Effect of Hath1 on the proliferation and apoptosis of cutaneous squamous cell carcinoma in vitro

Ying,Zuolin; Li,Xiaojie; Dang,Hong; Wang,Feng; Xu,Xiaoyan

doi:10.3892/mmr.2015.4463

Effect of Hath1 on the proliferation and apoptosis of cutaneous squamous cell carcinoma in vitro

Authors:
- Zuolin Ying
- Xiaojie Li
- Hong Dang
- Feng Wang
- Xiaoyan Xu
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Affiliations: Department of Dermatology, Shanghai First People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200080, P.R. China, Experimental Research Center, Shanghai First People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200080, P.R. China
Published online on: October 21, 2015 https://doi.org/10.3892/mmr.2015.4463
Pages: 7845-7850
Copyright: © Ying et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Increasing evidence has demonstrated that the tumor suppressor gene Hath1 is implicated in the development and progression of tumors and is verified to be downregulated in several types of tumor. However, the roles and precise molecular mechanisms of Hath1 in cutaneous squamous cell carcinoma (SCC) remain to be elucidated. In the present study, two approaches were used to investigate the tumor‑suppressing effect of Hath1 in cutaneous SCC. Firstly, the effect of inhibiting Hath1 expression with short hairpin RNA (shRNA) on tumor growth and apoptosis was investigated. KUMA5 cells were stably transfected with a plasmid expressing Hath1 shRNA (pGenesil‑1‑Hath1). Secondly, the anti‑tumor effect of Hath1 was investigated in KUMA5 cells following transfection with pcDNA3.1‑Hath1. The mRNA and protein expression of Hath1 was detected by reverse transcription quantitative polymerase chain reaction and western blot analysis, respectively. Cell proliferation in vitro was assessed using an MTT assay. Flow cytometry was used to detect cell apoptosis. The results demonstrated that compared with the control groups, the expression of Hath1 was significantly reduced in the KUMA5/pGenesil‑1‑Hath1 cells and markedly increased in the KUMA5/pcDNA3.1‑Hath1 cells. Cell proliferation was markedly increased in the KUMA5/pGenesil‑1‑Hath1 cells in a time‑dependent manner; however, it was markedly inhibited in the KUMA5/pcDNA3.1‑Hath1 cells. Flow cytometry revealed that apoptosis decreased in KUMA5/pGenesil‑1‑Hath1 cells and increased in KUMA5/pcDNA3.1‑Hath1 cells. Downregulation of Hath1 expression promoted the proliferation and reduced the apoptosis of KUMA5 cells. By contrast, overexpression of Hath1 inhibited proliferation and induced the apoptosis of KUMA5 cells. These findings provide possible new strategies and therapeutic targets for the treatment and diagnosis of cutaneous SCC.

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1	Veness MJ, Morgan GJ, Palme CE and Gebski V: Surgery and adjuvant radiotherapy in patients with cutaneous head and neck squamous cell carcinoma metastatic to lymph nodes: Combined treatment should be considered best practice. Laryngoscope. 115:870–875. 2005. View Article : Google Scholar : PubMed/NCBI
2	Preston DS and Stern RS: Nonmelanoma cancers of the skin. N Engl J Med. 327:1649–1662. 1992. View Article : Google Scholar : PubMed/NCBI
3	Euvrard S, Kanitakis J and Claudy A: Skin cancers after organ transplantation. N Engl J Med. 348:1681–1691. 2003. View Article : Google Scholar : PubMed/NCBI
4	McGuire JF, Ge NN and Dyson S: Nonmelanoma skin cancer of the head and neck I: Histopathology and clinical behavior. Am J Otolaryngol. 30:121–133. 2009. View Article : Google Scholar : PubMed/NCBI
5	Martinez JC and Cook JL: High-risk cutaneous squamous cell carcinoma without palpable lymphadenopathy: Is there a therapeutic role for elective neck dissection? Dermatol Surg. 33:410–420. 2007.PubMed/NCBI
6	Weinberg AS, Ogle CA and Shim EK: Metastatic cutaneous squamous cell carcinoma: An update. Dermatol Surg. 33:885–899. 2007.PubMed/NCBI
7	Bertero T, Bourget-Ponzio I, Puissant A, et al: Tumor suppressor function of miR-483-3p on squamous cell carcinomas due to its pro-apoptotic properties. Cell Cycle. 12:2183–2193. 2013. View Article : Google Scholar : PubMed/NCBI
8	Liu D, Feng X, Wu X, et al: Tumor suppressor in lung cancer 1 (TSLC1), a novel tumor suppressor gene, is implicated in the regulation of proliferation, invasion, cell cycle, apoptosis and tumorigenicity in cutaneous squamous cell carcinoma. Tumour Biol. 34:3773–3783. 2013. View Article : Google Scholar : PubMed/NCBI
9	Yang C, Wu D, Jia J, et al: DLC1 as a regulator of proliferation, invasion, cell cycle and apoptosis in cutaneous squamous cell carcinoma. Tumour Biol. 34:2633–2643. 2013. View Article : Google Scholar : PubMed/NCBI
10	Scortegagna M, Martin RJ, Kladney RD, Neumann RG and Arbeit JM: Hypoxia-inducible factor-1alpha suppresses squamous carcinogenic progression and epithelial-mesenchymal transition. Cancer Res. 69:2638–2646. 2009. View Article : Google Scholar : PubMed/NCBI
11	Sharma A, Ramanjaneyulu A, Ray R and Rajeswari MR: Involvement of high mobility group B proteins in cisplatin-induced cytotoxicity in squamous cell carcinoma of skin. DNA Cell Biol. 28:311–318. 2009. View Article : Google Scholar : PubMed/NCBI
12	Bito T, Sumita N, Ashida M, et al: Inhibition of epidermal growth factor receptor and PI3K/Akt signaling suppresses cell proliferation and survival through regulation of Stat3 activation in human cutaneous squamous cell carcinoma. J Skin Cancer. 2011:8745712011. View Article : Google Scholar : PubMed/NCBI
13	Ben-Arie N, McCall AE, Berkman S, Eichele G, Bellen HJ and Zoghbi HY: Evolutionary conservation of sequence and expression of the bHLH protein Atonal suggests a conserved role in neurogenesis. Hum Mol Genet. 5:1207–1216. 1996. View Article : Google Scholar : PubMed/NCBI
14	Leow CC, Romero MS, Ross S, Polakis P and Gao WQ: Hath1, down-regulated in colon adenocarcinomas, inhibits proliferation and tumorigenesis of colon cancer cells. Cancer Res. 64:6050–6057. 2004. View Article : Google Scholar : PubMed/NCBI
15	Leow CC, Polakis P and Gao WQ: A role for Hath1, a bHLH transcription factor, in colon adenocarcinoma. Ann NY Acad Sci. 1059:174–183. 2005. View Article : Google Scholar : PubMed/NCBI
16	Bossuyt W, Kazanjian A, De Geest N, et al: Atonal homolog 1 is a tumor suppressor gene. PLoS Biol. 7:e392009. View Article : Google Scholar : PubMed/NCBI
17	Sikandar SS, Pate KT, Anderson S, et al: NOTCH signaling is required for formation and self-renewal of tumor-initiating cells and for repression of secretory cell differentiation in colon cancer. Cancer Res. 70:1469–1478. 2010. View Article : Google Scholar : PubMed/NCBI
18	Kazanjian A, Noah T, Brown D, Burkart J and Shroyer NF: Atonal homolog 1 is required for growth and differentiation effects of notch/gamma-secretase inhibitors on normal and cancerous intestinal epithelial cells. Gastroenterology. 139:918–928. 2010. View Article : Google Scholar : PubMed/NCBI
19	Zhu DH, Gong J, Ren K, et al: Hath1 expression is down-regulated in colon non-mucinous adenocarcinomas compared with the expression in normal colon mucosa. Chong Qing Yi Ke Da Xue Xue Bao. 35:666–669. 2010.In Chinese.
20	Zhu DH, Gong JP, Ren K, Sun JM and Wei SD: Hath1 gene transfer inhibits the proliferation of colonic cancer cells in vitro. Nan Fang Yi Ke Da Xue Xue Bao. 30:1005–1008. 2010.In Chinese. PubMed/NCBI
21	Zhu DH, Wen YW, Yang H and Liang G: Anti-cancer effect of Hath1 in pathogenesis of non-mucinous colon adenocarcinoma. Ji Lin Da Xue Xue Bao (Yi Xue Ban). 37:700–706. 2011.In Chinese.
22	Zhao LL, Liu Z and Zhang YJ: Hath1 gene inhibits colorectal cancer cell growth. Zhong Guo Yi Yao Sheng Wu Ji Shu. 6:204–208. 2011.In Chinese.
23	Peignon G, Durand A, Cacheux W, et al: Complex interplay between β-catenin signalling and Notch effectors in intestinal tumorigenesis. Gut. 60:166–176. 2011. View Article : Google Scholar : PubMed/NCBI
24	Zhu DH, Niu BL, Du HM, Ren K, Sun JM and Gong JP: Hath1 inhibits proliferation of colon cancer cells probably through up-regulating expression of Muc2 and p27 and down-regulating expression of cyclin D1. Asian Pac J Cancer Prev. 13:6349–6355. 2012. View Article : Google Scholar : PubMed/NCBI
25	Zhao H, Ayrault O, Zindy F, Kim JH and Roussel MF: Post-transcriptional down-regulation of Atoh1/Math1 by bone morphogenic proteins suppresses medulloblastoma development. Genes Dev. 22:722–727. 2008. View Article : Google Scholar : PubMed/NCBI
26	Flora A, Klisch TJ, Schuster G and Zoghbi HY: Deletion of Atoh1 disrupts Sonic Hedgehog signaling in the developing cerebellum and prevents medulloblastoma. Science. 326:1424–1427. 2009. View Article : Google Scholar : PubMed/NCBI
27	Sekine A, Akiyama Y, Yanagihara K and Yuasa Y: Hath1 up-regulates gastric mucin gene expression in gastric cells. Biochem Biophys Res Commun. 344:1166–1171. 2006. View Article : Google Scholar : PubMed/NCBI
28	Zhang X, Yu H, Yang Y, et al: SOX2 in gastric carcinoma, but not Hath1, is related to patients' clinicopathological features and prognosis. J Gastrointest Surg. 14:1220–1226. 2010. View Article : Google Scholar : PubMed/NCBI
29	Heiskala K, Arola J, Heiskala M and Andersson LC: Expression of Reg IV and Hath1 in neuroendocrine neoplasms. Histol Histopathol. 25:63–72. 2010.
30	Chen JB, Liu JH, Liu YF, et al: Expression and clinical significance of Atoh1 in lung cancer. Lin Chuang Fei Ke Za Zhi. 11:1744–1746. 2011.In Chinese.
31	Xu HT, Xie XM, Li QC, et al: Atonal homolog 1 expression in lung cancer correlates with inhibitors of the Wnt pathway as well as the differentiation and primary tumor stage. APMIS. 121:111–119. 2013. View Article : Google Scholar
32	Leonard JH, Cook AL, Van Gele M, et al: Proneural and proneuroendocrine transcription factor expression in cutaneous mechanoreceptor (Merkel) cells and Merkel cell carcinoma. Int J Cancer. 101:103–110. 2002. View Article : Google Scholar : PubMed/NCBI
33	Ben-Arie N, Bellen HJ, Armstrong DL, et al: Math1 is essential for genesis of cerebellar granule neurons. Nature. 390:169–172. 1997. View Article : Google Scholar : PubMed/NCBI
34	Bermingham NA, Hassan BA, Price SD, et al: Math1: An essential gene for the generation of inner ear hair cells. Science. 284:1837–1841. 1999. View Article : Google Scholar : PubMed/NCBI
35	Yang Q, Bermingham NA, Finegold MJ and Zoghbi HY: Requirement of Math1 for secretory cell lineage commitment in the mouse intestine. Science. 294:2155–2158. 2001. View Article : Google Scholar : PubMed/NCBI
36	Shroyer NF, Helmrath MA, Wang VY, Antalffy B, Henning SJ and Zoghbi HY: Intestine-specific ablation of mouse atonal homolog 1 (Math1) reveals a role in cellular homeostasis. Gastroenterology. 132:2478–2488. 2007. View Article : Google Scholar : PubMed/NCBI
37	Rose MF, Ren J, Ahmad KA, et al: Math1 is essential for the development of hindbrain neurons critical for perinatal breathing. Neuron. 64:341–354. 2009. View Article : Google Scholar : PubMed/NCBI
38	Kong J, Crissey MA, Sepulveda AR and Lynch JP: Math1/Atoh1 contributes to intestinalization of esophageal keratinocytes by inducing the expression of Muc2 and Keratin-20. Dig Dis Sci. 57:845–857. 2012. View Article : Google Scholar :