The biological effect and mechanism of the Wnt/β‑catenin signaling pathway on malignant melanoma A375 cells

Lin,Yuxia; Wang,Fangfei; Xing,Qingfei; Guo,Feng; Wang,Mengzhen; Li,Yunjie

doi:10.3892/etm.2018.6413

The biological effect and mechanism of the Wnt/β‑catenin signaling pathway on malignant melanoma A375 cells

Authors:
- Yuxia Lin
- Fangfei Wang
- Qingfei Xing
- Feng Guo
- Mengzhen Wang
- Yunjie Li
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Affiliations: Department of Urology, Jinan Central Hospital, Jinan, Shandong 250013, P.R. China, Department of Finance, Jinan Central Hospital, Jinan, Shandong 250013, P.R. China, Department of Equipment, Jinan Central Hospital, Jinan, Shandong 250013, P.R. China
Published online on: July 6, 2018 https://doi.org/10.3892/etm.2018.6413
Pages: 2032-2037

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Abstract

The present study aimed to determine the influence of the Wnt/β-catenin signaling pathway on the proliferation, invasion, migration and apoptosis of malignant melanoma (MM) A375 cells. β‑catenin interfering lentivirus liquid (β‑catenin‑RNAi‑LV) and empty vector lentivirus liquid (β‑catenin‑negative‑LV) were used to infect A375 cells. Infected cells were obtained and marked as A375‑RNA interference (A375‑RNAi) or A375‑negative, respectively. Western blotting was used to measure the expression of β‑catenin in infected cells and uninfected cells were utilized as a control. An MTT assay was adopted to measure cell proliferation and the clone formation of cells was assessed. In addition, the Transwell method was used to detect cell invasion and migration in vitro and flow cytometry was utilized to determine cell apoptosis. Western blot analysis demonstrated that β‑catenin was highly expressed in uninfected A375 cells but exhibited reduced expression in A375‑RNAi cells. These results indicate that β‑catenin expression is effectively silenced by β‑catenin‑RNAi‑LV. The proliferative and clone forming abilities of A375‑RNAi cells were impaired compared with A375‑negative and A375 cells. Additionally, the apoptosis rate was increased and the invasion and migration of A375‑RNAi cells was decreased. However, no significant differences were identified in the proliferation, clone formation, apoptosis rate, invasion and migration of A375‑negative cells compared with A375 cells. Therefore, the current study demonstrated that the inhibition of β‑catenin expression or activity inhibits cell proliferation and invasion and migration, further downregulating the expression of anti‑apoptotic genes and accelerating cellular apoptosis.

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1	Erdmann F, Lortettieulent J, Schüz J, Zeeb H, Greinert R, Breitbart EW and Bray F: International trends in the incidence of malignant melanoma 1953–2008-are recent generations at higher or lower risk? Int J Cancer. 132:385–400. 2013. View Article : Google Scholar : PubMed/NCBI
2	Chang JW, Yeh KY, Wang CH, Yang TS, Chiang HF, Wei FC, Kuo TT and Yang CH: Malignant melanoma in Taiwan: A prognostic study of 181 cases. Melanoma Res. 14:537–541. 2004. View Article : Google Scholar : PubMed/NCBI
3	Perlis C and Herlyn M: Recent advances in melanoma biology. Oncologist. 9:182–187. 2004. View Article : Google Scholar : PubMed/NCBI
4	Miao Y, Owen NK, Whitener D, Gallazzi F, Hoffman TJ and Quinn TP: In vivo evaluation of 188Re-labeled alpha-melanocyte stimulating hormone peptide analogs for melanoma therapy. Int J Cancer. 101:480–187. 2002. View Article : Google Scholar : PubMed/NCBI
5	Delmas V, Beermann F, Martinozzi S, Carreira S, Ackermann J, Kumasaka M, Denat L, Goodall J, Luciani F, Viros A, et al: Beta-catenin induces immortalization of melanocytes by suppressing p16INK4a expression and cooperates with N-Ras in melanoma development. Genes Dev. 21:2923–2935. 2007. View Article : Google Scholar : PubMed/NCBI
6	Chin L, Garraway LA and Fisher DE: Malignant melanoma: Genetics and therapeutics in the genomic era. Genes Dev. 20:2149–2182. 2006. View Article : Google Scholar : PubMed/NCBI
7	Lens MB, Dawes M, Goodacre T and Bishop JA: Excision margins in the treatment of primary cutaneous melanoma: A systematic review of randomized controlled trials comparing narrow vs. wide excision. Arch Surg. 137:1101–1105. 2002. View Article : Google Scholar : PubMed/NCBI
8	Zhou Qian QH, Yao X, Zhu M and Jiang Y: Clinical pathological analysis of 29 cases of cutaneous malignant melanoma. Guide of China Medicine. 6:20–22. 2008.(In Chinese).
9	Chien AJ, Moore EC, Lonsdorf AS, Kulikauskas RM, Rothberg BG, Berger AJ, Major MB, Hwang ST, Rimm DL and Moon RT: Activated Wnt/ß-catenin signaling in melanoma is associated with decreased proliferation in patient tumors and a murine melanoma model. Proc Natl Acad Sci USA. 106:1193–1198. 2009. View Article : Google Scholar : PubMed/NCBI
10	Ciolczyk-Wierzbicka D, Gil D and Laidler P: The inhibition of cell proliferation using silencing of N-cadherin gene by siRNA process in human melanoma cell lines. Curr Med Chem. 19:145–151. 2012. View Article : Google Scholar : PubMed/NCBI
11	Niu LL and Hao J: Rosiglitazone inhibition of A375 human malignant melanoma cell invasion. Chinese J Dermatol. 42:831–834. 2009.
12	Riccardo F, Iussich S, Maniscalco L, Mayayo SL, Rosa GL, Arigoni M, Maria RD, Gattino F, Lanzardo S, Lardone E, et al: CSPG4-specific immunity and survival prolongation in dogs with oral malignant melanoma immunized with human CSPG4 DNA. Clin Cancer Res. 20:3753–3762. 2014. View Article : Google Scholar : PubMed/NCBI
13	Rappa G, Mercapide J, Anzanello F, Le TT, Mary G, Johlfs RRF, Wilsch-Bräuninger M, Corbeil D and Loricoa A: Wnt interaction and extracellular release of prominin-1/CD133 in human malignant melanoma cells. Exp Cell Res. 319:810–819. 2013. View Article : Google Scholar : PubMed/NCBI
14	Tarapore RS, Siddiqui IA, Saleem M, Spiegelman V and Mukhtar H: Abstract 3794: Growth inhibition of human melanoma cells in vitro and in vivo by lupeol is associated with inhibition of Wnt/β-catenin signaling. Cancer Res. 70 Suppl:S37942010. View Article : Google Scholar
15	Katiyar SK and Vaid M: Abstract 3683: Bioactive phytochemical proanthocyanidins target β-catenin signaling in preventing invasive potential of human melanoma cells. Cancer Res. 73 Suppl:S36832013. View Article : Google Scholar
16	Widlund HR, Horstmann MA, Price ER, Cui J, Lessnick SL, Wu M, He X and Fisher DE: β-Catenin-induced melanoma growth requires the downstream target Microphthalmia-associated transcription factor. J Cell Biol. 158:1079–1087. 2002. View Article : Google Scholar : PubMed/NCBI
17	Beck D, Niessner H, Krieg K, Gogel J, Bonin M, Garbe C and Meier F: Chemosensitizing activity of the mTOR inhibitor temsirolimus (Torisel) in metastatic melanoma involves DKK1. Journal der Deutschen Dermatologischen Gesellschaft. 11:6. 2013.
18	Mikheev AM, Mikheeva SA, Rostomily R and Zarbl H: Dickkopf-1 activates cell death in MDA-MB435 melanoma cells. Biochem Biophys Res Commun. 352:675–680. 2007. View Article : Google Scholar : PubMed/NCBI
19	Pawlikowski JS, Mcbryan T, Van TJ, Drotar ME, Hewitt RN, Maier AB, King A, Blyth K, Wu H and Adams PD: Wnt signaling potentiates nevogenesis. Proc Natl Acad Sci USA. 110:16009–16014. 2013. View Article : Google Scholar : PubMed/NCBI
20	Mælandsmo GM, Holm R, Nesland JM, Fodstad Ø and Flørenes VA: Reduced β-catenin expression in the cytoplasm of advanced-stage superficial spreading malignant melanoma. Clin Cancer Res J Am Assoc Cancer Res. 9:3383–3388. 2003.
21	Cimetta E, Cannizzaro C, James R, Biechele T, Moon RT, Elvassore N and Vunjak-Novakovic G: Microfluidi device generating stable concentration gradients for long term cell culture: Application to Wnt3a regulation of β-catenin signaling. Lab Chip. 10:3277–3283. 2010. View Article : Google Scholar : PubMed/NCBI
22	Comodo AN, Bachi ALL, Soares MF, Franco M, de Paulo V and Teixeira C: Galectin-3 expression favors metastasis in murine melanoma. Adv Biosci Biotechnol. 4:55–62. 2013. View Article : Google Scholar
23	Shah PK, Walker MP, Sims CE, Major MB and Allbritton NL: Dynamics and evolution of β-catenin-dependent Wnt signaling revealed through massively parallel clonogenic screening. Integr Biol. 6:673–684. 2014. View Article : Google Scholar
24	Tobias S, Moritz M, Daniel E and Birgit S, Michael S, Martin S, Claus G and Birgit S: β-catenin signaling increases during melanoma progression and promotes tumor cell survival and chemoresistance. PloS One. 6:e234292011. View Article : Google Scholar : PubMed/NCBI
25	Dong F and Herlyn M: The dynamic roles of cell-surface receptors in melanoma development. From Melanocytes to Melanoma. 169–181. 2006.
26	Hsu MY, Ling L and Herlyn M: Cultivation of normal human epidermal melanocytes in the absence of phorbol esters. Methods Mol Med. 107:13–28. 2005.PubMed/NCBI
27	Tucci MG, Lucarini G, Brancorsini D, Zizzi A, Pugnaloni A, Giacchetti A, Ricotti G and Biagini G: Involvement of E-cadherin, β-catenin, Cdc42 and CXCR4 in the progression and prognosis of cutaneous melanoma. Br J Dermatol. 157:1212–1216. 2007. View Article : Google Scholar : PubMed/NCBI
28	Kuno T, Tsukamoto T, Hara A and Tanaka T: Cancer chemoprevention through the induction of apoptosis by natural compounds. J Biophys Chem. 3:156–173. 2012. View Article : Google Scholar
29	Novellino L, De FA, Deho P, Perrone F, Pilotti S, Parmiani G and Castelli C: PTPRK negatively regulates transcriptional activity of wild type and mutated oncogenic beta-catenin and affects membrane distribution of beta-catenin/E-cadherin complexes in cancer cells. Cell Signal. 20:872–883. 2008. View Article : Google Scholar : PubMed/NCBI
30	Lee DJ, Kang DH, Choi M, Choi YJ, Lee JY, Park JH, Park YJ, Lee KW and Kang SW: Peroxiredoxin-2 represses melanoma metastasis by increasing E-Cadherin/β-Catenin complexes in adherens junctions. Cancer Res. 73:4744–4757. 2013. View Article : Google Scholar : PubMed/NCBI
31	Pećina-Šlaus N, Žigmund M, Kušec V, Martić TN, Čačić M and Šlaus M: E-cadherin and β-catenin expression patterns in malignant melanoma assessed by image analysis. J Cutan Pathol. 34:239–246. 2007. View Article : Google Scholar : PubMed/NCBI
32	Vaid M, Prasad R, Sun Q and Katiyar SK: Silymarin targets β-catenin signaling in blocking migration/invasion of human melanoma cells. PloS One. 6:e230002010. View Article : Google Scholar
33	Damsky WE, Curley DP, Santhanakrishnan M, Rosenbaum LE, Platt JT, Rothberg Gould BE, Taketo MM, Dankort D, Rimm DL, McMahon M and Bosenberg M: β-catenin signaling controls metastasis in Braf-activated Pten-deficient melanomas. Cancer Cell. 20:741–754. 2011. View Article : Google Scholar : PubMed/NCBI
34	Chon E, Thompson V, Schmid S and Stein TJ: Activation of the canonical Wnt/β-catenin signalling pathway is rare in canine malignant melanoma tissue and cell lines. J Comp Pathol. 148:178–187. 2013. View Article : Google Scholar : PubMed/NCBI
35	Dang CV, O'Donnell KA, Zeller KI, Nguyen T, Osthusa RC and Lia F: The c-Myc target gene network. Semin Cancer Biol. 16:253–264. 2006. View Article : Google Scholar : PubMed/NCBI
36	Zeller KI, Zhao X, Lee CWH, Chiu KP, Yao F, Yustein JT, Ooi HS, Orlov YL, Shahab A, Yong HC, et al: Global mapping of c-Myc binding sites and target gene networks in human B cells. Proc Natl Acad Sci USA. 103:17834–17839. 2006. View Article : Google Scholar : PubMed/NCBI
37	Schittek B and Sinnberg T: Biological functions of casein kinase 1 isoforms and putative roles in tumorigenesis. Mol Cancer. 13:2312014. View Article : Google Scholar : PubMed/NCBI