Cytoplasmic expression of SOX9 as a poor prognostic factor for oral squamous cell carcinoma

Sumita,Yoshimasa; Yamazaki,Manabu; Maruyama,Satoshi; Abé,Tatsuya; Cheng,Jun; Takagi,Ritsuo; Tanuma,Jun‑Ichi

doi:10.3892/or.2018.6665

Cytoplasmic expression of SOX9 as a poor prognostic factor for oral squamous cell carcinoma

Authors:
- Yoshimasa Sumita
- Manabu Yamazaki
- Satoshi Maruyama
- Tatsuya Abé
- Jun Cheng
- Ritsuo Takagi
- Jun‑Ichi Tanuma
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Affiliations: Division of Oral Pathology, Niigata University Graduate School of Medical and Dental Sciences, Niigata 951‑8514, Japan, Oral Pathology Section, Department of Surgical Pathology, Niigata University Hospital, Niigata 951‑8520, Japan, Division of Molecular and Diagnostic Pathology, Niigata University Graduate School of Medical and Dental Sciences, Niigata 951‑8122, Japan, Division of Oral and Maxillofacial Surgery, Niigata University Graduate School of Medical and Dental Sciences, Niigata 951‑8514, Japan
Published online on: August 22, 2018 https://doi.org/10.3892/or.2018.6665
Pages: 2487-2496
Copyright: © Sumita et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Transcription factor SRY‑box 9 (SOX9) is a key regulator of chondrocyte differentiation and sex determination, and it is also involved in the progression of various types of human cancer. However, its putative association with oral squamous cell carcinoma (OSCC) remains elusive. The aim of the present study was to investigate the expression profiles of SOX9 in various oral epithelial lesions, including OSCC. We performed immunohistochemical analysis of SOX9 expression in surgical specimens of OSCC, which simultaneously exhibited different grades of epithelial lesions, and analyzed the correlation between SOX9 expression and several clinicopathological factors. Moreover, we performed immunofluorescent staining, western blot analysis and real‑time reverse transcription‑polymerase chain reaction to assess SOX9 expression in OSCC HSC‑3 (a metastatic cell line) and HSC‑4 (a non‑metastatic cell line) cell lines. In surgical specimens, SOX9 expression was detected in the nuclei of proliferating cells in areas with epithelial dysplasia and carcinoma in situ, but not in areas with normal epithelia. Nuclear SOX9 expression was observed in most SCC cells. Notably, cytoplasmic SOX9 expression was confirmed only in some SCC cells; however, cytoplasmic SOX9 expression was significantly and positively correlated with poor clinical outcomes. Both protein and mRNA expression of SOX9 were significantly higher in the HSC‑3 cell line than that in the HSC‑4 line. Notably, however, only HSC‑3 cells exhibited cytoplasmic localization of SOX9 expression. Our findings indicate that SOX9 may be involved in the tumorigenesis and progression of OSCC. Furthermore, its cytoplasmic expression represents a potential predictive biomarker for tumor aggressiveness and OSCC prognosis.

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1	Jo A, Denduluri S, Zhang B, Wang Z, Yin L, Yan Z, Kang R, Shi LL, Mok J, Lee MJ and Haydon RC: The versatile functions of Sox9 in development, stem cells, and human diseases. Genes Dis. 1:149–161. 2014. View Article : Google Scholar : PubMed/NCBI
2	Pritchett J, Athwal V, Roberts N, Hanley NA and Hanley KP: Understanding the role of SOX9 in acquired diseases: Lessons from development. Trends Mol Med. 17:166–174. 2011. View Article : Google Scholar : PubMed/NCBI
3	Harley VR, Clarkson MJ and Argentaro A: The molecular action and regulation of the testis-determining factors, SRY (sex-determining region on the Y chromosome) and SOX9 [SRY-related high-mobility group (HMG) box 9]. Endocr Rev. 24:466–487. 2003. View Article : Google Scholar : PubMed/NCBI
4	Akiyama H: Control of chondrogenesis by the transcription factor Sox9. Mod Rheumatol. 18:213–219. 2008. View Article : Google Scholar : PubMed/NCBI
5	Vidal VP, Chaboissier MC, Lutzkendorf S, Cotsarelis G, Mill P, Hui CC, Ortonne N, Ortonne JP and Schedl A: Sox9 is essential for outer root sheath differentiation and the formation of the hair stem cell compartment. Curr Biol. 15:1340–1351. 2005. View Article : Google Scholar : PubMed/NCBI
6	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
7	Wang H, McKnight NC, Zhang T, Lu ML, Balk SP and Yuan X: SOX9 is expressed in normal prostate basal cells and regulates androgen receptor expression in prostate cancer cells. Cancer Res. 67:528–536. 2007. View Article : Google Scholar : PubMed/NCBI
8	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. 236:145–155. 2011. View Article : Google Scholar
9	Lü B, Fang Y, Xu J, Wang L, Xu F, Xu E, Huang Q and Lai M: Analysis of SOX9 expression in colorectal cancer. Am J Clin Pathol. 130:897–904. 2008. View Article : Google Scholar : PubMed/NCBI
10	Vidal VP, Ortonne N and Schedl A: SOX9 expression is a general marker of basal cell carcinoma and adnexal-related neoplasms. J Cutan Pathol. 35:373–379. 2008. View Article : Google Scholar : PubMed/NCBI
11	Hong Y, Chen W, Du X, Ning H, Chen H, Shi R, Lin S, Xu R, Zhu J, Wu S, et al: Upregulation of sex-determining region Y-box 9 (SOX9) promotes cell proliferation and tumorigenicity in esophageal squamous cell carcinoma. Oncotarget. 6:31241–31254. 2015. View Article : Google Scholar : PubMed/NCBI
12	Ikarashi T, Ida-Yonemochi H, Ohshiro K, Cheng J and Saku T: Intraepithelial expression of perlecan, a basement membrane-type heparan sulfate proteoglycan reflects dysplastic changes of the oral mucosal epithelium. J Oral Pathol Med. 33:87–95. 2004. View Article : Google Scholar : PubMed/NCBI
13	Maruyama S, Shimazu Y, Kudo T, Sato K, Yamazaki M, Abé T, Babkair H, Cheng J, Aoba T and Saku T: Three-dimensional visualization of perlecan-rich neoplastic stroma induced concurrently with the invasion of oral squamous cell carcinoma. J Oral Pathol Med. 43:627–636. 2014. View Article : Google Scholar : PubMed/NCBI
14	Maruyama S, Cheng J, Yamazaki M, Liu A and Saku T: Keratinocyte growth factor colocalized with perlecan at the site of capsular invasion and vascular involvement in salivary pleomorphic adenomas. J Oral Pathol Med. 38:377–385. 2009. View Article : Google Scholar : PubMed/NCBI
15	Toyoshima K, Kimura S, Cheng J, Oda Y, Mori KJ and Saku T: High-molecular-weight fibronectin synthesized by adenoid cystic carcinoma cells of salivary gland origin. Jpn J Cancer Res. 90:308–319. 1999. View Article : Google Scholar : PubMed/NCBI
16	Hasegawa M, Cheng J, Maruyama S, Yamazaki M, Abé T, Babkair H, Saito C and Saku T: Differential immunohistochemical expression profiles of perlecan-binding growth factors in epithelial dysplasia, carcinoma in situ, and squamous cell carcinoma of the oral mucosa. Pathol Res Pract. 212:426–436. 2016. View Article : Google Scholar : PubMed/NCBI
17	Warren CR, Grindel BJ, Francis L, Carson DD and Farach-Carson MC: Transcriptional activation by NFkappaB increases perlecan/HSPG2 expression in the desmoplastic prostate tumor microenvironment. J Cell Biochem. 115:1322–1333. 2014. View Article : Google Scholar : PubMed/NCBI
18	Larsimont JC, Youssef KK, Sánchez-Danés A, Sukumaran V, Defrance M, Delatte B, Liagre M, Baatsen P, Marine JC, Lippens S, et al: Sox9 controls self-renewal of oncogene targeted cells and links tumor initiation and invasion. Cell Stem Cell. 17:60–73. 2015. View Article : Google Scholar : PubMed/NCBI
19	UICC International Union Against Cancer: TNM Classification of Malignant Tumours. Sobin LH, Gospodarowicz MK and Wittekind C: 7th edition. Wiley-Blackwell; Hoboken, NJ: pp. 25–28. 2009
20	Saku T and Furthmayr H: Characterization of the major heparan sulfate proteoglycan secreted by bovine aortic endothelial cells in culture. Homology to the large molecular weight molecule of basement membranes. J Biol Chem. 264:3514–3523. 1989.PubMed/NCBI
21	Momose F, Araida T, Negishi A, Ichijo H, Shioda S and Sasaki S: Variant sublines with different metastatic potentials selected in nude mice from human oral squamous cell carcinomas. J Oral Pathol Med. 18:391–395. 1989. View Article : Google Scholar : PubMed/NCBI
22	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2^−ΔΔCT method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
23	Yamazaki M, Maruyama S, Abé T, Essa A, Babkair H, Cheng J and Saku T: MFG-E8 expression for progression of oral squamous cell carcinoma and for self-clearance of apoptotic cells. Lab Invest. 94:1260–1272. 2014. View Article : Google Scholar : PubMed/NCBI
24	Ahsan MS, Yamazaki M, Maruyama S, Kobayashi T, Ida-Yonemochi H, Hasegawa M, Ademola Henry A, Cheng J and Saku T: Differential expression of perlecan receptors, α-dystroglycan and integrin β1, before and after invasion of oral squamous cell carcinoma. J Oral Pathol Med. 40:552–559. 2011. View Article : Google Scholar : PubMed/NCBI
25	Kobayashi T, Maruyama S, Cheng J, Ida-Yonemochi H, Yagi M, Takagi R and Saku T: Histopathological varieties of oral carcinoma in situ: Diagnosis aided by immunohistochemistry dealing with the second basal cell layer as the proliferating center of oral mucosal epithelia. Pathol Int. 60:156–166. 2010. View Article : Google Scholar : PubMed/NCBI
26	Mikami T, Cheng J, Maruyama S, Kobayashi T, Funayama A, Yamazaki M, Adeola HA, Wu L, Shingaki S, Saito C and Saku T: Emergence of keratin 17 vs. loss of keratin 13: Their reciprocal immunohistochemical profiles in oral carcinoma in situ. Oral Oncol. 47:497–503. 2011. View Article : Google Scholar : PubMed/NCBI
27	Jiang SS, Fang WT, Hou YH, Huang SF, Yen BL, Chang JL, Li SM, Liu HP, Liu YL, Huang CT, et al: Upregulation of SOX9 in lung adenocarcinoma and its involvement in the regulation of cell growth and tumorigenicity. Clin Cancer Res. 16:4363–4373. 2010. View Article : Google Scholar : PubMed/NCBI
28	Alvarado CG, Maruyama S, Cheng J, Ida-Yonemochi H, Kobayashi T, Yamazaki M, Takagi R and Saku T: Nuclear translocation of β-catenin synchronized with loss of E-cadherin in oral epithelial dysplasia with a characteristic two-phase appearance. Histopathology. 59:283–291. 2011. View Article : Google Scholar : PubMed/NCBI
29	Malki S, Berta P, Poulat F and Boizet-Bonhoure B: Cytoplasmic retention of the sex-determining factor SOX9 via the microtubule network. Exp Cell Res. 309:468–475. 2005. View Article : Google Scholar : PubMed/NCBI
30	Oz Bar M, Kumar A, Elayyan J, Reich E, Binyamin M, Kandel L, Liebergall M, Steinmeyer J, Lefebvre V and Dvir-Ginzberg M: Acetylation reduces SOX9 nuclear entry and ACAN gene transactivation in human chondrocytes. Aging Cell. 15:499–508. 2016. View Article : Google Scholar : PubMed/NCBI
31	Sim H, Argentaro A and Harley VR: Boys, girls and shuttling of SRY and SOX9. Trends Endocrinol Metab. 19:213–222. 2008. View Article : Google Scholar : PubMed/NCBI
32	Guo W, Keckesova Z, Donaher JL, Shibue T, Tischler V, Reinhardt F, Itzkovitz S, Noske A, Zürrer-Härdi U, Bell G, et al: Slug and Sox9 cooperatively determine the mammary stem cell state. Cell. 148:1015–1028. 2012. View Article : Google Scholar : PubMed/NCBI
33	Fazilaty H, Gardaneh M, Akbari P, Zekri A and Behnam B: SLUG and SOX9 cooperatively regulate tumor initiating niche factors in breast cancer. Cancer Microenviron. 9:71–74. 2016. View Article : Google Scholar : PubMed/NCBI
34	Savagner P, Yamada KM and Thiery JP: The zinc-finger protein slug causes desmosome dissociation, an initial and necessary step for growth factor-induced epithelial-mesenchymal transition. J Cell Biol. 137:1403–1419. 1997. View Article : Google Scholar : PubMed/NCBI
35	Sadatsuki R, Kaneko H, Kinoshita M, Futami I, Nonaka R, Culley KL, Otero M, Hada S, Goldring MB, Yamada Y, et al: Perlecan is required for the chondrogenic differentiation of synovial mesenchymal cells through regulation of Sox9 gene expression. J Orthop Res. 35:837–846. 2017. View Article : Google Scholar : PubMed/NCBI
36	Khanom R, Nguyen CT, Kayamori K, Zhao X, Morita K, Miki Y, Katsube K, Yamaguchi A and Sakamoto K: Keratin 17 is induced in oral cancer and facilitates tumor growth. PLoS One. 11:e01611632016. View Article : Google Scholar : PubMed/NCBI