MicroRNA Library-Based Functional Screening Identified Androgen-Sensitive miR-216a as a Player in Bicalutamide Resistance in Prostate Cancer

Prostate cancer is a major hormone-dependent tumor affecting men, and is often treated by hormone therapy at the primary stages. Despite its initial efficiency, the disease eventually acquires resistance, resulting in the recurrence of castration-resistant prostate cancer. Recent studies suggest that dysregulation of microRNA (miRNA) function is one of the mechanisms underlying hormone therapy resistance. Identification of critical miRNAs involved in endocrine resistance will therefore be important for developing therapeutic targets for prostate cancer. In the present study, we performed an miRNA library screening to identify anti-androgen bicalutamide resistance-related miRNAs in prostate cancer LNCaP cells. Cells were infected with a lentiviral miRNA library and subsequently maintained in media containing either bicalutamide or vehicle for a month. Microarray analysis determined the amounts of individual miRNA precursors and identified 2 retained miRNAs after one-month bicalutamide treatment. Of these, we further characterized miR-216a, because its function in prostate cancer remains unknown. miR-216a could be induced by dihydrotestosterone in LNCaP cells and ectopic expression of miR-216a inhibited bicalutamide-mediated growth suppression of LNCaP cells. Furthermore, a microarray dataset revealed that the expression levels of miR-216a were significantly higher in clinical prostate cancer than in benign samples. These results suggest that functional screening using an miRNA expression library could be useful for identifying novel miRNAs that contribute to bicalutamide resistance in prostate cancer.

[1]  F. S. French,et al.  Androgen receptor stabilization in recurrent prostate cancer is associated with hypersensitivity to low androgen. , 2001, Cancer research.

[2]  Tao Zhang,et al.  Androgen receptor remains critical for cell-cycle progression in androgen-independent CWR22 prostate cancer cells. , 2006, The American journal of pathology.

[3]  A. Vis,et al.  Key targets of hormonal treatment of prostate cancer. Part 1: the androgen receptor and steroidogenic pathways , 2009, BJU international.

[4]  David J. Arenillas,et al.  JASPAR 2014: an extensively expanded and updated open-access database of transcription factor binding profiles , 2013, Nucleic Acids Res..

[5]  M. Kruhøffer,et al.  miR-345 in Metastatic Colorectal Cancer: A Non-Invasive Biomarker for Clinical Outcome in Non-KRAS Mutant Patients Treated with 3rd Line Cetuximab and Irinotecan , 2014, PloS one.

[6]  K. Horie-Inoue,et al.  MicroRNA-574-3p, identified by microRNA library-based functional screening, modulates tamoxifen response in breast cancer , 2015, Scientific Reports.

[7]  E. Antonarakis,et al.  Novel targeted therapeutics for metastatic castration-resistant prostate cancer. , 2010, Cancer letters.

[8]  C. Croce,et al.  MicroRNA dysregulation in cancer: diagnostics, monitoring and therapeutics. A comprehensive review , 2012, EMBO molecular medicine.

[9]  R. Vessella,et al.  Molecular determinants of resistance to antiandrogen therapy , 2004, Nature Medicine.

[10]  F. S. French,et al.  A mechanism for androgen receptor-mediated prostate cancer recurrence after androgen deprivation therapy. , 2001, Cancer research.

[11]  K. Pienta,et al.  A hierarchical network of transcription factors governs androgen receptor-dependent prostate cancer growth. , 2007, Molecular cell.

[12]  Karen E. Knudsen,et al.  AR, the cell cycle, and prostate cancer , 2008, Nuclear Receptor Signaling.

[13]  H. Frierson,et al.  Activation of Akt Signaling in Prostate Induces a TGFβ Mediated Restraint on Cancer Progression and Metastasis , 2013, Oncogene.

[14]  Chang-Zheng Chen,et al.  MicroRNAs as oncogenes and tumor suppressors. , 2005, The New England journal of medicine.

[15]  A. Tesei,et al.  Prolonged exposure to (R)-bicalutamide generates a LNCaP subclone with alteration of mitochondrial genome , 2014, Molecular and Cellular Endocrinology.

[16]  A. Morin,et al.  SIRNA-Directed In Vivo Silencing of Androgen Receptor Inhibits the Growth of Castration-Resistant Prostate Carcinomas , 2007, PloS one.

[17]  N. Seki,et al.  Novel oncogenic function of mesoderm development candidate 1 and its regulation by MiR-574-3p in bladder cancer cell lines , 2011, International journal of oncology.

[18]  H. Klocker,et al.  Androgen receptor down regulation by small interference RNA induces cell growth inhibition in androgen sensitive as well as in androgen independent prostate cancer cells , 2005, The Journal of Steroid Biochemistry and Molecular Biology.

[19]  D. Feldman,et al.  The development of androgen-independent prostate cancer , 2001, Nature Reviews Cancer.

[20]  D. Tindall,et al.  Mechanisms of androgen-refractory prostate cancer. , 2004, The New England journal of medicine.

[21]  Carsten O. Daub,et al.  pre-miRNA profiles obtained through application of locked nucleic acids and deep sequencing reveals complex 5′/3′ arm variation including concomitant cleavage and polyuridylation patterns , 2011, Nucleic acids research.

[22]  Hsuan-Cheng Huang,et al.  Androgen pathway stimulates MicroRNA‐216a transcription to suppress the tumor suppressor in lung cancer‐1 gene in early hepatocarcinogenesis , 2012, Hepatology.

[23]  D. Bartel MicroRNAs: Target Recognition and Regulatory Functions , 2009, Cell.

[24]  H. Nakagama,et al.  miR‐493 induction during carcinogenesis blocks metastatic settlement of colon cancer cells in liver , 2012, The EMBO journal.

[25]  Andrey Golubov,et al.  Alterations of microRNAs and their targets are associated with acquired resistance of MCF‐7 breast cancer cells to cisplatin , 2010, International journal of cancer.

[26]  R. Dahiya,et al.  Genistein Up-Regulates Tumor Suppressor MicroRNA-574-3p in Prostate Cancer , 2013, PloS one.

[27]  C. Croce Causes and consequences of microRNA dysregulation in cancer , 2009, Nature Reviews Genetics.

[28]  Carlo M. Croce,et al.  Causes and Consequences of MicroRNA Dysregulation , 2012, Cancer journal.

[29]  P. Nelson,et al.  Intraprostatic androgens and androgen-regulated gene expression persist after testosterone suppression: therapeutic implications for castration-resistant prostate cancer. , 2007, Cancer research.

[30]  D. Dykxhoorn MicroRNAs and metastasis: little RNAs go a long way. , 2010, Cancer research.

[31]  Jihan Wang,et al.  Aberrant expression of microRNAs in gastric cancer and biological significance of miR-574-3p. , 2012, International immunopharmacology.

[32]  H. Scher,et al.  Targeting the androgen receptor pathway in prostate cancer. , 2008, Current opinion in pharmacology.

[33]  Judy Lieberman,et al.  Dysregulation of microRNA biogenesis and gene silencing in cancer , 2015, Science Signaling.