHLPP2 and FOXO1, and is performed as an onco-miR.
authors.affiliation: Department of Urology, the First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, 510080, PR China. mxpzc1979@163.com.
authors.forename: Xiao Peng
authors.initials: XP
authors.lastname: Mao
authors.affiliation: Oncology Department, PLA458 Hospital, Guangzhou, 510000, China. lawysun@163.com.
authors.forename: Luo Sheng
authors.initials: LS
authors.lastname: Zhang
authors.affiliation: Department of Urology, the First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, 510080, PR China. huangbin2007@yahoo.com.
authors.forename: Bin
authors.initials: B
authors.lastname: Huang
authors.affiliation: Department of Urology, the First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, 510080, PR China. zhoushying@yeah.net.
authors.forename: Shi Ying
authors.initials: SY
authors.lastname: Zhou
authors.affiliation: Department of Urology, the First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, 510080, PR China. junliaogz@163.com.
authors.forename: Jun
authors.initials: J
authors.lastname: Liao
authors.affiliation: Oncology Department, PLA458 Hospital, Guangzhou, 510000, China. lingwuch@126.com.
authors.forename: Ling Wu
authors.initials: LW
authors.lastname: Chen
authors.affiliation: Department of Urology, the First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, 510080, PR China. qiushp@mail.sysu.edu.cn.
authors.forename: Shao Peng
authors.initials: SP
authors.lastname: Qiu
authors.affiliation: Department of Urology, the First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, 510080, PR China. chenjunxindoctor@163.com.
authors.forename: Jun Xing
authors.initials: JX
authors.lastname: Chen
mesh_terms: D001483:Base Sequence; D002453:Cell Cycle / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D019938:Cyclin D1 / Q000378:metabolism; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000378:metabolism; D000071161:Forkhead Box Protein O1; D051858:Forkhead Transcription Factors / Q000235:genetics* / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D019394:Ki-67 Antigen / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008954:Models, Biological; D008969:Molecular Sequence Data; D010749:Phosphoprotein Phosphatases / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic; D001749:Urinary Bladder Neoplasms / Q000235:genetics* / Q000473:pathology*
chemical_list: C530104:CCND1 protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; C085126:FOXO1 protein, human; D000071161:Forkhead Box Protein O1; D051858:Forkhead Transcription Factors; D019394:Ki-67 Antigen; C544965:MIRN135 microRNA, human; D035683:MicroRNAs; D019938:Cyclin D1; D050760:Cyclin-Dependent Kinase Inhibitor p27; C541572:PHLPP2 protein, human; D010749:Phosphoprotein Phosphatases