Androgen Receptor Blockade Using Enzalutamide Suppresses Long Non-Coding RNA ARLNC1 in Prostate Cancer Cells

Document Type : Short Communication


1 Institute of Health Sciences, Istanbul University, Istanbul, Turkey

2 Department of Basic Oncology, Istanbul University Oncology Institute, Istanbul, Turkey

3 Department of Immunology, Istanbul University Aziz Sancar Institute of Experimental 7 Medicine, Istanbul, Turkey


Prostate cancer (PCa) is a common malignant disease with high mortality rates that develops and progresses in an
androgen-dependent way. In recent years, RNA sequencing enabled identification of many PCa-related long noncoding
RNAs including androgen receptor-regulated long non-coding RNA 1 (ARLNC1) and prostate cancer-associated
transcript 1 (PCAT1). In the present study, our goal was to illuminate expression changes of ARLNC1 and PCAT1 in
the context of androgen stimulation or androgen receptor (AR) blockade with respect to AR expression status. In this
experimental study, LNCaP cells and higher AR-expressing LNCaP-AR++ cells were used as cell models. Cells were
treated with dihydrotestosterone (DHT) as an androgen stimulator and/or enzalutamide as an AR inhibitor. Cell viability
was assessed using annexin V and propidium iodide (PI) staining in flow cytometry. Androgen stimulation prompted
baseline ARLNC1 levels by 53.5-fold in the LNCaP cells (P=0.01) and by 25-fold in the LNCAP-AR+ cells (P=0.18). AR
inhibition by enzalutamide reduced baseline ARLNC1 in LNCaP-AR++ cells by 2-fold (P=0.01), but to a lesser extent
in LNCaP cells. Co-treatment of cells with DHT and enzalutamide led to a remarkable decrease in the DHT effect on
ARLNC1 expression. No specific effect of androgen stimulation or AR blockade on PCAT1 expression was detected.
Our results revealed that the extent of induction of ARLNC1 by androgen is modulated by receptor expression status.
In addition, we determined that AR blockade, via enzalutamide, effectively suppresses ARLNC1 both at baseline and
after induction by DHT.


  1. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2018. CA Cancer J Clin. 2018; 68 (1): 7-30.
  2. Damodaran S, Kyriakopoulos CE, Jarrard DF. Newly diagnosed metastatic prostate cancer: has the paradigm changed? Urol Clin North Am. 2017; 44(4): 611-621.
  3. Ichikawa T, Suzuki H, Ueda T, Komiya A, Imamoto T, Kojima S. Hormone treatment for prostate cancer: current issues and future directions. Cancer Chemother Pharmacol. 2005; 56 Suppl 1: 58-63.
  4. Chandrasekar T, Yang JC, Gao AC, Evans CP. Mechanisms of resistance in castration-resistant prostate cancer (CRPC). Transl Androl Urol. 2015; 4(3): 365-380.
  5. Tran C, Ouk S, Clegg NJ, Chen Y, Watson PA, Arora V, et al. Development of a second generation antiandrogen for treatment of advanced prostate cancer. Science. 2009; 324(5928): 787-790.
  6. Washietl S, Pedersen JS, Korbel JO, Stocsits C, Gruber AR, Hackermüller J, et al. Structured RNAs in the ENCODE selected regions of the human genome. Genome Res. 2007; 17(6): 852-864.
  7. Statello L, Guo CJ, Chen LL, Huarte M. Gene regulation by long noncoding RNAs and its biological functions. Nat Rev Mol Cell Biol. 2021; 22(2): 96-118.
  8. Zhang Y, Pitchiaya S, Cieślik M, Niknafs YS, Tien JC, Hosono Y, et al. Analysis of the androgen receptor-regulated lncRNA landscape identifies a role for ARLNC1 in prostate cancer progression. Nat Genet. 2018; 50(6): 814-824.
  9. Prensner JR, Iyer MK, Balbin OA, Dhanasekaran SM, Cao Q, Brenner JC, et al. Transcriptome sequencing across a prostate cancercohort identifies PCAT-1, an unannotated lincRNA implicated in disease progression. Nat Biotechnol. 2011; 29(8): 742-749.
  10. Prensner JR, Chen W, Han S, Iyer MK, Cao Q, Kothari V, et al. The long non-coding RNA PCAT-1 promotes prostate cancer cell proliferation through cMyc. Neoplasia. 2014; 16(11): 900-908.
  11. Horoszewicz JS, Leong SS, Kawinski E, Karr JP, Rosenthal H, Chu TM, et al. LNCaP model of human prostatic carcinoma. Cancer Res. 1983; 43(4): 1809-1818.
  12. Liu L, Wu L, Gao A, Zhang Q, Lv H, Xu L, et al. The influence of dihydrotestosterone on the development of graves’ disease in female BALB/c mice. Thyroid. 2016; 26(3): 449-457.
  13. Özgür E, Celik AI, Darendeliler E, Gezer U. PCA3 silencing sensitizes prostate cancer cells to enzalutamide-mediated androgen receptor blockade. Anticancer Res. 2017; 37(7): 3631-3637.
  14. Hay CW, Watt K, Hunter I, Lavery DN, MacKenzie A, McEwan IJ. Negative regulation of the androgen receptor gene through a primate- specific androgen response element present in the 5’ UTR. Horm Cancer. 2014; 5(5): 299-311.
  15. Yuan F, Hankey W, Wu D, Wang H, Somarelli J, Armstrong AJ, et al. Molecular determinants for enzalutamide-induced transcription in prostate cancer. Nucleic Acids Res. 2019; 47(19): 10104-10114.
  16. Laiseca JE, Ladelfa MF, Cotignola J, Peche LY, Pascucci FA, Castaño BA, et al. Functional interaction between co-expressed MAGE-A proteins. PLoS One. 2017; 12(5): e0178370.
  17. Hay CW, Watt K, Hunter I, Lavery DN, MacKenzie A, McEwan IJ. Negative regulation of the androgen receptor gene through a primate-specific androgen response element present in the 5’ UTR. Horm Cancer. 2014; 5(5): 299-311.
  18. Arenas-Hernandez M, Vega-Sanchez R. Housekeeping gene expression stability in reproductive tissues after mitogen stimulation. BMC Res Notes. 2013; 6: 285.
  19. Scott LJ. Enzalutamide: a review in castration-resistant prostate cancer affiliations expand. Drugs. 2018; 78(18): 1913-1924.
  20. Liang C, Qi Z, Ge H, Liang C, Zhang Y, Wang Z, et al. Long noncoding RNA PCAT-1 in human cancers: a meta-analysis. Clin Chim Acta. 2018; 480: 47-55.