Islet-1 promotes the proliferation and invasion, and inhibits the apoptosis of A375 human melanoma cells

Zhu,Xiaoling; Li,Yuzhen; Meng,Qinggang

doi:10.3892/ijmm.2018.3569

Islet-1 promotes the proliferation and invasion, and inhibits the apoptosis of A375 human melanoma cells

Authors:
- Xiaoling Zhu
- Yuzhen Li
- Qinggang Meng
View Affiliations

Affiliations: Department of Dermatology, The First Hospital of Harbin, Harbin, Heilongjiang 150010, P.R. China, Department of Dermatology, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150081, P.R. China, Department of Orthopaedic Surgery, The First Hospital of Harbin, Harbin, Heilongjiang 150010, P.R. China
Published online on: March 14, 2018 https://doi.org/10.3892/ijmm.2018.3569
Pages: 3680-3690

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

The aim of this study was to examine the effects of the insulin gene enhancer-binding protein, islet-1 (ISL1), on the proliferation, invasion and apoptosis of the human melanoma cell line, A375. An ISL1 overexpression lentiviral vector was constructed and transfected into the A375 cells. The proliferation of the A375 cells transfected with the ISL1 vector (termed A375/ISL1 cells) was examined by MTT assay, flow cytometry and TUNEL assay, and cell invasion was examined by Transwell assay. The expression levels of matrix metalloproteinase (MMP)-2 and MMP-9 were measured by qPCR and western blot analysis; the expression levels of Akt and p-Akt were measured in the cells treated with vascular endothelial growth factor (VEGF) and the PI3K/Akt inhibitor, LY294002, by western blot analysis. The optical density value of the A375/ISL1 cells was increased after 12 h of culture (P<0.001), as shown by MTT assay. The ratio of apoptotic A375/ISL1 cells was significantly decreased (P<0.001), as shown by flow cytometry and TUNEL assay. In addition, the average penetration rate of the A375/ISL1 cells significantly increased (P<0.001), as shown by Transwell assay. The expression levels of MMP-2 and MMP-9 were significantly increased in the A375/ISL1 cells, as shown by qPCR and western blot analysis (P<0.001). Moreover, treatment of the A375/ISL1 cells with VEGF for 48 h increased the expression of Akt and p-Akt compared with the control cells transfected with A375/green fluorescent protein (GFP) (P<0.05; P<0.001, respectively). In addition, in the A375/ISL1 cells treated with the LY294002 inhibitor for 24 and 48 h, the level of Akt was also found to increase compared to the control A375/GFP cells (P<0.05). On the whole, the findings of this study indicate that the overexpression of ISL1 promotes the proliferation and invasion, and inhibits the apoptosis of A375 melanoma cells. ISL1 thus plays an important role in A375 cell survival, and these effects are possibly mediate via the PI3K/Akt signaling pathway.

View Figures

View References

1	Misu M, Ouji Y, Kawai N, Nishimura F, Nakamura-Uchiyama F and Yoshikawa M: Effects of Wnt-10b on proliferation and differentiation of murine melanoma cells. Biochem Biophys Res Commun. 463:618–623. 2015. View Article : Google Scholar : PubMed/NCBI
2	Raghunath A, Sambarey A, Sharma N, Mahadevan U and Chandra N: A molecular systems approach to modelling human skin pigmentation: identifying underlying pathways and critical components. BMC Res Notes. 8:1702015. View Article : Google Scholar : PubMed/NCBI
3	Siegel R, Ma J, Zou Z and Jemal A: Cancer statistics, 2014. CA Cancer J Clin. 64:9–29. 2014. View Article : Google Scholar : PubMed/NCBI
4	Karlsson O, Thor S, Norberg T, Ohlsson H and Edlund T: Insulin gene enhancer binding protein Isl-1 is a member of a novel class of proteins containing both a homeo- and a Cys-His domain. Nature. 344:879–882. 1990. View Article : Google Scholar : PubMed/NCBI
5	Nasif S, de Souza FS, González LE, Yamashita M, Orquera DP, Low MJ and Rubinstein M: Islet 1 specifies the identity of hypothalamic melanocortin neurons and is critical for normal food intake and adiposity in adulthood. Proc Natl Acad Sci USA. 112:E1861–E1870. 2015. View Article : Google Scholar : PubMed/NCBI
6	Cai CL, Liang X, Shi Y, Chu PH, Pfaff SL, Chen J and Evans S: Isl1 identifies a cardiac progenitor population that proliferates prior to differentiation and contributes a majority of cells to the heart. Dev Cell. 5:877–889. 2003. View Article : Google Scholar : PubMed/NCBI
7	Huber GF: Modern management of Merkel cell carcinoma. Curr Opin Otolaryngol Head Neck Surg. 22:109–115. 2014. View Article : Google Scholar : PubMed/NCBI
8	Agaimy A, Erlenbach-Wünsch K, Konukiewitz B, Schmitt AM, Rieker RJ, Vieth M, Kiesewetter F, Hartmann A, Zamboni G, Perren A, et al: ISL1 expression is not restricted to pancreatic well-differentiated neuroendocrine neoplasms, but is also commonly found in well and poorly differentiated neuroendocrine neoplasms of extrapancreatic origin. Mod Pathol. 26:995–1003. 2013. View Article : Google Scholar : PubMed/NCBI
9	Su X, Wang P, Wang X, Cao B, Li L and Liu Q: Apoptosis of U937 cells induced by hematoporphyrin monomethyl ether-mediated sonodynamic action. Cancer Biother Radiopharm. 28:207–217. 2013. View Article : Google Scholar : PubMed/NCBI
10	Hao Q, Li W, Zhang C, Qin X, Xue X, Li M, Shu Z, Xu T, Xu Y, Wang W, et al: TNFα induced FOXP3-NFκB interaction dampens the tumor suppressor role of FOXP3 in gastric cancer cells. Biochem Biophys Res Commun. 430:436–441. 2013. View Article : Google Scholar
11	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
12	Piérard-Franchimont C, Hermanns-Lê T, Delvenne P and Piérard GE: Dormancy of growth-stunted malignant melanoma: sustainable and smoldering patterns. Oncol Rev. 8:2522014. View Article : Google Scholar
13	Aris M and Barrio MM: Combining immunotherapy with oncogene-targeted therapy: a new road for melanoma treatment. Front Immunol. 6:462015. View Article : Google Scholar : PubMed/NCBI
14	Graham RP, Shrestha B, Caron BL, Smyrk TC, Grogg KL, Lloyd RV and Zhang L: Islet-1 is a sensitive but not entirely specific marker for pancreatic neuroendocrine neoplasms and their metastases. Am J Surg Pathol. 37:399–405. 2013. View Article : Google Scholar : PubMed/NCBI
15	Bu L, Jiang X, Martin-Puig S, Caron L, Zhu S, Shao Y, Roberts DJ, Huang PL, Domian IJ and Chien KR: Human ISL1 heart progenitors generate diverse multipotent cardiovascular cell lineages. Nature. 460:113–117. 2009. View Article : Google Scholar : PubMed/NCBI
16	Shi Y, Zhao S, Li J and Mao B: Islet-1 is required for ventral neuron survival in Xenopus. Biochem Biophys Res Commun. 388:506–510. 2009. View Article : Google Scholar : PubMed/NCBI
17	Guo T, Wang W, Zhang H, Liu Y, Chen P, Ma K and Zhou C: ISL1 promotes pancreatic islet cell proliferation. PLoS One. 6:e223872011. View Article : Google Scholar : PubMed/NCBI
18	Ediger BN, Du A, Liu J, Hunter CS, Walp ER, Schug J, Kaestner KH, Stein R, Stoffers DA and May CL: Islet-1 is essential for pancreatic β-cell function. Diabetes. 63:4206–4217. 2014. View Article : Google Scholar : PubMed/NCBI
19	Weinberger F, Mehrkens D, Starbatty J, Nicol P and Eschenhagen T: Assessment of DNA synthesis in Islet-1⁺ cells in the adult murine heart. Biochem Biophys Res Commun. 456:294–297. 2015. View Article : Google Scholar
20	Kilk K, Magzoub M, Pooga M, Eriksson LE, Langel U and Gräslund A: Cellular internalization of a cargo complex with a novel peptide derived from the third helix of the islet-1 homeodomain. Comparison with the penetratin peptide. Bioconjug Chem. 12:911–916. 2001. View Article : Google Scholar : PubMed/NCBI
21	Koo J, Mertens RB, Mirocha JM, Wang HL and Dhall D: Value of Islet 1 and PAX8 in identifying metastatic neuroendocrine tumors of pancreatic origin. Mod Pathol. 25:893–901. 2012. View Article : Google Scholar : PubMed/NCBI
22	Ferrara N, Carver-Moore K, Chen H, Dowd M, Lu L, O'Shea KS, Powell-Braxton L, Hillan KJ and Moore MW: Heterozygous embryonic lethality induced by targeted inactivation of the VEGF gene. Nature. 380:439–442. 1996. View Article : Google Scholar : PubMed/NCBI
23	Barzelay A, Ben-Shoshan J, Entin-Meer M, Maysel-Auslender S, Afek A, Barshack I, Keren G and George J: A potential role for islet-1 in post-natal angiogenesis and vasculogenesis. Thromb Haemost. 103:188–197. 2010. View Article : Google Scholar : PubMed/NCBI
24	Linning KD, Tai MH, Madhukar BV, Chang CC, Reed DN Jr, Ferber S, Trosko JE and Olson LK: Redox-mediated enrichment of self-renewing adult human pancreatic cells that possess endocrine differentiation potential. Pancreas. 29:e64–e76. 2004. View Article : Google Scholar : PubMed/NCBI