Slug inhibition increases radiosensitivity of oral squamous cell carcinoma cells by upregulating PUMA

Jiang,Fangfang; Zhou,Lijie; Wei,Changbo; Zhao,Wei; Yu,Dongsheng

doi:10.3892/ijo.2016.3570

Slug inhibition increases radiosensitivity of oral squamous cell carcinoma cells by upregulating PUMA

Authors:
- Fangfang Jiang
- Lijie Zhou
- Changbo Wei
- Wei Zhao
- Dongsheng Yu
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Affiliations: Department of Oral and Maxillofacial Surgery, Guanghua School of Stomatology, Hospital of Stomatology, Guangdong Provincial Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, Guangdong 510055, P.R. China
Published online on: June 8, 2016 https://doi.org/10.3892/ijo.2016.3570
Pages: 709-719

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Abstract

As a new strategy, radio-gene therapy was widely used for the treatment of cancer patients in recent few years. Slug was involved in the radioresistance of various cancers and has been found to have an anti-apoptotic effect. This study aims to investigate whether the modulation of Slug expression by siRNA affects oral squamous cell carcinoma sensitivity to X-ray irradiation through upregulating PUMA. Two oral squamous cell carcinoma cell lines (HSC3 and HSC6) were transfected with small interfering RNA (siRNA) targeting Slug and subjected to radiotherapy in vitro. After transfection with Slug siRNA, both HSC3 and HSC6 cells showed relatively lower expression of Slug and higher expression of PUMA. The Slug siRNA transfected cells showed decreased survival and proliferation rates, an increased apoptosis rate and enhanced radiosensitivity to X-ray irradiation. Our results revealed that Slug siRNA transfection in combination with radiation increased the expression of PUMA, which contributed to radiosensitivity of oral squamous cell carcinoma cells. Thus, controlling the expression of Slug might contribute to enhance sensitivity of HSC3 and HSC6 cells toward X-ray irradiation in vitro by upregulating PUMA.

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1	Zhao Y, Li Z, Sheng W, Miao J and Yang J: Radiosensitivity by ING4-IL-24 bicistronic adenovirus-mediated gene cotransfer on human breast cancer cells. Cancer Gene Ther. 20:38–45. 2013. View Article : Google Scholar
2	Wu J, Zhang JY, Yin L, Wu JZ, Guo WJ, Wu JF, Chen M, Xia YY, Tang JH, Ma YC, et al: HAP1 gene expression is associated with radiosensitivity in breast cancer cells. Biochem Biophys Res Commun. 456:162–166. 2015. View Article : Google Scholar
3	Finnon P, Kabacik S, MacKay A, Raffy C, A'Hern R, Owen R, Badie C, Yarnold J and Bouffler S: Correlation of in vitro lymphocyte radiosensitivity and gene expression with late normal tissue reactions following curative radiotherapy for breast cancer. Radiother Oncol. 105:329–336. 2012. View Article : Google Scholar : PubMed/NCBI
4	Ganapathy S, Xiao S, Yang M, Qi M, Choi DE, Ha CS, Little JB and Yuan ZM: A low-dose arsenic-induced p53 protein-mediated metabolic mechanism of radiotherapy protection. J Biol Chem. 289:5340–5347. 2014. View Article : Google Scholar : PubMed/NCBI
5	Agra IM, Filho JG, Martins EP and Kowalski LP: Second salvage surgery for re-recurrent oral cavity and oropharynx carcinoma. Head Neck. 32:997–1002. 2010. View Article : Google Scholar : PubMed/NCBI
6	Brun SN, Markant SL, Esparza LA, Garcia G, Terry D, Huang JM, Pavlyukov MS, Li XN, Grant GA, Crawford JR, et al: Survivin as a therapeutic target in Sonic hedgehog-driven medulloblastoma. Oncogene. 34:3770–3779. 2015. View Article : Google Scholar :
7	Markant SL, Esparza LA, Sun J, Barton KL, McCoig LM, Grant GA, Crawford JR, Levy ML, Northcott PA, Shih D, et al: Targeting sonic hedgehog-associated medulloblastoma through inhibition of Aurora and Polo-like kinases. Cancer Res. 73:6310–6322. 2013. View Article : Google Scholar : PubMed/NCBI
8	Xu T, Fan B, Lv C and Xiao D: Slug mediates nasopharyngeal carcinoma radioresistance via downregulation of PUMA in a p53-dependent and -independent manner. Oncol Rep. 33:2631–2638. 2015.PubMed/NCBI
9	Lee SH, Kim DY, Jing F, Kim H, Yun CO, Han DJ and Choi EY: Del-1 overexpression potentiates lung cancer cell proliferation and invasion. Biochem Biophys Res Commun. 468:92–98. 2015. View Article : Google Scholar : PubMed/NCBI
10	Chiang C and Ayyanathan K: Characterization of the E-box binding affinity to snag-zinc finger proteins. Mol Biol (Mosk). 46:907–914. 2012. View Article : Google Scholar
11	Pérez-Losada J, Sánchez-Martín M, Pérez-Caro M, Pérez-Mancera PA and Sánchez-García I: The radioresistance biological function of the SCF/kit signaling pathway is mediated by the zinc-finger transcription factor Slug. Oncogene. 22:4205–4211. 2003. View Article : Google Scholar : PubMed/NCBI
12	Pérez-Caro M, Bermejo-Rodríguez C, González-Herrero I, Sánchez-Beato M, Piris MA and Sánchez-García I: Transcriptomal profiling of the cellular response to DNA damage mediated by Slug (Snai2). Br J Cancer. 98:480–488. 2008. View Article : Google Scholar : PubMed/NCBI
13	Yu J, Zhang L, Hwang PM, Kinzler KW and Vogelstein B: PUMA induces the rapid apoptosis of colorectal cancer cells. Mol Cell. 7:673–682. 2001. View Article : Google Scholar : PubMed/NCBI
14	Feng J, Meng C and Xing D: Aβ induces PUMA activation: A new mechanism for Aβ-mediated neuronal apoptosis. Neurobiol Aging. 36:789–800. 2015. View Article : Google Scholar
15	Zhang K, Zhang B, Lu Y, Sun C, Zhao W, Jiao X, Hu J, Mu P, Lu H and Zhou C: Slug inhibition upregulates radiation-induced PUMA activity leading to apoptosis in cholangiocarcinomas. Med Oncol. 28(Suppl 1): S301–S309. 2011. View Article : Google Scholar
16	Kuribayashi K, Finnberg N, Jeffers JR, Zambetti GP and El-Deiry WS: The relative contribution of pro-apoptotic p53-target genes in the triggering of apoptosis following DNA damage in vitro and in vivo. Cell Cycle. 10:2380–2389. 2011. View Article : Google Scholar : PubMed/NCBI
17	Gauger KJ and Schneider SS: Tumour supressor secreted frizzled related protein 1 regulates p53-mediated apoptosis. Cell Biol Int. 38:124–130. 2014. View Article : Google Scholar
18	Wang R, Wang X, Li B, Lin F, Dong K, Gao P and Zhang HZ: Tumor-specific adenovirus-mediated PUMA gene transfer using the survivin promoter enhances radiosensitivity of breast cancer cells in vitro and in vivo. Breast Cancer Res Treat. 117:45–54. 2009. View Article : Google Scholar
19	Yu J, Yue W, Wu B and Zhang L: PUMA sensitizes lung cancer cells to chemotherapeutic agents and irradiation. Clin Cancer Res. 12:2928–2936. 2006. View Article : Google Scholar : PubMed/NCBI
20	Wu WS, Heinrichs S, Xu D, Garrison SP, Zambetti GP, Adams JM and Look AT: Slug antagonizes p53-mediated apoptosis of hematopoietic progenitors by repressing PUMA. Cell. 123:641–653. 2005. View Article : Google Scholar : PubMed/NCBI
21	Arienti C, Tesei A, Carloni S, Ulivi P, Romeo A, Ghigi G, Menghi E, Sarnelli A, Parisi E, Silvestrini R, et al: SLUG silencing increases radiosensitivity of melanoma cells in vitro. Cell Oncol (Dordr). 36:131–139. 2013. View Article : Google Scholar
22	Dillon JK, Brown CB, McDonald TM, Ludwig DC, Clark PJ, Leroux BG and Futran ND: How does the close surgical margin impact recurrence and survival when treating oral squamous cell carcinoma? J Oral Maxillofac Surg. 73:1182–1188. 2015. View Article : Google Scholar : PubMed/NCBI
23	Inhestern J, Oertel K, Stemmann V, Schmalenberg H, Dietz A, Rotter N, Veit J, Görner M, Sudhoff H, Junghanss C, et al: Prognostic role of circulating tumor cells during induction chemotherapy followed by curative surgery combined with postoperative radiotherapy in patients with locally advanced oral and oropharyngeal squamous cell cancer. PLoS One. 10:e01329012015. View Article : Google Scholar : PubMed/NCBI
24	Huang SH and O'Sullivan B: Oral cancer: Current role of radiotherapy and chemotherapy. Med Oral Patol Oral Cir Bucal. 18:e233–e240. 2013. View Article : Google Scholar : PubMed/NCBI
25	Lo WL, Kao SY, Chi LY, Wong YK and Chang RC: Outcomes of oral squamous cell carcinoma in Taiwan after surgical therapy: Factors affecting survival. J Oral Maxillofac Surg. 61:751–758. 2003. View Article : Google Scholar : PubMed/NCBI
26	Zhang P, Liu H, Xia F, Zhang QW, Zhang YY, Zhao Q, Chao ZH, Jiang ZW and Jiang CC: Epithelial-mesenchymal transition is necessary for acquired resistance to cisplatin and increases the metastatic potential of nasopharyngeal carcinoma cells. Int J Mol Med. 33:151–159. 2014.
27	Kurrey NK, Jalgaonkar SP, Joglekar AV, Ghanate AD, Chaskar PD, Doiphode RY and Bapat SA: Snail and slug mediate radioresistance and chemoresistance by antagonizing p53-mediated apoptosis and acquiring a stem-like phenotype in ovarian cancer cells. Stem Cells. 27:2059–2068. 2009. View Article : Google Scholar : PubMed/NCBI
28	Kim S, Yao J, Suyama K, Qian X, Qian BZ, Bandyopadhyay S, Loudig O, De Leon-Rodriguez C, Zhou ZN, Segall J, et al: Slug promotes survival during metastasis through suppression of Puma-mediated apoptosis. Cancer Res. 74:3695–3706. 2014. View Article : Google Scholar : PubMed/NCBI
29	Yu J and Zhang L: No PUMA, no death: Implications for p53-dependent apoptosis. Cancer Cell. 4:248–249. 2003. View Article : Google Scholar : PubMed/NCBI
30	Akhter R, Sanphui P and Biswas SC: The essential role of p53-up-regulated modulator of apoptosis (Puma) and its regulation by FoxO3a transcription factor in β-amyloid-induced neuron death. J Biol Chem. 289:10812–10822. 2014. View Article : Google Scholar : PubMed/NCBI
31	Nakano K and Vousden KH: PUMA, a novel proapoptotic gene, is induced by p53. Mol Cell. 7:683–694. 2001. View Article : Google Scholar : PubMed/NCBI
32	Yu J, Wang Z, Kinzler KW, Vogelstein B and Zhang L: PUMA mediates the apoptotic response to p53 in colorectal cancer cells. Proc Natl Acad Sci USA. 100:1931–1936. 2003. View Article : Google Scholar : PubMed/NCBI
33	Chen L, Willis SN, Wei A, Smith BJ, Fletcher JI, Hinds MG, Colman PM, Day CL, Adams JM and Huang DC: Differential targeting of prosurvival Bcl-2 proteins by their BH3-only ligands allows complementary apoptotic function. Mol Cell. 17:393–403. 2005. View Article : Google Scholar : PubMed/NCBI
34	Funamizu N, Lacy CR, Kamada M, Yanaga K and Manome Y: MicroRNA-203 induces apoptosis by upregulating Puma expression in colon and lung cancer cells. Int J Oncol. 47:1981–1988. 2015.PubMed/NCBI