Sox9 is a β-catenin-regulated transcription factor that enhances the colony-forming activity of squamous cell carcinoma cells

Li,Xue  Mei; Piao,Yong  Jun; Sohn,Kyung‑Cheol; Ha,Jeong-Min; Im,Myung; Seo,Young‑Joon; Whang,Kyu  Uang; Lee,Jeung‑Hoon; Lee,Young; Kim,Chang  Deok

doi:10.3892/mmr.2016.5210

Sox9 is a β-catenin-regulated transcription factor that enhances the colony-forming activity of squamous cell carcinoma cells

Authors:
- Xue Mei Li
- Yong Jun Piao
- Kyung‑Cheol Sohn
- Jeong-Min Ha
- Myung Im
- Young‑Joon Seo
- Kyu Uang Whang
- Jeung‑Hoon Lee
- Young Lee
- Chang Deok Kim
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Affiliations: Department of Dermatology, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, P.R. China, Department of Dermatology, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116044, P.R. China, Department of Dermatology, School of Medicine, Chungnam National University, Daejeon 301‑747, Republic of Korea, Department of Dermatology, College of Medicine, Soonchunhyang University, Seoul 330‑721, Republic of Korea
Published online on: May 4, 2016 https://doi.org/10.3892/mmr.2016.5210
Pages: 337-342

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Abstract

Squamous cell carcinoma (SCC) is a common skin cancer, of which the incidence is relatively high, ranking second among the non‑melanoma skin cancers. It is known that numerous intracellular signal regulators are involved in the pathogenesis of SCC. The Wnt/β-catenin signaling pathway serves an important role in cancer development. However, the downstream effectors of β‑catenin remain to be clearly elucidated yet. The present study investigated the functional importance of Wnt/β‑catenin signaling in cutaneous SCC. β‑catenin expression was reduced using recombinant adenovirus expressing specific microRNA (miR). Knockdown of β‑catenin resulted in a marked reduction of the colony-forming activity of the SCC cells, SCC12. In an attempt to identify the β‑catenin downstream genes, it was found that Sox9 was regulated by β‑catenin in SCC12 cells. Overexpression of a constitutively active form of β‑catenin led to the induction of Sox9, while knockdown of β‑catenin resulted in downregulation of Sox9. When the expression of Sox9 was reduced using specific miR, colony-forming activity of the SCC12 cells was significantly reduced. When Sox9 was overexpressed in cells where β‑catenin was knocked down, it partially restored the colony‑forming potential. Taken together, the present results suggested that Sox9 is a β-catenin downstream transcription factor and is positively involved in SCC development.

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1	Alam M and Ratner D: Cutaneous squamous-cell carcinoma. N Engl J Med. 344:975–983. 2001. View Article : Google Scholar : PubMed/NCBI
2	Brash DE, Rudolph JA, Simon JA, Lin A, McKenna GJ, Baden HP, Halperin AJ and Pontén J: A role for sunlight in skin cancer: UV-induced p53 mutations in squamous cell carcinoma. Proc Natl Acad Sci USA. 88:10124–10128. 1991. View Article : Google Scholar : PubMed/NCBI
3	Kolev V, Mandinova A, Guinea-Viniegra J, Hu B, Lefort K, Lambertini C, Neel V, Dummer R, Wagner EF and Dotto GP: EGFR signalling as a negative regulator of Notch1 gene transcription and function in proliferating keratinocytes and cancer. Nat Cell Biol. 10:902–911. 2008. View Article : Google Scholar : PubMed/NCBI
4	van Hogerlinden M, Rozell BL, Ahrlund-Richter L and Toftgård R: Squamous cell carcinomas and increased apoptosis in skin with inhibited Rel/nuclear factor-kappaB signaling. Cancer Res. 59:3299–3303. 1999.PubMed/NCBI
5	Brasanac D, Boricic I, Todorovic V, Tomanovic N and Radojevic S: Cyclin A and beta-catenin expression in actinic keratosis, Bowen's disease and invasive squamous cell carcinoma of the skin. Br J Dermatol. 153:1166–1175. 2005. View Article : Google Scholar : PubMed/NCBI
6	Lyakhovitsky A, Barzilai A, Fogel M, Trau H and Huszar M: Expression of e-cadherin and beta-catenin in cutaneous squamous cell carcinoma and its precursors. Am J Dermatopathol. 26:372–378. 2004. View Article : Google Scholar : PubMed/NCBI
7	Doglioni C, Piccinin S, Demontis S, Cangi MG, Pecciarini L, Chiarelli C, Armellin M, Vukosavljevic T, Boiocchi M and Maestro R: Alterations of beta-catenin pathway in non-melanoma skin tumors: Loss of alpha-ABC nuclear reactivity correlates with the presence of beta-catenin gene mutation. Am J Pathol. 163:2277–2287. 2003. View Article : Google Scholar : PubMed/NCBI
8	Hisken J and Behrens J: The Wnt signaling pathway. J Cell Sci. 113:3545–3546. 2000.
9	Zhang Y, Liu B, Zhao Q, Hou T and Huang X: Nuclear local-izaiton of β-catenin is associated with poor survival and chemo-/radioresistance in human cervical squamous cell cancer. Int J Clin Exp Pathol. 7:3908–3917. 2014.
10	Li P, Cao Y, Li Y, Zhou L, Liu X and Geng M: Expression of Wnt-5a and β-catenin in primary hepatocellular carcinoma. Int J Clin Exp Pathol. 7:3190–3195. 2014.
11	Cui J, Xi H, Cai A, Bian S, Wei B and Chen L: Decreased expression of Sox7 correlates with the upregulation of the Wnt/β-catenin signaling pathway and the poor survival of gastric cancer patients. Int J Mol Med. 34:197–204. 2014.PubMed/NCBI
12	Malanchi I, Peinado H, Kassen D, Hussenet T, Metzger D, Chambon P, Huber M, Hohl D, Cano A, Birchmeier W and Huelsken J: Cutaneous cancer stem cell maintenance is dependent on beta-catenin signalling. Nature. 452:650–653. 2008. View Article : Google Scholar : PubMed/NCBI
13	Popp S, Waltering S, Herbst C, Moll I and Boukamp P: UV-B-type mutations and chromosomal imbalances indicate common pathways for the development of Merkel and skin squamous cell carcinomas. Int J Cancer. 99:352–360. 2002. View Article : Google Scholar : PubMed/NCBI
14	Choi HI, Sohn KC, Hong DK, Lee Y, Kim CD, Yoon TJ, Park JW, Jung S, Lee JH and Lee YH: Melanosome uptake is associated with the proliferation and differentiation of keratinocytes. Arch Dermatol Res. 306:59–66. 2014. View Article : Google Scholar
15	Je YJ, Choi DK, Sohn KC, Kim HR, Im M, Lee Y, Lee JH, Kim CD and Seo YJ: Inhibitory role of Id1 on TGF-β-induced collagen expression in human dermal fibroblasts. Biochem Biophys Res Commun. 444:81–85. 2014. View Article : Google Scholar : PubMed/NCBI
16	Lee JS, Kim DH, Choi DK, Kim CD, Ahn GB, Yoon TY, Lee JH and Lee JY: Comparison of gene expression profiles between keratinocytes, melanocytes and fibroblasts. Ann Dermatol. 25:36–45. 2013. View Article : Google Scholar : PubMed/NCBI
17	Sohn KC, Shi G, Jang S, Choi DK, Lee Y, Yoon TJ, Park H, Hwang C, Kim HJ, Seo YJ, et al: Pitx2, a beta-catenin-regulated transcription factor, regulates the differentiation of outer root sheath cells cultured in vitro. J Dermatol Sci. 54:6–11. 2009. View Article : Google Scholar : PubMed/NCBI
18	Shi G, Sohn KC, Li Z, Choi DK, Park YM, Kim JH, Fan YM, Nam YH, Kim S, Im M, et al: Expression and functional role of Sox9 in human epidermal keratinocytes. PLoS One. 8:e543552013. View Article : Google Scholar : PubMed/NCBI
19	Franken NA, Rodermond HM, Stap J, Haveman J and van Bree C: Clonogenic assay of cells in vitro. Nat Protoc. 1:2315–2319. 2006. View Article : Google Scholar
20	Gat U, DasGupta R, Degenstein L and Fuchs E: De Novo hair follicle morphogenesis and hair tumors in mice expressing a truncated beta-catenin in skin. Cell. 95:605–614. 1998. View Article : Google Scholar : PubMed/NCBI
21	Lefebvre V, Dumitriu B, Penzo-Méndez A, Han Y and Pallavi B: Control of cell fate and differentiation by Sry-related high-mobility-group box (Sox) transcription factors. Int J Biochem Cell Biol. 39:2195–2214. 2007. View Article : Google Scholar : PubMed/NCBI
22	Sarkar A and Hochedlinger K: The sox family of transcription factors: Versatile regulators of stem and progenitor cell fate. Cell Stem Cell. 12:15–30. 2013. View Article : Google Scholar : PubMed/NCBI
23	Foster JW, Dominguez-Steglich MA, Guioli S, Kwok C, Weller PA, Stevanović M, Weissenbach J, Mansour S, Young ID, Goodfellow PN, et al: Campomelic dysplasia and autosomal sex reversal caused by mutations in an SRY-related gene. Nature. 372:525–530. 1994. View Article : Google Scholar : PubMed/NCBI
24	Schafer AJ, Foster JW, Kwok C, Weller PA, Guioli S and Goodfellow PN: Campomelic dysplasia with XY sex reversal: Diverse phenotypes resulting from mutations in a single gene. Ann NY Acad Sci. 785:137–149. 1996. View Article : Google Scholar : PubMed/NCBI
25	Chakravarty G, Moroz K, Makridakis NM, Lloyd SA, Galvez SE, Canavello PR, Lacey MR, Agrawal K and Mondal D: Prognostic significance of cytoplasmic SOX9 in invasive ductal carcinoma and metastatic breast cancer. Exp Biol Med (Maywood). 236:145–155. 2011. View Article : Google Scholar
26	Wang H, He L, Ma F, Regan MM, Balk SP, Richardson AL and Yuan X: SOX9 regulates low density lipoprotein receptor-related protein 6 (LRP6) and T-cell factor 4 (TCF4) expression and Wnt/β-catenin activation in breast cancer. J Biol Chem. 288:6478–6487. 2013. View Article : Google Scholar : PubMed/NCBI
27	Matheu A, Collado M, Wise C, Manterola L, Cekaite L, Tye AJ, Canamero M, Bujanda L, Schedl A, Cheah KS, et al: Oncogenicity of the developmental transcription factor Sox9. Cancer Res. 72:1301–1315. 2012. View Article : Google Scholar : PubMed/NCBI
28	Panza A, Pazienza V, Ripoli M, Benegiamo G, Gentile A, Valvano MR, Augello B, Merla G, Prattichizzo C, Tavano F, et al: Interplay between SOX9, β-catenin and PPARγ activation in colorectal cancer. Biochim Biophys Acta. 1833:1853–1865. 2013. View Article : Google Scholar : PubMed/NCBI
29	Yano F, Kugimiya F, Ohba S, Ikeda T, Chikuda H, Ogasawara T, Ogata N, Takato T, Nakamura K, Kawaguchi H and Chung UI: The canonical Wnt signaling pathway promotes chondrocyte differentiation in a Sox9-dependent manner. Biochem Biophys Res Commun. 333:1300–1308. 2005. View Article : Google Scholar : PubMed/NCBI
30	Blache P, van de Wetering M, Duluc I, Domon C, Berta P, Freund JN, Clevers H and Jay P: SOX9 is an intestine crypt transcription factor, is regulated by the Wnt pathway, and represses the CDX2 and MUC2 genes. J Cell Biol. 166:37–47. 2004. View Article : Google Scholar : PubMed/NCBI
31	Feng Y, Sentani K, Wiese A, Sands E, Green M, Bommer GT, Cho KR and Fearon ER: Sox9 induction, ectopic Paneth cells, and mitotic spindle axis defects in mouse colon adenomatous epithelium arising from conditional biallelic Apc inactivation. Am J Pathol. 183:493–503. 2013. View Article : Google Scholar : PubMed/NCBI