Silencing of <em>ERR&alpha;</em> gene represses cell proliferation and induces apoptosis in human skin fibroblasts

Nanashima,Naoki; Norikura,Toshio; Nakano,Manabu; Hata,Chie; Horie,Kayo

doi:10.3892/mmr.2024.13370

Silencing of ERRα gene represses cell proliferation and induces apoptosis in human skin fibroblasts

Authors:
- Naoki Nanashima
- Toshio Norikura
- Manabu Nakano
- Chie Hata
- Kayo Horie
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Affiliations: Department of Nutrition, Faculty of Health Science, Aomori University of Health and Welfare, Aomori 030‑8505, Japan, Department of Bioscience and Laboratory Medicine, Hirosaki University Graduate School of Health Sciences, Hirosaki, Aomori 036‑8564, Japan, Department of Biomedical Data Intelligence, Graduate School of Medicine, Kyoto University, Kyoto 606‑8397, Japan
Published online on: October 18, 2024 https://doi.org/10.3892/mmr.2024.13370
Article Number: 6
Copyright: © Nanashima et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Estrogen‑related receptor (ERR) is an orphan nuclear receptor structurally akin to the estrogen receptor. ERR is expressed in tissues with active energy metabolism and regulates intracellular metabolic functions. Additionally, ERRs are known to be strongly expressed in the epidermis of skin tissue, but their functions are unknown. The present study investigated the function of ERRα in human skin fibroblasts. ERRα expressed in human dermal fibroblast TIG113 was knocked down using small interfering (si)RNA and gene expression was comprehensively analyzed using microarrays 48 h later. Pathway analysis was performed using Wikipathways on genes exhibiting expression changes of ≥1.5‑fold. Expression of cell cycle‑related and apoptosis‑related genes was compared using reverse transcription‑quantitative PCR. After treating TIG113 cells with siERRα for 72 h, cell proliferation was assessed using the Cell Counting Kit‑8 or a scratch wound healing assay and apoptotic cells were measured using the Poly Caspase Assay Kit. Cell cycle analysis was performed using flow cytometry. The expression of the ERRα gene was suppressed by siRNA. The expression of genes associated with cell cycle‑related pathways were decreased while that of those associated with apoptosis‑related pathways increased. Furthermore, the expression of cell cycle‑related genes such as cell division cycle 25C, cyclin E and cyclin B1 was decreased and the expression of apoptosis‑related genes such as caspase3 and Fas cell surface death receptor was increased. Cell proliferation was suppressed and the number of apoptotic cells increased ~2‑fold in ERRα‑knockdown TIG113 cells. Cell cycle analysis revealed that the number of cells in the Sub‑G₁phase increased and that in the S and G₂/M phases decreased. The present study suggested that ERRα is an essential for the survival of human skin fibroblasts.

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1	Giguère V, Yang N, Segui P and Evans RM: Identification of a new class of steroid hormone receptors. Nature. 331:91–94. 1988. View Article : Google Scholar
2	Horard B and Vanacker JM: Estrogen receptor-related receptors: Orphan receptors desperately seeking a ligand. J Mol Endocrinol. 31:349–357. 2003. View Article : Google Scholar
3	Huss JM, Imahashi K, Dufour CR, Weinheimer CJ, Courtois M, Kovacs A, Giguère V, Murphy E and Kelly DP: The nuclear receptor ERRalpha is required for the bioenergetic and functional adaptation to cardiac pressure overload. Cell Metab. 6:25–37. 2007. View Article : Google Scholar
4	Deblois G and Giguère V: Functional and physiological genomics of estrogen-related receptors (ERRs) in health and disease. Biochim Biophys Acta. 1812:1032–1040. 2011. View Article : Google Scholar
5	Fan W, He N, Lin CS, Wei Z, Hah N, Waizenegger W, He MX, Liddle C, Yu RT, Atkins AR, et al: ERRγ promotes angiogenesis, mitochondrial biogenesis, and oxidative remodeling in PGC1α/β-deficient muscle. Cell Rep. 22:2521–2529. 2018. View Article : Google Scholar
6	Schreiber SN, Emter R, Hock MB, Knutti D, Cardenas J, Podvinec M, Oakeley EJ and Kralli A: The estrogen-related receptor alpha (ERRalpha) functions in PPARgamma coactivator 1alpha (PGC-1alpha)-induced mitochondrial biogenesis. Proc Natl Acad Sci USA. 101:6472–6477. 2004. View Article : Google Scholar
7	Vernier M and Giguère V: Aging, senescence and mitochondria: The PGC-1/ERR axis. J Mol Endocrinol. 66:R1–R14. 2021. View Article : Google Scholar
8	Luo J, Sladek R, Bader JA, Matthyssen A, Rossant J and Giguère V: Placental abnormalities in mouse embryos lacking the orphan nuclear receptor ERR-beta. Nature. 388:778–782. 1997. View Article : Google Scholar
9	Giguère V: To ERR in the estrogen pathway. Trends Endocrinol Metab. 13:220–225. 2002. View Article : Google Scholar
10	Brincat MP, Baron YM and Galea R: Estrogens and the skin. Climacteric. 8:110–123. 2005. View Article : Google Scholar
11	Stern R and Maibach HI: Hyaluronan in skin: Aspects of aging and its pharmacologic modulation. Clin Dermatol. 26:106–122. 2008. View Article : Google Scholar
12	Thornton MJ: Estrogens and aging skin. Dermatoendocrinol. 5:264–270. 2013. View Article : Google Scholar
13	Haczynski J, Tarkowski R, Jarzabek K, Slomczynska M, Wolczynski S, Magoffin DA, Jakowicki JA and Jakimiuk AK: Human cultured skin fibroblasts express estrogen receptor alpha and beta. Int J Mol Med. 10:149–153. 2002.
14	Saito K and Cui H: Emerging roles of estrogen-related receptors in the brain: Potential interactions with estrogen signaling. Int J Mol Sci. 19:10912018. View Article : Google Scholar
15	Bertil E, Bolzinger MA, André V, Rousselle P and Damour O: Expression of oestrogen-related receptor alpha in human epidermis. Exp Dermatol. 17:208–213. 2008. View Article : Google Scholar
16	Krahn-Bertil E, Dos Santos M, Damour O, Andre V and Bolzinger MA: Expression of estrogen-related receptor beta (ERRbeta) in human skin. Eur J Dermatol. 20:719–723. 2010.
17	Krahn-Bertil E, Bolzinger MA, Andre V, Orly I, Kanitakis J, Rousselle P and Damour O: Expression of estrogen-related receptor gamma (ERRgamma) in human skin. Eur J Dermatol. 18:427–432. 2008.
18	Liu D, Zhang Z, Gladwell W and Teng CT: Estrogen stimulates estrogen-related receptor alpha gene expression through conserved hormone response elements. Endocrinology. 144:4894–4904. 2003. View Article : Google Scholar
19	Fujimoto J, Nakagawa Y, Toyoki H, Sakaguchi H, Sato E and Tamaya T: Estrogen-related receptor expression in placenta throughout gestation. J Steroid Biochem Mol Biol. 94:67–69. 2005. View Article : Google Scholar
20	Hoffmann MJ, Florl AR, Seifert HH and Schulz WA: Multiple mechanisms downregulate CDKN1C in human bladder cancer. Int J Cancer. 114:406–413. 2005. View Article : Google Scholar
21	Thamizhanambi TP, Rameshkumar A, Ramya R, Krishnan R, Dineshkumar T and Nandhini G: Analysis of metabolic regulators PGC1-α and PGC1-β in oral squamous cell carcinoma with and without hyperglycemia. Asian Pac J Cancer Prev. 23:2797–2803. 2022. View Article : Google Scholar
22	Jafari N, Zargar SJ, Yassa N and Delnavazi MR: Induction of apoptosis and cell cycle arrest by Dorema glabrum root extracts in a gastric adenocarcinoma (AGS) cell line. Asian Pac J Cancer Prev. 17:5189–5193. 2016.
23	Piri-Gharaghie T, Ghajari G, Hassanpoor M, Jegargoshe-Shirin N, Soosanirad M, Khayati S, Farhadi-Biregani A and Mirzaei A: Investigation of antibacterial and anticancer effects of novel niosomal formulated Persian Gulf Sea cucumber extracts. Heliyon. 9:e141492023. View Article : Google Scholar
24	Zhang W, Shang X, Yang F, Han W, Xia H, Lu N, Liu Y and Wang X: CDC25C as a predictive biomarker for immune checkpoint inhibitors in patients with lung adenocarcinoma. Front Oncol. 12:8677882022. View Article : Google Scholar
25	Chen M, Wu R, Li G, Liu C, Tan L, Xiao K, Ye Y and Qin Z: Motor neuron and pancreas homeobox 1/HLXB9 promotes sustained proliferation in bladder cancer by upregulating CCNE1/2. J Exp Clin Cancer Res. 37:1542018. View Article : Google Scholar
26	Du Y, Zheng Y, Yu K, Zhan C and Qiao T: Genome-wide analyses of lung cancer after single high-dose radiation at five time points (2, 6, 12, 24, and 48 h). Front Genet. 14:11262362023. View Article : Google Scholar
27	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
28	Nanashima N, Horie K, Chiba M, Nakano M, Maeda H and Nakamura T: Anthocyanin-rich blackcurrant extract inhibits proliferation of the MCF10A healthy human breast epithelial cell line through induction of G0/G1 arrest and apoptosis. Mol Med Rep. 16:6134–6141. 2017. View Article : Google Scholar
29	Perdiguero E and Nebreda AR: Regulation of Cdc25C activity during the meiotic G2/M transition. Cell Cycle. 3:733–737. 2004. View Article : Google Scholar
30	Fagundes R and Teixeira LK: Cyclin E/CDK2: DNA replication, replication stress and genomic instability. Front Cell Dev Biol. 9:7748452021. View Article : Google Scholar
31	Hayward D, Alfonso-Pérez T and Gruneberg U: Orchestration of the spindle assembly checkpoint by CDK1-cyclin B1. FEBS Lett. 593:2889–2907. 2019. View Article : Google Scholar
32	Matsuoka S, Edwards MC, Bai C, Parker S, Zhang P, Baldini A, Harper JW and Elledge SJ: p57KIP2, a structurally distinct member of the p21CIP1 Cdk inhibitor family, is a candidate tumor suppressor gene. Genes Dev. 9:650–662. 1995. View Article : Google Scholar
33	Li FN, Zhang QY, Li O, Liu SL, Yang ZY, Pan LJ, Zhao C, Gong W, Shu YJ and Dong P: ESRRA promotes gastric cancer development by regulating the CDC25C/CDK1/CyclinB1 pathway via DSN1. Int J Biol Sci. 17:1909–1924. 2021. View Article : Google Scholar
34	Bedner E, Li X, Gorczyca W, Melamed MR and Darzynkiewicz Z: Analysis of apoptosis by laser scanning cytometry. Cytometry. 35:181–195. 1999. View Article : Google Scholar
35	Orlinick JR and Chao MV: TNF-related ligands and their receptors. Cell Signal. 10:543–551. 1998. View Article : Google Scholar
36	Nagata S and Golstein P: The Fas death factor. Science. 267:1449–1456. 1995. View Article : Google Scholar
37	Akagi T, Yoshino T and Kondo E: The Fas antigen and Fas-mediated apoptosis in B-cell differentiation. Leuk Lymphoma. 28:483–489. 1998. View Article : Google Scholar
38	Götz C and Montenarh M: p53: DNA damage, DNA repair, and apoptosis. Rev Physiol Biochem Pharmacol. 127:65–95. 1996. View Article : Google Scholar
39	De Vitto H, Ryu J, Calderon-Aparicio A, Monts J, Dey R, Chakraborty A, Lee MH, Bode AM and Dong Z: Estrogen-related receptor alpha directly binds to p53 and cooperatively controls colon cancer growth through the regulation of mitochondrial biogenesis and function. Cancer Metab. 8:282020. View Article : Google Scholar
40	Matsushima H, Mori T, Ito F, Yamamoto T, Akiyama M, Kokabu T, Yoriki K, Umemura S, Akashi K and Kitawaki J: Anti-tumor effect of estrogen-related receptor alpha knockdown on uterine endometrial cancer. Oncotarget. 7:34131–34148. 2016. View Article : Google Scholar
41	Sun P, Mao X, Gao M, Huang MM, Chen LL, Ruan GY, Huang WY, Braicu EI and Sehouli J: Novel endocrine therapeutic strategy in endometrial carcinoma targeting estrogen-related receptor α by XCT790 and siRNA. Cancer Manag Res. 10:2521–2535. 2018. View Article : Google Scholar
42	Zhang J, Guan X, Liang N and Li S: Estrogen-related receptor alpha triggers the proliferation and migration of human non-small cell lung cancer via interleukin-6. Cell Biochem Funct. 36:255–262. 2018. View Article : Google Scholar
43	Scarpulla RC: Metabolic control of mitochondrial biogenesis through the PGC-1 family regulatory network. Biochim Biophys Acta. 1813:1269–1278. 2011. View Article : Google Scholar
44	Schreiber SN, Knutti D, Brogli K, Uhlmann T and Kralli A: The transcriptional coactivator PGC-1 regulates the expression and activity of the orphan nuclear receptor estrogen-related receptor alpha (ERRalpha). J Biol Chem. 278:9013–9018. 2003. View Article : Google Scholar
45	Dufour CR, Wilson BJ, Huss JM, Kelly DP, Alaynick WA, Downes M, Evans RM, Blanchette M and Giguère V: Genome-wide orchestration of cardiac functions by the orphan nuclear receptors ERRalpha and gamma. Cell Metab. 5:345–356. 2007. View Article : Google Scholar
46	Suetsugi M, Su L, Karlsberg K, Yuan YC and Chen S: Flavone and isoflavone phytoestrogens are agonists of estrogen-related receptors. Mol Cancer Res. 1:981–991. 2003.