MicroRNA-185 downregulates androgen receptor expression in the LNCaP prostate carcinoma cell line

Liu,Chunyan; Chen,Zhaobo; Hu,Xiaoyan; Wang,Lina; Li,Chaoyang; Xue,Jing; Zhang,Pengju; Chen,Weiwen; Jiang,Anli

doi:10.3892/mmr.2015.3332

MicroRNA-185 downregulates androgen receptor expression in the LNCaP prostate carcinoma cell line

Authors:
- Chunyan Liu
- Zhaobo Chen
- Xiaoyan Hu
- Lina Wang
- Chaoyang Li
- Jing Xue
- Pengju Zhang
- Weiwen Chen
- Anli Jiang
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Affiliations: Department of Biochemistry and Molecular Biology, Medical School of Shandong University, Jinan, Shandong 250012, P.R. China
Published online on: February 11, 2015 https://doi.org/10.3892/mmr.2015.3332
Pages: 4625-4632

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Abstract

The present study aimed to investigate whether microRNA (miR)‑185 downregulated androgen receptor expression in the LNCaP prostate carcinoma cell line. Human prostate cancer (PCa) LNCaP cells were cultured and transfected with synthetic has‑miR‑185 mimic or inhibitor. The transfected cells were subsequently evaluated with a viability assay, nuclear staining, reverse transcription quantitative polymerase chain reaction (RT‑qPCR), dual luciferase assay and western blot analysis. The results of the western blot analysis and RT‑qPCR indicated that transfection with an miR‑185 mimic markedly reduced the androgen receptor (AR) protein expression levels in LNCaP cells, whereas transfection with an miR‑185 inhibitor increased the protein expression of AR in the LNCaP cells. The results of the luciferase reporter assay demonstrated that the predicted target site in the AR 3' untranslated regions was a specific functional binding site for miR‑185, and that AR was a direct target of miR‑185. In addition, downregulation of AR by miR‑185 impaired the interaction between AR and androgen response element, and downregulated the expression of the AR target gene prostate specific antigen. Data also suggested that the downregulation of AR mediated by miR‑185, inhibited the proliferation and induced the apoptosis of the LNCaP cells. Therefore, the results of the present study suggested that miR‑185 may be a potential negative modulator of AR‑mediated signaling and may act as a tumor suppressor in prostate cancer cells.

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1	Jemal A, Siegel R, Xu J and Ward E: Cancer statistics, 2010. CA Cancer J Clin. 60:277–300. 2010. View Article : Google Scholar : PubMed/NCBI
2	Jemal A, Siegel R, Ward E, Hao Y, Xu J and Thun MJ: Cancer statistics, 2009. CA Cancer J Clin. 59:225–249. 2009. View Article : Google Scholar : PubMed/NCBI
3	Dehm SM and Tindall DJ: Androgen receptor structural and functional elements: role and regulation in prostate cancer. Mol Endocrinol. 21:2855–2863. 2007. View Article : Google Scholar : PubMed/NCBI
4	Buchanan G, Irvine RA, Coetzee GA and Tilley WD: Contribution of the androgen receptor to prostate cancer predisposition and progression. Cancer Metastasis Rev. 20:207–223. 2001. View Article : Google Scholar
5	Agoulnik IU and Weigel NL: Androgen receptor action in hormone-dependent and recurrent prostate cancer. J Cell Biochem. 99:362–372. 2006. View Article : Google Scholar : PubMed/NCBI
6	Taplin ME and Balk SP: Androgen receptor: a key molecule in the progression of prostate cancer to hormone independence. J Cell Biochem. 91:483–490. 2004. View Article : Google Scholar : PubMed/NCBI
7	Feldman BJ and Feldman D: The development of androgen-independent prostate cancer. Nat Rev Cancer. 1:34–45. 2001. View Article : Google Scholar
8	Pienta KJ and Bradley D: Mechanisms underlying the development of androgen-independent prostate cancer. Clin Cancer Res. 12:1665–1671. 2006. View Article : Google Scholar : PubMed/NCBI
9	Chen Y, Sawyers CL and Scher HI: Targeting the androgen receptor pathway in prostate cancer. Curr Opin Pharmacol. 8:440–448. 2008. View Article : Google Scholar : PubMed/NCBI
10	Knudsen KE and Scher HI: Starving the addiction: new opportunities for durable suppression of AR signaling in prostate cancer. Clin Cancer Res. 15:4792–4798. 2009. View Article : Google Scholar : PubMed/NCBI
11	Eder IE, Culig Z, Ramoner R, Thurnher M, Putz T, Nessler-Menardi C, Tiefenthaler M, Bartsch G and Klocker H: Inhibition of LncaP prostate cancer cells by means of androgen receptor antisense oligonucleotides. Cancer Gene Ther. 7:997–1007. 2000. View Article : Google Scholar : PubMed/NCBI
12	Zegarra-Moro OL, Schmidt LJ, Huang H and Tindall DJ: Disruption of androgen receptor function inhibits proliferation of androgen-refractory prostate cancer cells. Cancer Res. 62:1008–1013. 2002.PubMed/NCBI
13	Wright ME, Tsai MJ and Aebersold R: Androgen receptor represses the neuroendocrine transdifferentiation process in prostate cancer cells. Mol Endocrinol. 17:1726–1737. 2003. View Article : Google Scholar : PubMed/NCBI
14	Chen CD, Welsbie DS, Tran C, et al: Molecular determinants of resistance to antiandrogen therapy. Nat Med. 10:33–39. 2004. View Article : Google Scholar : PubMed/NCBI
15	Hååg P, Bektic J, Bartsch G, Klocker H and Eder IE: Androgen receptor down regulation by small interference RNA induces cell growth inhibition in androgen sensitive as well as in androgen independent prostate cancer cells. J Steroid Biochem Mol Biol. 96:251–258. 2005. View Article : Google Scholar : PubMed/NCBI
16	Compagno D, Merle C, Morin A, Gilbert C, Mathieu JR, Bozec A, Mauduit C, Benahmed M and Cabon F: SIRNA-directed in vivo silencing of androgen receptor inhibits the growth of castration-resistant prostate carcinomas. PLoS One. 2:e10062007. View Article : Google Scholar : PubMed/NCBI
17	Di Giacomo G, Koss M, Capellini TD, Brendolan A, Pöpperl H and Selleri L: Spatio-temporal expression of Pbx3 during mouse organogenesis. Gene Expr Patterns. 6:747–757. 2006. View Article : Google Scholar : PubMed/NCBI
18	Lichtenauer UD, Duchniewicz M, Kolanczyk M, et al: Pre-B-cell transcription factor 1 and steroidogenic factor 1 synergistically regulate adrenocortical growth and steroidogenesis. Endocrinology. 148:693–704. 2007. View Article : Google Scholar
19	Carleton M, Cleary MA and Linsley PS: MicroRNAs and cell cycle regulation. Cell Cycle. 6:2127–2132. 2007. View Article : Google Scholar : PubMed/NCBI
20	Schmittgen TD: Regulation of microRNA processing in development, differentiation and cancer. J Cell Mol Med. 12:1811–1819. 2008. View Article : Google Scholar : PubMed/NCBI
21	Bartel DP: MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
22	Ambros V: MicroRNA pathways in flies and worms: growth, death, fat, stress and timing. Cell. 113:673–676. 2003. View Article : Google Scholar : PubMed/NCBI
23	Hwang HW and Mendell JT: MicroRNAs in cell proliferation, cell death, and tumorigenesis. Br J Cancer. 96(Suppl): R40–R44. 2007.PubMed/NCBI
24	Wiemer EA: The role of microRNAs in cancer: no small matter. Eur J Cancer. 43:1529–1544. 2007. View Article : Google Scholar : PubMed/NCBI
25	Takahashi Y, Forrest AR, Maeno E, Hashimoto T, Daub CO and Yasuda J: Mir-107 and mir-185 can induce cell cycle arrest in human non small cell lung cancer cell lines. PLoS One. 4:e66772009. View Article : Google Scholar : PubMed/NCBI
26	Imam JS, Buddavarapu K, Lee-Chang JS, Ganapathy S, Camosy C, Chen Y and Rao MK: MicroRNA-185 suppresses tumor growth and progression by targeting the six1 oncogene in human cancers. Oncogene. 29:4971–4979. 2010. View Article : Google Scholar : PubMed/NCBI
27	Liu M, Lang N, Chen X, et al: MiR-185 targets RhoA and Cdc42 expression and inhibits the proliferation potential of human colorectal cells. Cancer Lett. 301:151–160. 2011. View Article : Google Scholar
28	Akçakaya P, Ekelund S, Kolosenko I, et al: MiR-185 and miR-133b deregulation is associated with overall survival and metastasis in colorectal cancer. Int J Oncol. 39:311–318. 2011.PubMed/NCBI
29	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
30	Akao Y, Nakagawa Y and Naoe T: let-7 microRNA functions as a potential growth suppressor in human colon cancer cells. Biol Pharm Bull. 9:903–906. 2006. View Article : Google Scholar
31	Majid S, Dar AA, Saini S, et al: miR-23b represses proto-oncogene Src kinase and functions as methylation-silenced tumor suppressor with diagnostic and prognostic significance in prostate cancer. Cancer Res. 72:6435–6446. 2012. View Article : Google Scholar : PubMed/NCBI
32	Ni Y, Meng L, Wang L, Dong W, Shen H, Wang G, Liu Q and Du J: MicroRNA-143 functions as a tumor suppressor in human esophageal squamous cell carcinoma. Gene. 517:197–204. 2013. View Article : Google Scholar : PubMed/NCBI
33	Wu N, Lin X, Zhao X, Zheng L, Xiao L, Liu J, Ge L and Cao S: MiR-125b acts as an oncogene in glioblastoma cells and inhibits cell apoptosis through p53 and p38MAPK-independent pathways. Br J Cancer. 109:2853–2863. 2013. View Article : Google Scholar : PubMed/NCBI
34	Zhang B, Pan X, Cobb GP and Anderson TA: MicroRNAs as oncogenes and tumor suppressors. Dev Biol. 302:1–12. 2007. View Article : Google Scholar
35	Liu H, Brannon AR, Reddy AR, et al: Identifying mRNA targets of microRNA dysregulated in cancer: with application to clear cell renal cell carcinoma. BMC Syst Biol. 4:512010. View Article : Google Scholar : PubMed/NCBI
36	Abaan OD and Toretsky JA: PTPl1: a large phosphatase with a split personality. Cancer Metastasis Rev. 27:205–214. 2008. View Article : Google Scholar : PubMed/NCBI
37	Mangelsdorf DJ, Thummel C, Beato M, et al: The nuclear receptor superfamily: the second decade. Cell. 83:835–839. 1995. View Article : Google Scholar : PubMed/NCBI
38	Heinlein CA and Chang C: Androgen receptor (AR) coregulators: an overview. Endocr Rev. 23:175–200. 2002. View Article : Google Scholar : PubMed/NCBI
39	Chmelar R, Buchanan G, Need EF, Tilley W and Greenberg NM: Androgen receptor coregulators and their involvement in the development and progression of prostate cancer. Int J Cancer. 120:719–733. 2007. View Article : Google Scholar
40	Chen S, Song CS, Lavrovsky Y, Bi B, Vellanoweth R, Chatterjee B and Roy AK: Catalytic cleavage of the androgen receptor messenger RNA and functional inhibition of androgen receptor activity by a hammerhead ribozyme. Mol Endocrinol. 12:1558–1566. 1998. View Article : Google Scholar : PubMed/NCBI
41	Eder IE, Hoffmann J, Rogatsch H, Schafer G, Zopf D, Bartsch G and Klocker H: Inhibition of LNCaP prostate tumor growth in vivo by an antisense oligonucleotide directed against the human androgen receptor. Cancer Gene Ther. 9:117–125. 2002. View Article : Google Scholar : PubMed/NCBI
42	Cheng H, Snoek R, Ghaidi F, Cox ME and Rennie PS: Short hairpin RNA knockdown of the androgen receptor attenuates ligand independent activation and delays tumor progression. Cancer Res. 66:10613–10620. 2006. View Article : Google Scholar : PubMed/NCBI
43	Liao X, Tang S, Thrasher JB, Griebling TL and Li B: Small-interfering RNA-induced androgen receptor silencing leads to apoptotic cell death in prostate cancer. Mol Cancer Ther. 4:505–515. 2005. View Article : Google Scholar : PubMed/NCBI
44	Lim AC and Attard G: Improved therapeutic targeting of the androgen receptor: rational drug design improves survival in castration-resistant prostate cancer. Curr Drug Targets. 14:408–419. 2013. View Article : Google Scholar : PubMed/NCBI