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Article

Preparation and functional characterization of human vascular endothelial growth factor-melittin fusion protein with analysis of the antitumor activity in vitro and in vivo

  • Authors:
    • Dingding Wang
    • Lili Hu
    • Manman Su
    • Ju Wang
    • Tianmin Xu
  • View Affiliations / Copyright

    Affiliations: Department of Biotechnology, College of Life Science and Bio-pharmaceuticals, Guangdong Pharmaceutical University, Guangzhou, Guangdong, P.R. China, Department of Regenerative Medicine, College of Pharmacy, Jilin University, Changchun, Jilin, P.R. China, Guangdong Provincial Key Laboratory of Bio-engineering Medicine (National Engineering Research Centre of Genetic Medicine), Guangzhou, Guangdong, P.R. China, Department of Obstetrics and Gynecology, The Second Clinical Hospital, Jilin University, Changchun, Jilin, P.R. China
  • Pages: 1160-1168
    |
    Published online on: July 9, 2015
       https://doi.org/10.3892/ijo.2015.3078
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Abstract

Vascular endothelial growth factor and its tyrosine kinase receptors have been identified as key mediators of the regulation of pathologic blood vessel growth and maintenance in the promotion of angiogenesis and tumor growth. Therefore, an alternative approach to destroying tumor endothelium would be to make this tissue particularly sensitive to VEGF-mediated drug delivery. To verify this hypothesis, we generated a protein containing VEGF165 fused to melittin. Melittin is a small linear peptide composed of 26 amino acid residues that can exert toxic or inhibitory effects on many types of tumor cells. This protein is a cytolytic peptide that attacks lipid membranes, leading to significant toxicity. In the present study, the Pichia pastoris expression system was used to express the fusion protein. Under optimal conditions, stable VEGF165-melittin production was achieved using a series of purification steps. The activity of VEGF165-melittin fusion protein was compared with melittin for its ability to suppress the growth of tumor cell line in vitro. The fusion toxin selectively inhibited growth of human hepatocellular carcinoma HepG-2 cell line with high expression of VEGFR-2. We found that sensitivity of VEGFR-2 transfected 293 cells to VEGF165-melittin enhanced as the cellular VEGFR-2 density increased. In an in vivo initial experiment, the fusion protein inhibited tumor growth in xenografts assays. Furthermore, successful expression and characterization of the fusion protein demonstrated its efficacy for use as a novel treatment strategy for cancer.
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1 

Carmeliet P and Jain RK: Angiogenesis in cancer and other diseases. Nature. 407:249–257. 2000. View Article : Google Scholar : PubMed/NCBI

2 

Zhu W, Kato Y and Artemov D: Heterogeneity of tumor vasculature and antiangiogenic intervention: Insights from MR angiography and DCE-MRI. PLoS One. 9:e865832014. View Article : Google Scholar : PubMed/NCBI

3 

Ferrara N and Ferrara N: Vascular endothelial growth factor: Molecular and biological aspects. Curr Top Microbiol Immunol. 237:1–30. 1999.PubMed/NCBI

4 

Hicklin DJ and Ellis LM: Role of the vascular endothelial growth factor pathway in tumor growth and angiogenesis. J Clin Oncol. 23:1011–1027. 2005. View Article : Google Scholar

5 

Zhang D, Li B, Shi J, Zhao L, Zhang X, Wang C, Hou S, Qian W, Kou G, Wang H, et al: Suppression of tumor growth and metastasis by simultaneously blocking vascular endothelial growth factor (VEGF)-A and VEGF-C with a receptor-immunoglobulin fusion protein. Cancer Res. 70:2495–2503. 2010. View Article : Google Scholar : PubMed/NCBI

6 

Chen AI and Advani RH: Beyond the guidelines in the treatment of peripheral T-cell lymphoma: New drug development. J Natl Compr Canc Netw. 6:428–435. 2008.PubMed/NCBI

7 

Tischer E, Mitchell R, Hartman T, Silva M, Gospodarowicz D, Fiddes JC and Abraham JA: The human gene for vascular endothelial growth factor. Multiple protein forms are encoded through alternative exon splicing. J Biol Chem. 266:11947–11954. 1991.PubMed/NCBI

8 

Houck KA, Leung DW, Rowland AM, Winer J and Ferrara N: Dual regulation of vascular endothelial growth factor bioavailability by genetic and proteolytic mechanisms. J Biol Chem. 267:26031–26037. 1992.PubMed/NCBI

9 

Koutsioumpa M, Poimenidi E, Pantazaka E, Theodoropoulou C, Skoura A, Megalooikonomou V, Kieffer N, Courty J, Mizumoto S, Sugahara K, et al: Receptor protein tyrosine phosphatase beta/zeta is a functional binding partner for vascular endothelial growth factor. Mol Cancer. 14:19–34. 2015. View Article : Google Scholar : PubMed/NCBI

10 

Oršolić N: Bee venom in cancer therapy. Cancer Metastasis Rev. 31:173–194. 2012. View Article : Google Scholar

11 

Wang C, Chen T, Zhang N, Yang M, Li B, Lü X, Cao X and Ling C: Melittin, a major component of bee venom, sensitizes human hepatocellular carcinoma cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis by activating CaMKII-TAK1-JNK/p38 and inhibiting IκBα kinase-NF-κB. J Biol Chem. 284:3804–3813. 2009. View Article : Google Scholar

12 

Gajski G and Garaj-Vrhovac V: Melittin: A lytic peptide with anti-cancer properties. Environ Toxicol Pharmacol. 36:697–705. 2013. View Article : Google Scholar : PubMed/NCBI

13 

Liu S, Yu M, He Y, Xiao L, Wang F, Song C, Sun S, Ling C and Xu Z: Melittin prevents liver cancer cell metastasis through inhibition of the Rac1-dependent pathway. Hepatology. 47:1964–1973. 2008. View Article : Google Scholar : PubMed/NCBI

14 

Hoskin DW and Ramamoorthy A: Studies on anticancer activities of antimicrobial peptides. Biochim Biophys Acta. 1778:357–375. 2008. View Article : Google Scholar

15 

Soman NR, Baldwin SL, Hu G, Marsh JN, Lanza GM, Heuser JE, Arbeit JM, Wickline SA and Schlesinger PH: Molecularly targeted nanocarriers deliver the cytolytic peptide melittin specifically to tumor cells in mice, reducing tumor growth. J Clin Invest. 119:2830–2842. 2009. View Article : Google Scholar : PubMed/NCBI

16 

Lee MT, Chen FY and Huang HW: Molecular mechanism of Peptide-induced pores in membranes. Biochemistry. 43:3590–3599. 2004. View Article : Google Scholar : PubMed/NCBI

17 

Huh JE, Kang JW, Nam D, Baek YH, Choi DY, Park DS and Lee JD: Melittin suppresses VEGF-A-induced tumor growth by blocking VEGFR-2 and the COX-2-mediated MAPK signaling pathway. J Nat Prod. 75:1922–1929. 2012. View Article : Google Scholar : PubMed/NCBI

18 

Shin JM, Jeong YJ, Cho HJ, Park KK, Chung IK, Lee IK, Kwak JY, Chang HW, Kim CH, Moon SK, et al: Melittin suppresses HIF-1α/VEGF expression through inhibition of ERK and mTOR/p70S6K pathway in human cervical carcinoma cells. PLoS One. 8:e693802013. View Article : Google Scholar

19 

Pichia Expression Kit Version F. A Manual of Methods for Expression of Recombinant Proteins in Pichia pastoris. Invitrogen. 2002

20 

Ciomber A, Smagur A, Mitrus I, Cichoń T, Smolarczyk R, Sochanik A, Szala S and Jarosz M: Antitumor effects of recombinant antivascular protein ABRaA-VEGF121 combined with IL-12 gene therapy. Arch Immunol Ther Exp (Warsz). 62:161–168. 2014. View Article : Google Scholar

21 

Xu WW, Li B, Lam AK, Tsao SW, Law SY, Chan KW, Yuan QJ and Cheung AL: Targeting VEGFR1- and VEGFR2-expressing non-tumor cells is essential for esophageal cancer therapy. Oncotarget. 6:1790–1805. 2015.PubMed/NCBI

22 

Liu M, Zong J, Liu Z, Li L, Zheng X, Wang B and Sun G: A novel melittin-MhIL-2 fusion protein inhibits the growth of human ovarian cancer SKOV3 cells in vitro and in vivo tumor growth. Cancer Immunol Immunother. 62:889–895. 2013. View Article : Google Scholar : PubMed/NCBI

23 

Lee MT, Sun TL, Hung WC and Huang HW: Process of inducing pores in membranes by melittin. Proc Natl Acad Sci USA. 110:14243–14248. 2013. View Article : Google Scholar : PubMed/NCBI

24 

Sommer A, Fries A, Cornelsen I, Speck N, Koch-Nolte F, Gimpl G, Andrä J, Bhakdi S and Reiss K: Melittin modulates keratinocyte function through P2 receptor-dependent ADAM activation. J Biol Chem. 287:23678–23689. 2012. View Article : Google Scholar : PubMed/NCBI

25 

Qu L, Jiang M, Li Z, Pu F, Gong L, Sun L, Gong R, Ji G and Si J: Inhibitory effect of biosynthetic nanoscale peptide Melittin on hepatocellular carcinoma, driven by survivin promoter. J Biomed Nanotechnol. 10:695–706. 2014. View Article : Google Scholar : PubMed/NCBI

26 

Jo M, Park MH, Kollipara PS, An BJ, Song HS, Han SB, Kim JH, Song MJ and Hong JT: Anti-cancer effect of bee venom toxin and melittin in ovarian cancer cells through induction of death receptors and inhibition of JAK2/STAT3 pathway. Toxicol Appl Pharmacol. 258:72–81. 2012. View Article : Google Scholar

27 

Liu H, Han Y, Fu H, Liu M, Wu J, Chen X, Zhang S and Chen Y: Construction and expression of sTRAIL-melittin combining enhanced anticancer activity with antibacterial activity in Escherichia coli. Appl Microbiol Biotechnol. 97:2877–2884. 2013. View Article : Google Scholar

28 

Su M, Chang W, Cui M, Lin Y, Wu S and Xu T: Expression and anticancer activity analysis of recombinant human uPA1-43-melittin. Int J Oncol. 46:619–626. 2015.

29 

Wang DD, Su MM, Sun Y, Huang SL, Wang J and Yan WQ: Expression, purification and characterization of a human single-chain Fv antibody fragment fused with the Fc of an IgG1 targeting a rabies antigen in Pichia pastoris. Protein Expr Purif. 86:75–81. 2012. View Article : Google Scholar : PubMed/NCBI

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Copy and paste a formatted citation
Spandidos Publications style
Wang D, Hu L, Su M, Wang J and Xu T: Preparation and functional characterization of human vascular endothelial growth factor-melittin fusion protein with analysis of the antitumor activity in vitro and in vivo. Int J Oncol 47: 1160-1168, 2015.
APA
Wang, D., Hu, L., Su, M., Wang, J., & Xu, T. (2015). Preparation and functional characterization of human vascular endothelial growth factor-melittin fusion protein with analysis of the antitumor activity in vitro and in vivo. International Journal of Oncology, 47, 1160-1168. https://doi.org/10.3892/ijo.2015.3078
MLA
Wang, D., Hu, L., Su, M., Wang, J., Xu, T."Preparation and functional characterization of human vascular endothelial growth factor-melittin fusion protein with analysis of the antitumor activity in vitro and in vivo". International Journal of Oncology 47.3 (2015): 1160-1168.
Chicago
Wang, D., Hu, L., Su, M., Wang, J., Xu, T."Preparation and functional characterization of human vascular endothelial growth factor-melittin fusion protein with analysis of the antitumor activity in vitro and in vivo". International Journal of Oncology 47, no. 3 (2015): 1160-1168. https://doi.org/10.3892/ijo.2015.3078
Copy and paste a formatted citation
x
Spandidos Publications style
Wang D, Hu L, Su M, Wang J and Xu T: Preparation and functional characterization of human vascular endothelial growth factor-melittin fusion protein with analysis of the antitumor activity in vitro and in vivo. Int J Oncol 47: 1160-1168, 2015.
APA
Wang, D., Hu, L., Su, M., Wang, J., & Xu, T. (2015). Preparation and functional characterization of human vascular endothelial growth factor-melittin fusion protein with analysis of the antitumor activity in vitro and in vivo. International Journal of Oncology, 47, 1160-1168. https://doi.org/10.3892/ijo.2015.3078
MLA
Wang, D., Hu, L., Su, M., Wang, J., Xu, T."Preparation and functional characterization of human vascular endothelial growth factor-melittin fusion protein with analysis of the antitumor activity in vitro and in vivo". International Journal of Oncology 47.3 (2015): 1160-1168.
Chicago
Wang, D., Hu, L., Su, M., Wang, J., Xu, T."Preparation and functional characterization of human vascular endothelial growth factor-melittin fusion protein with analysis of the antitumor activity in vitro and in vivo". International Journal of Oncology 47, no. 3 (2015): 1160-1168. https://doi.org/10.3892/ijo.2015.3078
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