Tumor suppressor REIC/Dkk‑3 and its interacting protein SGTA inhibit glucocorticoid receptor to nuclear transport

Iwata,Takehiro; Sadahira,Takuya; Ochiai,Kazuhiko; Ueki,Hideo; Sasaki,Takanori; Haung,Peng; Araki,Motoo; Watanabe,Toyohiko; Nasu,Yasutomo; Watanabe,Masami

doi:10.3892/etm.2020.8819

Tumor suppressor REIC/Dkk‑3 and its interacting protein SGTA inhibit glucocorticoid receptor to nuclear transport

Authors:
- Takehiro Iwata
- Takuya Sadahira
- Kazuhiko Ochiai
- Hideo Ueki
- Takanori Sasaki
- Peng Haung
- Motoo Araki
- Toyohiko Watanabe
- Yasutomo Nasu
- Masami Watanabe
View Affiliations

Affiliations: Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700‑8558, Japan, School of Veterinary Nursing and Technology, Faculty of Veterinary Science, Nippon Veterinary and Life Science University, Tokyo 180‑8602, Collaborative Research Center for Okayama Medical Innovation Center (OMIC), Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700‑8558, Japan
Published online on: May 29, 2020 https://doi.org/10.3892/etm.2020.8819
Pages: 1739-1745

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

REIC/Dkk‑3 is a tumor suppressor, and its expression is significantly downregulated in a variety of human cancer types. A previous study performed yeast two‑hybrid screening and identified the small glutamine‑rich tetratricopeptide repeat‑containing protein α (SGTA), known as a negative modulator of cytoplasmic androgen receptor (AR) signaling, which is a novel interacting partner of REIC/Dkk‑3. The previous study also indicated that the REIC/Dkk‑3 protein interferes with the dimerization of SGTA and then upregulates the AR transport and signaling in human prostate cancer PC3 cells. Since the transport of some steroid receptors to nucleus is conducted similarly by dynein motor‑dependent way, the current study aimed to investigate the role of SGTA and REIC/Dkk‑3 in the transport of other glucocorticoid receptors (GR). In vitro reporter assays for the cytoplasmic GR transport were performed in human prostate cancer PC3 cells and 293T cells. As for the SGTA protein, a suppressive effect on the GR transport to the nucleus was observed in the cells. As for the REIC/Dkk‑3 protein, an inhibitory effect was observed for the GR transport in PC3 cells. Under the depleted condition of SGTA by short‑hairpin (sh)RNA, the downregulation of GR transport by REIC/Dkk‑3 was significantly enhanced compared with the non‑depleted condition in PC3 cells, suggesting a compensatory role of REIC/Dkk‑3 in the SGTA mediated inhibition of GR transport. The current study therefore demonstrated that SGTA inhibited the cytoplasmic transport of GR in 293T and PC3 cells, and REIC/Dkk‑3 also inhibited the cytoplasmic transport of GR in PC3 cells. These results may be used to gain novel insight into the GR transport and signaling in normal and cancer cells.

View Figures

View References

1	Shiota M, Fujimoto N, Kashiwagi E and Eto M: The role of nuclear receptors in prostate cancer. Cells. 8(602)2019.PubMed/NCBI View Article : Google Scholar
2	Shiota M and Eto M: Current status of primary pharmacotherapy and future perspectives toward upfront therapy for metastatic hormone-sensitive prostate cancer. Int J Urol. 23:360–369. 2016.PubMed/NCBI View Article : Google Scholar
3	Kim JY, Yu J, Abdulkadir SA and Chakravarti D: KAT8 regulates androgen signaling in prostate cancer cells. Mol Endocrinol. 30:925–936. 2016.PubMed/NCBI View Article : Google Scholar
4	Thadani-Mulero M, Portella L, Sun S, Sung M, Matov A, Vessella RL, Corey E, Nanus DM, Plymate SR and Giannakakou P: Androgen receptor splice variants determine taxane sensitivity in prostate cancer. Cancer Res. 74:2270–2282. 2014.PubMed/NCBI View Article : Google Scholar
5	Korpal M, Korn JM, Gao X, Rakiec DP, Ruddy DA, Doshi S, Yuan J, Kovats SG, Kim S, Cooke VG, et al: An F876L mutation in androgen receptor confers genetic and phenotypic resistance to MDV3100 (enzalutamide). Cancer Discov. 3:1030–1043. 2013.PubMed/NCBI View Article : Google Scholar
6	Ndibe C, Wang CG and Sonpavde G: Corticosteroids in the management of prostate cancer: A critical review. Curr Treat Options Oncol. 16(6)2015.PubMed/NCBI View Article : Google Scholar
7	Arora VK, Schenkein E, Murali R, Subudhi SK, Wongvipat J, Balbas MD, Shah N, Cai L, Efstathiou E, Logothetis C, et al: Glucocorticoid receptor confers resistance to Antiandrogens by bypassing androgen receptor blockade. Cell. 155:1309–1322. 2013.PubMed/NCBI View Article : Google Scholar
8	Isikbay M, Otto K, Kregel S, Kach J, Cai Y, Vander Griend DJ, Conzen SD and Szmulewiz RZ: Glucocorticoid receptor activity contributes to resistance to androgen-targeted therapy in prostate cancer. Horm Cancer. 5:72–89. 2014.PubMed/NCBI View Article : Google Scholar
9	Xie N, Cheng H, Lin D, Liu L, Yang O, Jia L, Fazli L, Gleave ME, Wang Y, Rennie P and Dong X: The expression of glucocorticoid receptor is negatively regulated by active androgen receptor signaling in prostate tumors. Int J Cancer. 136:E27–E38. 2015.PubMed/NCBI View Article : Google Scholar
10	Lempiäinen JK, Niskanen EA, Vuoti KM, Lampinen RE, Göös H, Varjosalo M and Palvimo JJ: Agonist-specific protein interactomes of glucocorticoid and androgen receptor as revealed by proximity mapping. Mol Cell Proteomics. 16:1462–1474. 2017.PubMed/NCBI View Article : Google Scholar
11	Tsuji T, Miyazaki M, Sakaguchi M, Inoue Y and Namba M: A REIC gene shows down-regulation in human immortalized cells and human tumor-derived cell lines. Biochem Biopsy Res Commum. 268:20–24. 2000.PubMed/NCBI View Article : Google Scholar
12	Hirata T, Watanabe M, Kaku H, Kobayashi Y, Yamada H, Sakaguchi M, Takei K, Huh NH, Nasu Y and Kumon H: REIC/Dkk-3-encoding adenoviral vector as a potentially effective therapeutic agent for bladder cancer. Int J Oncol. 41:559–564. 2012.PubMed/NCBI View Article : Google Scholar
13	Watanabe M, Sakaguchi M, Kinoshita R, Kaku H, Ariyoshi Y, Ueki H, Tanimoto R, Ebara S, Ochiai K, Futami J, et al: A novel gene expression system strongly enhances the anticancer effects of a REIC/Dkk-3-encoding adenoviral vector. Oncol Rep. 31:1089–1095. 2014.PubMed/NCBI View Article : Google Scholar
14	Abarzua F, Sakaguchi M, Tanimoto R, Sonegawa H, Li DW, Edamura K, Kobayashi T, Watanabe M, Kashiwakura T, Kaku H, et al: Heat shock proteins play a crucial role in tumor-specific apoptosis by REIC/Dkk-3. Int J Mol Med. 20:37–43. 2007.PubMed/NCBI
15	Kumon H, Sasaki K, Ariyoshi Y, Sadahira T, Ebara S, Hiraki T, Kanazawa S, Yanai H, Watanabe M and Nasu Y: Ad-REIC gene therapy: Promising results in a patient with metastatic CRPC following chemotherapy. Clin Med Insights Oncol. 9:31–38. 2015.PubMed/NCBI View Article : Google Scholar
16	Abarzua F, Sakaguchi M, Takaishi M, Nasu Y, Kurose K, Ebara S, Miyazaki M, Namba M, Kumon H and Huh NH: Adenovirus-mediated overexpression of REIC/Dkk-3 selectively induces apoptosis in human prostate cancer cells through activation of c-Jun-NH2-kinase. Cancer Res. 65:9617–9622. 2005.PubMed/NCBI View Article : Google Scholar
17	Ochiai K, Morimatsu M, Kato Y, Ishiguro-Oonuma T, Udagawa C, Rungsuriyawiboon O, Azakami D, Michishita M, Ariyoshi Y, Ueki H, et al: Tumor suppressor REIC/Dkk-3 and co-chaperone SGTA: Their interaction and roles in the androgen sensitivity. Oncotaget. 7:3283–3296. 2016.PubMed/NCBI View Article : Google Scholar
18	Liu FH, Wu SJ, Hu SM, Hsiao CD and Wang C: Specific interaction of the 70-kDa heat shock cognate protein with the tetratricopeptide repeats. J Bio Chem. 247:34425–34432. 1999.PubMed/NCBI View Article : Google Scholar
19	Paul A, Garcia YA, Zierer B, Patwardhan C, Gutierrez O, Hildenbrand Z, Harris DC, Balsiger HA, Sivils JC, Johnson JL, et al: The cochaperone SGTA demonstrates regulatory specificity for the androgen, glucocorticoid, and progesterone receptors. J Biol Chem. 289:15297–15308. 2014.PubMed/NCBI View Article : Google Scholar
20	Buchanan G, Ricciardeli C, Harris JM, Prescott J, Yu ZC, Jia L, Butler LM, Marshall VR, Scher HI, Gerald WL, et al: Control of androgen receptor signaling in prostate cancer by the cochaperone small glutamine rich tetraticopeptide repeat containing protein alpha. Cancer Res. 67:10087–10096. 2007.PubMed/NCBI View Article : Google Scholar
21	Philp LK, Butler MS, Hichey TE, Bulter LM, Tilley WD and Day TK: SGTA: A new player in the molecular co-chaperone game. Horm cancer. 4:343–357. 2013.PubMed/NCBI View Article : Google Scholar
22	Kaczmarczyk SJ and Green JE: A single vector containing modified cre recombinase and LOX recombination sequences for inducible tissue-specific amplification of gene expression. Nucleic Acids Res. 29(E56)2001.PubMed/NCBI View Article : Google Scholar
23	Dual-Luciferase^® Reporter Assay System Technical Manual. https://www.promega.com/-/media/files/resources/protocols/technical-manuals/0/dual-luciferase-reporter-assay-system-protocol.pdf.
24	Echeverria PC and Picard D: Molecular chaperones, essential partners of steroid hormone receptors for activity and mobility. Biochim Biophys Acta. 1803:641–649. 2010.PubMed/NCBI View Article : Google Scholar
25	Smith DF and Toft DO: Minireview: The intersection of steroid receptors with molecular chaperones: Observations and questions. Mol Endocrinol. 22:2229–2240. 2008.PubMed/NCBI View Article : Google Scholar
26	Pratt WB, Galigniana MD, Morishima Y and Murphy PJ: Role of molecular chaperones in steroid receptor action. Essays Biochem. 40:41–58. 2004.PubMed/NCBI View Article : Google Scholar
27	Grad I and Picard D: The glucocorticoid responses are shaped by molecular chaperones. Mol Cell Endocrinol. 275:2–12. 2007.PubMed/NCBI View Article : Google Scholar
28	Pérez MH, Cormack J, Mallinson D and Mutungi G: A membrane glucocorticoid receptor mediates the rapid/non-genomic actions of glucocorticoids in mammalian skeletal muscle fibres. J Physiol. 591:5171–5185. 2013.PubMed/NCBI View Article : Google Scholar
29	Cato L, Neeb A, Brown M and Cato AC: Control of steroid receptor dynamics and function by genomic actions of the cochaperones p23 and Bag-1L. Nucl Recept Signal. 12(e005)2014.PubMed/NCBI View Article : Google Scholar
30	Akimirah F, Chen J, Basrawala Z, Xin H and Choubey D: DU-145 and PC-3 human prostate cancer cell lines express androgen receptor: Implications for the androgen receptor functions and regulation. FEBS Lett. 2294–2300. 2006.PubMed/NCBI View Article : Google Scholar