Photothermal therapy with AuNRs and EGFRmAb-AuNRs inhibits subcutaneous transplantable hypopharyngeal tumors in nude mice

Zhang,Yingying; He,Jiayan; Wang,Yu; Wen,Jinya; Zou,Yingying; Yang,Zhihong; He,Xiaoguang

doi:10.3892/ijo.2018.4559

Photothermal therapy with AuNRs and EGFRmAb-AuNRs inhibits subcutaneous transplantable hypopharyngeal tumors in nude mice

Authors:
- Yingying Zhang
- Jiayan He
- Yu Wang
- Jinya Wen
- Yingying Zou
- Zhihong Yang
- Xiaoguang He
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Affiliations: Department of Pathology and Pathophysiology, School of Basic Medical Science, Kunming Medical University, Kunming, Yunnan 650500, P.R. China, Department of Head and Neck Surgery, The First Affiliated Hospital of Kunming Medical University, Kunming, Yunnan 650032, P.R. China
Published online on: September 12, 2018 https://doi.org/10.3892/ijo.2018.4559
Pages: 2647-2658

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Abstract

The present study aimed to investigate the effects of photothermal therapy with gold nanorods (AuNRs) or epidermal growth factor receptor monoclonal antibody‑conjugated AuNRs (EGFRmAb‑AuNRs) on hypopharyngeal carcinoma (HC) in nude mice. In addition, the associated signaling pathways were explored. Briefly, a subcutaneous transplantable hypopharyngeal tumor model was established in nude mice injected with FaDu human HC cells. A total of 70 nude mice were randomly divided into seven groups, each of which received a different treatment. Mice were treated with AuNRs, locally or through intravenous injection, whereas EGFRmAb or EGFRmAb‑AuNRs were only administered locally. Near infrared spectroscopy (NIR) was also applied for plasmonic photothermal therapy (PPTT). The growth curve and the inhibitory rate for tumor growth were used to evaluate the effects of each treatment. Flow cytometry and the terminal‑deoxynucleotidyl transferase dUTP nick end labeling assay were adopted to detect apoptosis of cells in the transplanted tumors. Reverse transcription‑quantitative polymerase chain reaction and western blotting were used to determine the mRNA and protein expression levels of target genes, respectively. Local treatment with AuNRs + NIR or EGFRmAb significantly inhibited tumor growth, and EGFRmAb conjugation further increased the inhibitory effects. Furthermore, there was a significant increase in apoptosis of tumor cells in the AuNRs + NIR, EGFRmAb and EGFRmAb‑AuNRs + NIR groups; treatment with EGFRmAb‑AuNRs + NIR induced the highest apoptotic effect. Mechanistic studies indicated that EGFRmAb‑AuNRs + NIR may inhibit tumors through the AKT serine/threonine kinase (Akt) and DNA damage signaling pathways. In the AKT pathway, the mRNA and protein expression levels of phosphatase and tensin homolog were increased, whereas the expression levels of Akt and glycogen synthase kinase 3β were decreased. In the DNA damage signaling pathway, the mRNA and protein expression levels of ATR serine/threonine kinase, checkpoint kinase 1 and p53 were enhanced, whereas phosphorylated‑p53 protein expression was reduced. The present findings indicated that AuNRs + NIR inhibited HC tumor growth, and conjugating EGFRmAb to AuNRs further enhanced the inhibitory effects. EGFRmAb conjugation may increase the antitumor effects of AuNRs‑induced PPTT by downregulating the phosphatidylinositol‑3‑kinase/Akt pathway and upregulating the DNA damage pathway. These findings may provide novel insights into tumor‑targeting PPTT in vivo.

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1	Rivière D, Mancini J, Santini L, Giovanni A, Dessi P and Fakhry N: Lymph-node metastasis following total laryngectomy and total pharyngolaryngectomy for laryngeal and hypopharyngeal squamous cell carcinoma: Frequency, distribution and risk factors. Eur Ann Otorhinolaryngol Head Neck Dis. 135:163–166. 2018. View Article : Google Scholar
2	Krstevska V, Stojkovski I and Lukarski D: Concurrent radiochemotherapy in advanced hypopharyngeal cancer. Radiat Oncol. 5:392010. View Article : Google Scholar : PubMed/NCBI
3	Cabral RM and Baptista PV: Anti-cancer precision theranostics: A focus on multifunctional gold nanoparticles. Expert Rev Mol Diagn. 14:1041–1052. 2014. View Article : Google Scholar : PubMed/NCBI
4	Aioub M and El-Sayed MA: A real-time surface enhanced raman spectroscopy study of plasmonic photothermal cell death using targeted gold nanoparticles. J Am Chem Soc. 138:1258–1264. 2016. View Article : Google Scholar : PubMed/NCBI
5	Brand TM, Iida M, Li C and Wheeler DL: The nuclear epidermal growth factor receptor signaling network and its role in cancer. Discov Med. 12:419–432. 2011.PubMed/NCBI
6	Zhang S, Li Y, He X, Dong S, Huang Y, Li X, Li Y, Jin C, Zhang Y and Wang Y: Photothermolysis mediated by gold nanorods modified with EGFR monoclonal antibody induces Hep-2 cells apoptosis in vitro and in vivo. Int J Nanomedicine. 9:1931–1946. 2014.PubMed/NCBI
7	Normanno N, De Luca A, Bianco C, Strizzi L, Mancino M, Maiello MR, Carotenuto A, De Feo G, Caponigro F and Salomon DS: Epidermal growth factor receptor (EGFR) signaling in cancer. Gene. 366:2–16. 2006. View Article : Google Scholar
8	Garrett TP, McKern NM, Lou M, Elleman TC, Adams TE, Lovrecz GO, Zhu HJ, Walker F, Frenkel MJ, Hoyne PA, et al: Crystal structure of a truncated epidermal growth factor receptor extracellular domain bound to transforming growth factor alpha. Cell. 110:763–773. 2002. View Article : Google Scholar : PubMed/NCBI
9	Sourbier C, Lindner V, Lang H, Agouni A, Schordan E, Danilin S, Rothhut S, Jacqmin D, Helwig JJ and Massfelder T: The phosphoinositide 3-kinase/Akt pathway: A new target in human renal cell carcinoma therapy. Cancer Res. 66:5130–5142. 2006. View Article : Google Scholar : PubMed/NCBI
10	Hotfilder M, Sondermann P, Senss A, van Valen F, Jürgens H and Vormoor J: PI3K/AKT is involved in mediating survival signals that rescue Ewing tumour cells from fibroblast growth factor 2-induced cell death. Br J Cancer. 92:705–710. 2005. View Article : Google Scholar : PubMed/NCBI
11	Sandra CM, Eduardo CC, Simon HO, Teresa RA, Antonio NC, Lijanova IV and Marcos MG: Anticancer activity and anti-inflammatory studies of 5‑aryl‑1,4‑benzodiazepine derivatives. Anticancer Agents Med Chem. 12:611–618. 2012. View Article : Google Scholar : PubMed/NCBI
12	Clemens MJ: Targets and mechanisms for the regulation of translation in malignant transformation. Oncogene. 23:3180–3188. 2004. View Article : Google Scholar : PubMed/NCBI
13	Vignard J, Mirey G and Salles B: Ionizing-radiation induced DNA double-strand breaks: A direct and indirect lighting up. Radiother Oncol. 108:362–369. 2013. View Article : Google Scholar : PubMed/NCBI
14	Ciccia A and Elledge SJ: The DNA damage response: Making it safe to play with knives. Mol Cell. 40:179–204. 2010. View Article : Google Scholar : PubMed/NCBI
15	Abraham RT: Cell cycle checkpoint signaling through the ATM and ATR kinases. Genes Dev. 15:2177–2196. 2001. View Article : Google Scholar : PubMed/NCBI
16	Demoulin B, Hermant M, Castrogiovanni C, Staudt C and Dumont P: Resveratrol induces DNA damage in colon cancer cells by poisoning topoisomerase II and activates the ATM kinase to trigger p53-dependent apoptosis. Toxicol In Vitro. 29:1156–1165. 2015. View Article : Google Scholar : PubMed/NCBI
17	Zhang Y, Cong L, He J, Wang Y, Zou Y, Yang Z, Hu Y, Zhang S and He X: Photothermal treatment with EGFRmAb-AuNPs induces apoptosis in hypopharyngeal carcinoma cells via PI3K/AKT/mTOR and DNA damage response pathways. Acta Biochim Biophys Sin (Shanghai). 50:567–578. 2018. View Article : Google Scholar
18	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar
19	Durr NJ, Larson T, Smith DK, Korgel BA, Sokolov K and Ben-Yakar A: Two-photon luminescence imaging of cancer cells using molecularly targeted gold nanorods. Nano Lett. 7:941–945. 2007. View Article : Google Scholar : PubMed/NCBI
20	Dickerson EB, Dreaden EC, Huang X, El-Sayed IH, Chu H, Pushpanketh S, McDonald JF and El-Sayed MA: Gold nanorod assisted near-infrared plasmonic photothermal therapy (PPTT) of squamous cell carcinoma in mice. Cancer Lett. 269:57–66. 2008. View Article : Google Scholar : PubMed/NCBI
21	Castellani R, Visscher DW, Wykes S, Sarkar FH and Crissman JD: Interaction of transforming growth factor-alpha and epidermal growth factor receptor in breast carcinoma. An immunohistologic study. Cancer. 73:344–349. 1994. View Article : Google Scholar : PubMed/NCBI
22	Giampieri R, Scartozzi M, Del Prete M, Maccaroni E, Bittoni A, Faloppi L, Bianconi M, Cecchini L and Cascinu S: Molecular biomarkers of resistance to anti-EGFR treatment in metastatic colorectal cancer, from classical to innovation. Crit Rev Oncol Hematol. 88:272–283. 2013. View Article : Google Scholar : PubMed/NCBI
23	Bonner JA, Harari PM, Giralt J, Azarnia N, Shin DM, Cohen RB, Jones CU, Sur R, Raben D, Jassem J, et al: Radiotherapy plus cetuximab for squamous-cell carcinoma of the head and neck. N Engl J Med. 354:567–578. 2006. View Article : Google Scholar : PubMed/NCBI
24	Bonner JA, Harari PM, Giralt J, Cohen RB, Jones CU, Sur RK, Raben D, Baselga J, Spencer SA, Zhu J, et al: Radiotherapy plus cetuximab for locoregionally advanced head and neck cancer: 5-year survival data from a phase 3 randomised trial, and relation between cetuximab-induced rash and survival. Lancet Oncol. 11:21–28. 2010. View Article : Google Scholar
25	Ahlmann E, Patzakis M, Roidis N, Shepherd L and Holtom P: Comparison of anterior and posterior iliac crest bone grafts in terms of harvest-site morbidity and functional outcomes. J Bone Joint Surg Am. 84-A:716–720. 2002. View Article : Google Scholar : PubMed/NCBI
26	LoPiccolo J, Blumenthal GM, Bernstein WB and Dennis PA: Targeting the PI3K/Akt/mTOR pathway: effective combinations and clinical considerations. Drug Resist Updat. 11:32–50. 2008. View Article : Google Scholar : PubMed/NCBI
27	Catasus L, D’Angelo E, Pons C, Espinosa I and Prat J: Expression profiling of 22 genes involved in the PI3K‑AKT pathway identifies two subgroups of high‑grade endometrial carcinomas with different molecular alterations. Mod Pathol. 23:694–702. 2010. View Article : Google Scholar : PubMed/NCBI
28	Priulla M, Calastretti A, Bruno P, Azzariti A, Paradiso A, Canti G and Nicolin A: Preferential chemosensitization of PTEN-mutated prostate cells by silencing the Akt kinase. Prostate. 67:782–789. 2007. View Article : Google Scholar : PubMed/NCBI
29	Zhang D and Fan D: Multidrug resistance in gastric cancer: Recent research advances and ongoing therapeutic challenges. Expert Rev Anticancer Ther. 7:1369–1378. 2007. View Article : Google Scholar : PubMed/NCBI
30	Yang L, Dan HC, Sun M, Liu Q, Sun XM, Feldman RI, Hamilton AD, Polokoff M, Nicosia SV, Herlyn M, et al: Akt/protein kinase B signaling inhibitor-2, a selective small molecule inhibitor of Akt signaling with antitumor activity in cancer cells overexpressing Akt. Cancer Res. 64:4394–4399. 2004. View Article : Google Scholar : PubMed/NCBI
31	Liu X, Dai X and Wu B: Study of 5-Aza-CdR on transcription regulation of RASSF1A gene in the BIU87 cell line. Urol Int. 82:108–112. 2009. View Article : Google Scholar : PubMed/NCBI
32	Liao W, McNutt MA and Zhu WG: The comet assay: A sensitive method for detecting DNA damage in individual cells. Methods. 48:46–53. 2009. View Article : Google Scholar : PubMed/NCBI
33	Lieberman HB: DNA damage repair and response proteins as targets for cancer therapy. Curr Med Chem. 15:360–367. 2008. View Article : Google Scholar : PubMed/NCBI
34	Matsumoto K, Nagahara T, Okano J and Murawaki Y: The growth inhibition of hepatocellular and cholangiocellular carcinoma cells by gemcitabine and the roles of extracellular signal-regulated and checkpoint kinases. Oncol Rep. 20:863–872. 2008.PubMed/NCBI
35	Aioub M, Panikkanvalappil SR and El-Sayed MA: Platinum-coated gold nanorods: Efficient reactive oxygen scavengers that prevent oxidative damage toward healthy, untreated cells during plasmonic photothermal therapy. ACS Nano. 11:579–586. 2017. View Article : Google Scholar
36	Schieber M and Chandel NS: ROS function in redox signaling and oxidative stress. Curr Biol. 24:R453–R462. 2014. View Article : Google Scholar : PubMed/NCBI