Co-expression of ING4 and P53 enhances hypopharyngeal cancer chemosensitivity to cisplatin in vivo

Ren,Xin; Liu,Hao; Zhang,Mingjie; Wang,Mengjun; Ma,Shiyin

doi:10.3892/mmr.2016.5552

Co-expression of ING4 and P53 enhances hypopharyngeal cancer chemosensitivity to cisplatin in vivo

Authors:
- Xin Ren
- Hao Liu
- Mingjie Zhang
- Mengjun Wang
- Shiyin Ma
View Affiliations

Affiliations: Department of Otolaryngology, The First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui 233030, P.R. China, Faculty of Pharmacy, Bengbu Medical College, Bengbu, Anhui 233030, P.R. China
Published online on: July 27, 2016 https://doi.org/10.3892/mmr.2016.5552
Pages: 2431-2438
Copyright: © Ren et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Hypopharyngeal cancer is a distinct type of malignant head and neck tumor, which exhibits low sensitivity to anti-cancer drugs. The importance of developing methods for reducing chemotherapy resistance, and improving and enhancing prognosis has previously been emphasized and is considered a challenge for effective clinical treatment of hypopharyngeal cancer. The current study investigated the effects of co‑expression of inhibitor of growth protein 4 (ING4) and P53, a tumor suppressor gene, on chemosensitivity to cisplatin in human hypopharyngeal cancer xenografts in vivo, and the potential molecular mechanisms involved. A tumor model was established by injecting athymic nude mice with FADU human hypopharyngeal cancer cells. Five days after intratumoral and peritumoral injections of an empty adenoviral vector (Ad), Ad‑ING4‑P53, cisplatin, or a combination of Ad‑ING4‑P53 and cisplatin (Ad‑ING4‑P53 + cisplatin) every other day for 5 days, the mice were euthanized and their tumors, livers, and kidneys were removed. The tumor weights were used to calculate the inhibition rate, and the expression levels of ING4 and P53 were detected by reverse transcription‑polymerase chain reaction. Additionally, apoptotic cells were detected using terminal deoxynucleotidyl transferase dUTP nick end labeling, and immunohistochemistry determined the levels ING4, P53, B‑cell lymphoma‑2 (Bcl‑2) and Bcl‑2 associated X protein (Bax) protein expression. The results demonstrated increased expression of ING4 and P53 in the Ad‑ING4‑P53 groups compared with PBS and Ad groups, indicating successful introduction of the genes into the tumor cells. Notably, the Ad‑ING4‑P53 + cisplatin group exhibited a higher inhibition rate compared with the four other groups. The results of immunohistochemistry analysis demonstrated that Bax expression was increased and Bcl‑2 was decreased in the Ad‑ING4‑P53 + cisplatin group. This suggested that the enhanced cisplatin chemosensitivity with Ad-ING4-P53 gene therapy in hypopharyngeal cancer xenografts may be associated with apoptosis induction through upregulation of Bax expression and downregulation of Bcl‑2. The results of the present study indicated that gene therapy combined with cisplatin treatment may be a promising treatment for human hypopharyngeal cancer.

View Figures

View References

1	Genden EM, Ferlito A, Bradley PJ, Rinaldo A and Scully C: Neck disease and distant metastases. Oral Oncol. 39:207–212. 2003. View Article : Google Scholar : PubMed/NCBI
2	Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015. View Article : Google Scholar : PubMed/NCBI
3	Keereweer S, Kerrebijn JD, Al-Mamgani A, Sewnaik A, Baatenburg de Jong RJ and van Meerten E: Chemoradiation for advanced hypopharyngeal carcinoma: A retrospective study on efficacy, morbidity and quality of life. Eur Arch Otorhinolaryngol. 269:939–946. 2012. View Article : Google Scholar :
4	Hanna NN, Mauceri HJ, Wayne JD, Hallahan DE, Kufe DW and Weichselbaum RR: Virally directed cytosine deaminase/5-fluo-rocytosine gene therapy enhances radiation response in human cancer xenografts. Cancer Res. 57:4205–4209. 1997.PubMed/NCBI
5	Thurfjell N, Coates PJ, Boldrup L, Lindgren B, Bäcklund B, Uusitalo T, Mahani D, Dabelsteen E, Dahlqvist A, Sjöström B, et al: Function and importance of p63 in normal oral mucosa and squamous cell carcinoma of the head and neck. Adv Otorhinolaryngol. 62:49–57. 2005.
6	Luo XR, Li JS, Niu Y and Miao L: Adenovirus-mediated double suicide gene selectively kills gastric cancer cells. Asian Pac J Cancer Prev. 13:781–784. 2012. View Article : Google Scholar : PubMed/NCBI
7	Shiseki M, Nagashima M, Pedeux RM, Kitahama-Shiseki M, Miura K, Okamura S, Onogi H, Higashimoto Y, Appella E, Yokota J and Harris CC: p29ING4 and p28ING5 bind to p53 and p300, and enhance p53 activity. Cancer Res. 63:2373–2378. 2003.PubMed/NCBI
8	Garkavtsev I, Kozin SV, Chernova O, Xu L, Winkler F, Brown E, Barnett GH and Jain RK: The candidate tumour suppressor protein ING4 regulates brain tumour growth and angiogenesis. Nature. 428:328–332. 2004. View Article : Google Scholar : PubMed/NCBI
9	Unoki M, Shen JC, Zheng ZM and Harris CC: Novel splice variants of ING4 and their possible roles in the regulation of cell growth and motility. J Biol Chem. 281:34677–34686. 2006. View Article : Google Scholar : PubMed/NCBI
10	Colla S, Tagliaferri S, Morandi F, Lunghi P, Donofrio G, Martorana D, Mancini C, Lazzaretti M, Mazzera L, Ravanetti L, et al: The new tumor-suppressor gene inhibitor of growth family member 4 (ING4) regulates the production of proangiogenic molecules by myeloma cells and suppresses hypoxia-inducible factor-1 alpha (HIF-1alpha) activity: Involvement in myeloma-induced angio-genesis. Blood. 110:4464–4475. 2007. View Article : Google Scholar : PubMed/NCBI
11	Shen JC, Unoki M, Ythier D, Duperray A, Varticovski L, Kumamoto K, Pedeux R and Harris CC: Inhibitor of growth 4 suppresses cell spreading and cell migration by interacting with a novel binding partner, liprin alpha1. Cancer Res. 67:2552–2558. 2007. View Article : Google Scholar : PubMed/NCBI
12	Li J, Martinka M and Li G: Role of ING4 in human melanoma cell migration, invasion and patient survival. Carcinogenesis. 29:1373–1379. 2008. View Article : Google Scholar : PubMed/NCBI
13	Cai L, Li X, Zheng S, Wang Y, Wang Y, Li H, Yang J and Sun J: Inhibitor of growth 4 is involved in melanomagenesis and induces growth suppression and apoptosis in melanoma cell line M14. Melanoma Res. 19:1–7. 2009. View Article : Google Scholar : PubMed/NCBI
14	Wei Q, He W, Lu Y, Yao J and Cao X: Effect of the tumor suppressor gene ING4 on the proliferation of MCF-7 human breast cancer cells. Oncol Lett. 4:438–442. 2012.PubMed/NCBI
15	Huang J, Yang J, Ling C, Zhao D, Xie Y and You Z: The mechanism of inhibition effect of adenovirus-mediated ING4 on human lung adenocarcinoma xenografts in nude mice. Zhongguo Fei Ai Za Zhi. 17:142–147. 2014.In Chinese. PubMed/NCBI
16	Xu M, Xie Y, Sheng W, Miao J and Yang J: Adenovirus-mediated ING4 gene transfer in osteosarcoma suppresses tumor growth via induction of apoptosis and inhibition of tumor angiogenesis. Technol Cancer Res Treat. 14:369–378. 2015. View Article : Google Scholar
17	Gunduz M, Nagatsuka H, Demircan K, Gunduz E, Cengiz B, Ouchida M, Tsujigiwa H, Yamachika E, Fukushima K, Beder L, et al: Frequent deletion and down-regulation of ING4, a candidate tumor suppressor gene at 12p13, in head and neck squamous cell carcinomas. Gene. 356:109–117. 2005. View Article : Google Scholar : PubMed/NCBI
18	Vijayaraman KP, Veluchamy M, Murugesan P, Shanmugiah KP and Kasi PD: p53 exon 4 (codon 72) polymorphism and exon 7 (codon 249) mutation in breast cancer patients in southern region (Madurai) of Tamil Nadu. Asian Pac J Cancer Prev. 13:511–516. 2012. View Article : Google Scholar : PubMed/NCBI
19	Levine AJ, Finlay CA and Hinds PW: P53 is a tumor suppressor gene. Cell. 116(Suppl 2): S67–S69, 1 p following S69. 2004. View Article : Google Scholar : PubMed/NCBI
20	Yu M, Chen W and Zhang J: p53 gene therapy for pulmonary metastasis tumor from hepatocellular carcinoma. Anticancer Drugs. 21:882–884. 2010. View Article : Google Scholar : PubMed/NCBI
21	Oshima Y, Sasaki Y, Negishi H, Idogawa M, Toyota M, Yamashita T, Wada T, Nagoya S, Kawaguchi S, Yamashita T and Tokino T: Antitumor effect of adenovirus-mediated p53 family gene transfer on osteosarcoma cell lines. Cancer Biol Ther. 6:1058–1066. 2007. View Article : Google Scholar : PubMed/NCBI
22	Russell M, Berardi P, Gong W and Riabowol K: Grow-ING, Age-ING and Die-ING: ING proteins link cancer, senescence and apoptosis. Exp Cell Res. 312:951–961. 2006. View Article : Google Scholar : PubMed/NCBI
23	Soliman MA and Riabowol K: After a decade of study-ING, a PHD for a versatile family of proteins. Trends Biochem Sci. 32:509–519. 2007. View Article : Google Scholar : PubMed/NCBI
24	Sozzi G, Miozzo M, Donghi R, Pilotti S, Cariani CT, Pastorino U, Della Porta G and Pierotti MA: Deletions of 17p and p53 mutations in preneoplastic lesions of the lung. Cancer Res. 52:6079–6082. 1992.PubMed/NCBI
25	Jin ZJ: Addition in drug combination (author's transl). Zhongguo Yao Li Xue Bao. 1:70–76. 1980.In Chinese. PubMed/NCBI
26	Yin W, Zhang J, Jiang Y and Juan S: Combination therapy with low molecular weight heparin and Adriamycin results in decreased breast cancer cell metastasis in CH mice. Exp Ther Med. 8:1213–1218. 2014.PubMed/NCBI
27	Chan DA, Sutphin PD, Nguyen P, Turcotte S, Lai EW, Banh A, Reynolds GE, Chi JT, Wu J, Solow-Cordero DE, et al: Targeting GLUT1 and the Warburg effect in renal cell carcinoma by chemical synthetic lethality. Sci Transl Med. 3:94ra702011. View Article : Google Scholar : PubMed/NCBI
28	Shimanishi M, Ogi K, Sogabe Y, Kaneko T, Dehari H, Miyazaki A and Hiratsuka H: Silencing of GLUT-1 inhibits sensitization of oral cancer cells to cisplatin during hypoxia. J Oral Pathol Med. 42:382–388. 2013. View Article : Google Scholar
29	Wang YD, Li SJ and Liao JX: Inhibition of glucose transporter 1 (GLUT1) chemosensitized head and neck cancer cells to cisplatin. Technol Cancer Res Treat. 12:525–535. 2013.PubMed/NCBI
30	Zhang X, Xu LS, Wang ZQ, Wang KS, Li N, Cheng ZH, Huang SZ, Wei DZ and Han ZG: ING4 induces G2/M cell cycle arrest and enhances the chemosensitivity to DNA-damage agents in HepG2 cells. FEBS Lett. 570:7–12. 2004. View Article : Google Scholar : PubMed/NCBI
31	Zhou X, Gu Y and Zhang SL: Association between p53 codon 72 polymorphism and cervical cancer risk among Asians: A HuGE review and meta-analysis. Asian Pac J Cancer Prev. 13:4909–4914. 2012. View Article : Google Scholar : PubMed/NCBI
32	Kraljević Pavelić S, Marjanović M, Poznić M and Kralj M: Adeno-virally mediated p53 overexpression diversely influence the cell cycle of HEp-2 and CAL 27 cell lines upon cisplatin and methotrexate treatment. J Cancer Res Clin Oncol. 135:1747–1761. 2009. View Article : Google Scholar
33	Rudin CM and Thompson CB: Apoptosis and disease: Regulation and clinical relevance of programmed cell death. Ann Rev Med. 48:267–281. 1997. View Article : Google Scholar : PubMed/NCBI
34	Tse C, Shoemaker AR, Adickes J, Anderson MG, Chen J, Jin S, Johnson EF, Marsh KC, Mitten MJ, Nimmer P, et al: ABT-263: A potent and orally bioavailable Bcl-2 family inhibitor. Cancer Res. 68:3421–3428. 2008. View Article : Google Scholar : PubMed/NCBI
35	Cosulich SC, Savory PJ and Clarke PR: Bcl-2 regulates amplification of caspase activation by cytochrome c. Curr Biol. 9:147–150. 1999. View Article : Google Scholar : PubMed/NCBI
36	Lozier JN, Csako G, Mondoro TH, Krizek DM, Metzger ME, Costello R, Vostal JG, Rick ME, Donahue RE and Morgan RA: Toxicity of a first-generation adenoviral vector in rhesus macaques. Hum Gene Ther. 13:113–124. 2002. View Article : Google Scholar : PubMed/NCBI
37	Li Y, Shao JY, Liu RY, Zhou L, Chai LP, Li HL, Han HY, Huang BJ, Zeng MS, Zhu XF, et al: Evaluation of long-term toxicity of Ad/hIFN-, an Adenoviral vector encoding the human interferon-gamma gene, in nonhuman primates. Hum Gene Ther. 19:827–839. 2008. View Article : Google Scholar : PubMed/NCBI
38	Haisma HJ, Boesjes M, Beerens AM, van der Strate BW, Curiel DT, Plüddemann A, Gordon S and Bellu AR: Scavenger receptor A: A new route for adenovirus 5. Mol Pharm. 6:366–374. 2009. View Article : Google Scholar : PubMed/NCBI
39	Lieber A, He CY, Meuse L, Schowalter D, Kirillova I, Winther B and Kay MA: The role of Kupffer cell activation and viral gene expression in early liver toxicity after infusion of recombinant adenovirus vectors. J Virol. 71:8798–8807. 1997.PubMed/NCBI
40	Zhu Y, Lv H, Xie Y, Sheng W, Xiang J and Yang J: Enhanced tumor suppression by an ING4/IL-24 bicistronic adenovirus-mediated gene cotransfer in human non-small cell lung cancer cells. Cancer Gene Ther. 18:627–636. 2011. View Article : Google Scholar : PubMed/NCBI