Standardization of A375 human melanoma models on chicken embryo chorioallantoic membrane and Balb/c nude mice

Avram,Stefana; Coricovac,Dorina‑Elena; Pavel,Ioana  Zinuca; Pinzaru,Iulia; Ghiulai,Roxana; Baderca,Flavia; Soica,Codruta; Muntean,Danina; Branisteanu,Daciana  E.; Spandidos,Demetrios  A.; Tsatsakis,Aristides M.; Dehelean,Cristina  Adriana

doi:10.3892/or.2017.5658

Standardization of A375 human melanoma models on chicken embryo chorioallantoic membrane and Balb/c nude mice

Authors:
- Stefana Avram
- Dorina‑Elena Coricovac
- Ioana Zinuca Pavel
- Iulia Pinzaru
- Roxana Ghiulai
- Flavia Baderca
- Codruta Soica
- Danina Muntean
- Daciana E. Branisteanu
- Demetrios A. Spandidos
- Aristides M. Tsatsakis
- Cristina Adriana Dehelean
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Affiliations: Department of Pharmacognosy, Faculty of Pharmacy, ‘Victor Babeș’ University of Medicine and Pharmacy Timisoara, Timisoara 30004, Romania, Department of Toxicology, Faculty of Pharmacy, ‘Victor Babeș’ University of Medicine and Pharmacy Timisoara, Timisoara 30004, Romania, Department of Pharmaceutical Chemistry, Faculty of Pharmacy, ‘Victor Babeș’ University of Medicine and Pharmacy Timisoara, Timisoara 30004, Romania, Department of Microscopic Morphology, Faculty of Medicine, ‘Victor Babeș’ University of Medicine and Pharmacy Timisoara, Timisoara 300041, Romania, Department of Pathophysiology, Faculty of Medicine, ‘Victor Babeș’ University of Medicine and Pharmacy Timisoara, Timisoara 300041, Romania, Department of Dermatology and Venereology, ‘Grigore T. Popa’ University of Medicine and Pharmacy Iasi, Iasi 7000115, Romania, Laboratory of Clinical Virology, School of Medicine, University of Crete, Heraklion 71003, Greece, Department of Forensic Sciences and Toxicology, Faculty of Medicine, University of Crete, Heraklion 71003, Greece
Published online on: May 22, 2017 https://doi.org/10.3892/or.2017.5658
Pages: 89-99
Copyright: © Avram et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Cutaneous melanoma is a metastatic disease characterized by high resistance to treatment, the incidence of which has alarmingly increased worldwide over the past years. A thorough characterization of tumor onset, progression and metastasis is compulsory to overcome the gaps existent in melanoma biology. The present study suggests a well‑established protocol and a detailed histological description of human melanoma models in ovo and in vivo obtained by the inoculation of A375 cells to chick embryo chorioallantoic membrane (CAM) and Balb/c nude mice. The inoculation of A375 cells on CAM led to the formation of compact primary and secondary tumors on day 4 post‑inoculation, with mean surface area values of 2.2±0.4 mm2 and 1.5±0.3 mm2, respectively. Moreover, the vessels around the tumors presented a spike wheel pattern, indicating a strong angiogenic reaction. All the injected mice, apart from one, developed solid polypoid primary tumors with lobulated surfaces and intense vascularization, and achromic epithelioid malignant melanocytes with vesiculous nuclei and necrosis area were detected. Metastasis was histologically confirmed in only 30% of the mice with the tumor xenografts. These data indicate that the standardization protocols proposed are complex and reproducible, and can be further employed for the therapeutic surveillance of antiangiogenic and anticancer agents.

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1	Baderca F, Cojocaru S, Lazăr E, Lăzureanu C, Lighezan R, Alexa A, Raica M and Nicola T: Amelanotic vulvar melanoma: Case report and review of the literature. Rom J Morphol Embryol. 49:219–228. 2008.PubMed/NCBI
2	Baderca F, Vincze D, Balica N and Solovan C: Mucosal melanomas in the elderly: Challenging cases and review of the literature. Clin Interv Aging. 9:929–937. 2014. View Article : Google Scholar : PubMed/NCBI
3	Kuzu OF, Nguyen FD, Noory MA and Sharma A: Current State of Animal (Mouse) Modeling in Melanoma Research. Cancer Growth Metastasis. 8 Suppl 1:81–94. 2015.PubMed/NCBI
4	Potrony M, Badenas C, Aguilera P, Puig-Butille JA, Carrera C, Malvehy J and Puig S: Update in genetic susceptibility in melanoma. Ann Transl Med. 3:2102015.PubMed/NCBI
5	Delyon J, Varna M, Feugeas JP, Sadoux A, Yahiaoui S, Podgorniak MP, Leclert G, Dorval SM, Dumaz N, Janin A, et al: Validation of a preclinical model for assessment of drug efficacy in melanoma. Oncotarget. 7:13069–13081. 2016.PubMed/NCBI
6	Candido S, Rapisarda V, Marconi A, Malaponte G, Bevelacqua V, Gangemi P, Scalisi A, McCubrey JA, Maestro R, Spandidos DA, et al: Analysis of the B-RafV600E mutation in cutaneous melanoma patients with occupational sun exposure. Oncol Rep. 31:1079–1082. 2014.PubMed/NCBI
7	Palmieri G, Colombino M, Sini MC, Ascierto PA, Lissia A and Cossu A: Targeted Therapies in Melanoma: Successes and PitfallsMelanoma-From Early Detection to Treatment. Duc GHT: InTech Open, Reykjavik. pp. 29–58. 2013
8	Sun C, Wang L, Huang S, Heynen GJ, Prahallad A, Robert C, Haanen J, Blank C, Wesseling J, Willems SM, et al: Reversible and adaptive resistance to BRAF(V600E) inhibition in melanoma. Nature. 508:118–122. 2014. View Article : Google Scholar : PubMed/NCBI
9	Russo A, Ficili B, Candido S, Pezzino FM, Guarneri C, Biondi A, Travali S, McCubrey JA, Spandidos DA and Libra M: Emerging targeted therapies for melanoma treatment (review). Int J Oncol. 45:516–524. 2014.PubMed/NCBI
10	Chalkiadaki G, Nikitovic D, Katonis P, Berdiaki A, Tsatsakis A, Kotsikogianni I, Karamanos NK and Tzanakakis GN: Low molecular weight heparin inhibits melanoma cell adhesion and migration through a PKCa/JNK signaling pathway inducing actin cytoskeleton changes. Cancer Lett. 312:235–244. 2011. View Article : Google Scholar : PubMed/NCBI
11	Yamanaka K, Nakahara T, Yamauchi T, Kita A, Takeuchi M, Kiyonaga F, Kaneko N and Sasamata M: Antitumor activity of YM155, a selective small-molecule survivin suppressant, alone and in combination with docetaxel in human malignant melanoma models. Clin Cancer Res. 17:5423–5431. 2011. View Article : Google Scholar : PubMed/NCBI
12	Ribatti D: The CAM assay in the study of angiogenesis and metastasis. The Chick Embryo Chorioallantoic Membrane in the Study of Angiogenesis and Metastasis. Springer. (Netherlands). 2010. View Article : Google Scholar
13	Murphy JB and Rous P: The behavior of chicken sarcoma implanted in the developing embryo. J Exp Med. 15:119–132. 1912. View Article : Google Scholar : PubMed/NCBI
14	Deryugina EI and Quigley JP: Chick embryo chorioallantoic membrane model systems to study and visualize human tumor cell metastasis. Histochem Cell Biol. 130:1119–1130. 2008. View Article : Google Scholar : PubMed/NCBI
15	Klingenberg M, Becker J, Eberth S, Kube D and Wilting J: The chick chorioallantoic membrane as an in vivo xenograft model for Burkitt lymphoma. BMC Cancer. 14:3392014. View Article : Google Scholar : PubMed/NCBI
16	Ribatti D, Nico B, Cimpean AM, Raica M, Crivellato E, Ruggieri S and Vacca A: B16-F10 melanoma cells contribute to the new formation of blood vessels in the chick embryo chorioallantoic membrane through vasculogenic mimicry. Clin Exp Med. 13:143–147. 2013. View Article : Google Scholar : PubMed/NCBI
17	Li M, Pathak RR, Lopez-Rivera E, Friedman SL, Aguirre-Ghiso JA and Sikora AG: The In Ovo Chick Chorioallantoic Membrane (CAM) Assay as an Efficient Xenograft Model of Hepatocellular Carcinoma. J Vis Exp. 2015:e52411. 2015.
18	Comşa Ş, Popescu R, Avram Ş, Ceaușu RA, Cîmpean AM and Raica M: Bevacizumab Modulation of the Interaction Between the MCF-7 Cell Line and the Chick Embryo Chorioallantoic Membrane. In Vivo. 31:199–203. 2017. View Article : Google Scholar : PubMed/NCBI
19	Workman P, Aboagye EO, Balkwill F, Balmain A, Bruder G, Chaplin DJ, Double JA, Everitt J, Farningham DA, Glennie MJ, et al: Committee of the National Cancer Research Institute: Guidelines for the welfare and use of animals in cancer research. Br J Cancer. 102:1555–1577. 2010. View Article : Google Scholar : PubMed/NCBI
20	Ha L, Noonan FP, De Fabo EC and Merlino G: Animal models of melanoma. J Investig Dermatol Symp Proc. 10:86–88. 2005. View Article : Google Scholar : PubMed/NCBI
21	Becker JC, Houben R, Schrama D, Voigt H, Ugurel S and Reisfeld RA: Mouse models for melanoma: A personal perspective. Exp Dermatol. 19:157–164. 2010. View Article : Google Scholar : PubMed/NCBI
22	Ribatti D: The chick embryo chorioallantoic membrane in the study of tumor angiogenesis. Rom J Morphol Embryol. 49:131–135. 2008.PubMed/NCBI
23	Mena S, Rodriguez ML, Ortega A, Priego S, Obrador E, Asensi M, Petschen I, Cerdá M, Brown BD and Estrela JM: Glutathione and Bcl-2 targeting facilitates elimination by chemoradiotherapy of human A375 melanoma xenografts overexpressing bcl-xl, bcl-2, and mcl-1. J Transl Med. 10:82012. View Article : Google Scholar : PubMed/NCBI
24	Rigon RB, Oyafuso MH, Fujimura AT, Gonçalez ML, do Prado AH, Gremião MP and Chorilli M: Nanotechnology-Based Drug Delivery Systems for Melanoma Antitumoral Therapy: A Review. BioMed Res Int. 2015:8418172015. View Article : Google Scholar : PubMed/NCBI
25	Niezgoda A, Niezgoda P and Czajkowski R: Novel Approaches to Treatment of Advanced Melanoma: A Review on Targeted Therapy and Immunotherapy. BioMed Res Int. 2015:8513872015. View Article : Google Scholar : PubMed/NCBI
26	Baderca F, Solovan C and Boghian L: Epidemiological and morphological data of ocular melanocytic lesions. Rom J Morphol Embryol. 54:77–83. 2013.PubMed/NCBI
27	Zurac S, Neagu M, Constantin C, Cioplea M, Nedelcu R, Bastian A, Popp C, Nichita L, Andrei R, Tebeica T, et al: Variations in the expression of TIMP1, TIMP2 and TIMP3 in cutaneous melanoma with regression and their possible function as prognostic predictors. Oncol Lett. 11:3354–3360. 2016.PubMed/NCBI
28	Wang L, Hurley DG, Watkins W, Araki H, Tamada Y, Muthukaruppan A, Ranjard L, Derkac E, Imoto S, Miyano S, et al: Cell cycle gene networks are associated with melanoma prognosis. PLoS One. 7:e342472012. View Article : Google Scholar : PubMed/NCBI
29	Fedorenko IV, Gibney GT and Smalley KSM: NRAS mutant melanoma: Biological behavior and future strategies for therapeutic management. Oncogene. 32:3009–3018. 2013. View Article : Google Scholar : PubMed/NCBI
30	Rodrigueza WV, Woolliscroft MJ, Ebrahim AS, Forgey R, McGovren PJ, Endert G, Wagner A, Holewa D, Aboukameel A, Gill RD, et al: Development and antitumor activity of a BCL-2 targeted single-stranded DNA oligonucleotide. Cancer Chemother Pharmacol. 74:151–166. 2014. View Article : Google Scholar : PubMed/NCBI
31	Russo AE, Torrisi E, Bevelacqua Y, Perrotta R, Libra M, McCubrey JA, Spandidos DA, Stivala F and Malaponte G: Melanoma: Molecular pathogenesis and emerging target therapies (Review). Int J Oncol. 34:1481–1489. 2009.PubMed/NCBI
32	Jones V and Katiyar SK: Emerging phytochemicals for prevention of melanoma invasion. Cancer Lett. 335:251–258. 2013. View Article : Google Scholar : PubMed/NCBI
33	Ribatti D, Annese T and Longo V: Angiogenesis and melanoma. Cancers (Basel). 2:114–132. 2010. View Article : Google Scholar : PubMed/NCBI
34	Streit M and Detmar M: Angiogenesis, lymphangiogenesis, and melanoma metastasis. Oncogene. 22:3172–3179. 2003. View Article : Google Scholar : PubMed/NCBI
35	Rozenberg GI, Monahan KB, Torrice C, Bear JE and Sharpless NE: Metastasis in an orthotopic murine model of melanoma is independent of RAS/RAF mutation. Melanoma Res. 20:361–371. 2010.PubMed/NCBI
36	Folkman J and Cotran R: Relation of vascular proliferation to tumor growth. Int Rev Exp Pathol. 16:207–248. 1976.PubMed/NCBI
37	Friend JV, Crevel RW, Williams TC and Parish WE: Immaturity of the inflammatory response of the chick chorioallantoic membrane. Toxicol In Vitro. 4:324–326. 1990. View Article : Google Scholar : PubMed/NCBI
38	Dupertuis YM, Delie F, Cohen M and Pichard C: In ovo method for evaluating the effect of nutritional therapies on tumor development, growth and vascularization. Clin Nutr Exp. 2:9–17. 2015. View Article : Google Scholar
39	Dehelean CA, Feflea S, Ganta S and Amiji M: Anti-angiogenic effects of betulinic acid administered in nanoemulsion formulation using chorioallantoic membrane assay. J Biomed Nanotechnol. 7:317–324. 2011. View Article : Google Scholar : PubMed/NCBI
40	Dehelean CA, Feflea S, Gheorgheosu D, Ganta S, Cimpean AM, Muntean D and Amiji MM: Anti-angiogenic and anti-cancer evaluation of betulin nanoemulsion in chicken chorioallantoic membrane and skin carcinoma in Balb/c mice. J Biomed Nanotechnol. 9:577–589. 2013. View Article : Google Scholar : PubMed/NCBI
41	Papoutsi M, Siemeister G, Weindel K, Tomarev SI, Kurz H, Schächtele C, Martiny-Baron G, Christ B, Marmé D and Wilting J: Active interaction of human A375 melanoma cells with the lymphatics in vivo. Histochem Cell Biol. 114:373–385. 2000.PubMed/NCBI
42	Papoutsi M, Kurz H, Schächtele C, Marmé D, Christ B, Pröls F and Wilting J: Induction of the blood-brain barrier marker neurothelin/HT7 in endothelial cells by a variety of tumors in chick embryos. Histochem Cell Biol. 113:105–113. 2000. View Article : Google Scholar : PubMed/NCBI
43	Marton A, Kúsz E, Kolozsi C, Tubak V, Zagotto G, Buzás K, Quintieri L and Vizler C: Vanillin Analogues o-Vanillin and 2,4,6-Trihydroxybenzaldehyde Inhibit NFĸB Activation and Suppress Growth of A375 Human Melanoma. Anticancer Res. 36:5743–5750. 2016. View Article : Google Scholar : PubMed/NCBI
44	Nowak-Sliwinska P, Segura T and Iruela-Arispe ML: The chicken chorioallantoic membrane model in biology, medicine and bioengineering. Angiogenesis. 17:779–804. 2014. View Article : Google Scholar : PubMed/NCBI
45	Beaumont KA, Mohana-Kumaran N and Haass NK: Modeling Melanoma In Vitro and In Vivo. Healthcare (Basel). 2:27–46. 2013. View Article : Google Scholar : PubMed/NCBI
46	Hu T, Zhang C, Tang Q, Su Y, Li B, Chen L, Zhang Z, Cai T and Zhu Y: Variant G6PD levels promote tumor cell proliferation or apoptosis via the STAT3/5 pathway in the human melanoma xenograft mouse model. BMC Cancer. 13:2512013. View Article : Google Scholar : PubMed/NCBI
47	Zheng AW, Jia DD, Xia LM, Jin G, Wu H and Li T: Impact of carboplatin plus paclitaxel combined with endostar against A375 melanoma cells: An in vitro and in vivo analysis. Biomed Pharmacother. 83:1321–1326. 2016. View Article : Google Scholar : PubMed/NCBI
48	Maishi N, Ohba Y, Akiyama K, Ohga N, Hamada J, Nagao-Kitamoto H, Alam MT, Yamamoto K, Kawamoto T, Inoue N, et al: Tumour endothelial cells in high metastatic tumours promote metastasis via epigenetic dysregulation of biglycan. Sci Rep. 6:280392016. View Article : Google Scholar : PubMed/NCBI
49	Jin J, Zhang Y, Li Y, Zhang H, Li H, Yuan X, Li X, Zhou W, Xu B, Zhang C, et al: RNA-interference-mediated downregulation of Pin1 suppresses tumorigenicity of malignant melanoma A375 cells. Neoplasma. 60:92–100. 2013. View Article : Google Scholar : PubMed/NCBI
50	Belizario JE: Immunodeficient Mouse Models: An Overview. Open Immunol J. 2:79–85. 2009. View Article : Google Scholar
51	Envigo: Research Models and Services: Oncology-Mutant Mice. Athymic Nude Mice. Envigo RMS Division. (Indianapolis, IN). 2016.http://www.envigo.com/resources/data-sheets/envigo-5193-131-leaflets-2016-nude-mouse-combination-a4-refs.pdf
52	Herwig N, Belter B and Pietzsch J: Extracellular S100A4 affects endothelial cell integrity and stimulates transmigration of A375 melanoma cells. Biochem Biophys Res Commun. 477:963–969. 2016. View Article : Google Scholar : PubMed/NCBI
53	Clark AG and Vignjevic DM: Modes of cancer cell invasion and the role of the microenvironment. Curr Opin Cell Biol. 36:13–22. 2015. View Article : Google Scholar : PubMed/NCBI
54	Craft N, Bruhn KW, Nguyen BD, Prins R, Liau LM, Collisson EA, De A, Kolodney MS, Gambhir SS and Miller JF: Bioluminescent imaging of melanoma in live mice. J Invest Dermatol. 125:159–165. 2005. View Article : Google Scholar : PubMed/NCBI
55	Gajda M and Kaminska-Winciorek G: Do not let to be late: Overview of reasons for melanoma delayed diagnosis. Asian Pac J Cancer Prev. 15:3873–3877. 2014. View Article : Google Scholar : PubMed/NCBI
56	Dehelean CA, Soica C, Pinzaru I, Coricovac D, Danciu C, Pavel I, Borcan F, Spandidos DA, Tsatsakis AM and Baderca F: Sex differences and pathology status correlated to the toxicity of some common carcinogens in experimental skin carcinoma. Food Chem Toxicol. 95:149–158. 2016. View Article : Google Scholar : PubMed/NCBI