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01-05-2013 | Original article

A novel melittin-MhIL-2 fusion protein inhibits the growth of human ovarian cancer SKOV₃ cells in vitro and in vivo tumor growth

Authors: Mingjun Liu, Jinbao Zong, Zimin Liu, Ling Li, Xu Zheng, Bin Wang, Guirong Sun

Published in: Cancer Immunology, Immunotherapy | Issue 5/2013

Abstract

In the current study, we produced a novel fusion protein (melittin-mutant human interleukin 2, melittin-MhIL-2) comprising a mutant human interleukin 2 (Arg88/Ala125) genetically linked to melittin. The plasmid pET15b-melittin-MhIL-2 (Arg88/Ala125) was transformed into E. coli for protein expression. The expressed melittin-MhIL-2 protein was purified using a series of purification steps. The interleukin 2 (IL-2) activity of melittin-MhIL-2 fusion protein was compared with recombinant human interleukin 2 (rhIL-2) for its ability to induce CTLL-2 proliferation. Moreover, the fusion protein directly inhibits the growth of human ovarian cancer SKOV₃ cells in vitro. In an in vivo initial experiment, the fusion protein inhibited tumor growth in ovarian cancer mice. In conclusion, we generated a novel melittin-MhIL-2 fusion protein that retained functional activity of IL-2 and melittin and inhibited tumor growth in vivo.

Ramirez I, Chon HS, Apte SM (2011) The role of surgery in the management of epithelial ovarian cancer. Cancer Control 18:22–30PubMed

Al-Alem L, Southard RC, Kilgore MW, Curry TE (2011) Specific thiazolidinediones inhibit ovarian cancer cell line proliferation and cause cell cycle arrest in a PPARγ independent manner. PLoS ONE 6:e16179PubMedCrossRef

Al-Bahlani S, Fraser M, Wong AY, Sayan BSR, Melino G, Tsang BK (2011) P73 regulates cisplatin-induced apoptosis in ovarian cancer cells via a calcium/calpain-dependent mechanism. Oncogene 30:4219–4230PubMedCrossRef

Jain A, Dubashi B, Reddy KS, Jain P (2011) Weekly paclitaxel in ovarian cancer-the latest success story. Curr Oncol 18:16–17PubMedCrossRef

Reisfeld RA, Becker JC, Gillies SD (1997) Immunocytokines: a new approach to immunotherapy of melanoma. Melanoma Res 7(Suppl 2):S99–106PubMed

Rosenberg SA, Yang JC, White DE, Steinberg SM (1998) Durability of complete responses in patients with metastatic cancer treated with high-dose interleukin-2: identification of the antigens mediating response. Ann Surg 228:307–319PubMedCrossRef

Morgan DA, Ruscetti FW, Gallo R (1976) Selective in vitro growth of lymphocytes from normal human bone marrows. Science 193:1007–1008PubMedCrossRef

Henney CS, Kuribayashi K, Kern DE, Gillis S (1981) Interleukin 2 augments natural killer cell activity. Nature 291:335–338PubMedCrossRef

Ortaldo JR, Mason AT, Gerard JP, Henderson LE, Farrar W, Hopkins RF 3rd, Herberman RB, Rabin H (1984) Effects of natural and recombinant IL 2 on regulation of IFN gamma production and natural killer activity: lack of involvement of the Tac antigen for these immunoregulatory effects. J Immunol 133:779–783PubMed

10.

Siegel JP, Puri RK (1991) Interleukin-2 toxicity. J Clin Oncol 9:694–704PubMed

11.

Maas RA, Dullens HF, Den Otter W (1993) Interleukin-2 in cancer treatment: disappointing or (still) promising? A review. Cancer Immunol Immunother 36:141–148PubMedCrossRef

12.

Shanafelt AB, Lin Y, Shanafelt MC, Forte CP, Dubois-Stringfellow N, Carter C, Gibbons JA, Cheng SL, Delaria KA, Fleischer R, Greve JM, Gundel R, Harris K, Kelly R, Koh B, Li Y, Lantz L, Mak P, Neyer L, Plym MJ, Roczniak S, Serban D, Thrift J, Tsuchiyama L, Wetzel M, Wong M, Zolotorev A (2000) A T-cell-selective interleukin-2 mutein exhibits potent antitumor activity and is well tolerated in vivo. Nat Biotechnol 6:1197–1202CrossRef

13.

Ju G, Collins L, Kaffka KL, Tsien W, Simpson R (1987) Structure-function analysis of human interleukin-2. Identification of amino acid residues required for biological activity. J Biol Chem 262:5723–5731PubMed

14.

Smith KA (1988) Interleukin-2: inception, impact, and implications. Science 240:1169–1176PubMedCrossRef

15.

Moya G, González LJ, Huerta V, García Y, Morera V, Pérez D, Breña F, Araña M (2002) Isolation and characterization of modified species of a mutated (Cys125-Ala) recombinant human interleukin-2. J Chromatogr A 971:129–142PubMedCrossRef

16.

Liu M, Wang B, Sun G, Qian D, Yan Z, Song X, Ding S (2010) Expression, purification, and characterization of a functional mutant recombinant human interleukin-2. Protein Pept Lett 17:1280–1284PubMedCrossRef

17.

Zhu X, Wang J, Wang R, Zhang X (2000) Immunomodulatory effects of bee venom on S180 sarcoma mice. Pharmacol Clin Chin Materia Medica 16:24–25

18.

Wang Q, Lu Y, Zang Y, Zhang Y (2000) Study on the immune-regulating mechanism of the bee venom. Chin J Immunol 16:542–544

19.

Liu L, Ling C, Huang X (2003) Study on purification of melittin and its effect on anti-tumor in vitro. Chin J Biochem Pharm 24:163–166

20.

Jo M, Park MH, Kollipara PS, An BJ, Song HS, Han SB, Kim JH, Song MJ, Hong JT (2012) 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 258:72–81PubMedCrossRef

21.

Saini SS, Chopra AK, Peterson JW (1999) Melittin activates endogenous phospholipase D during cytolysis of human monocytic leukemia cells. Toxicon 37:1605–1619PubMedCrossRef

22.

Arora AS, de Groen PC, Croall DE, Emori Y, Gores GJ (1996) Hepatocellular carcinoma cells resist necrosis during anoxia by preventing phospholipase-mediated calpain activation. J Cell Physiol 167:434–442PubMedCrossRef

23.

Kubo H, Loegering DA, Adophson CR, Gleich GJ (1999) Cytotoxic properties of eosinophil granule major basic protein for tumor cells. Int Arch Allergy Immunol 118:426–428PubMedCrossRef

24.

Lazarev VN, Parfenova TM, Gularyan SK, Misyurina OY, Akopian TA, Govorun VM (2002) Induced expression of melittin, an antimicrobial peptide, inhibits infection by Chlamydia trachomatis and Mycoplasma hominis in a Hela cell line. Int J Antimicrob Agents 19:133–137PubMedCrossRef

25.

Tarhini AA, Agarwala SS (2005) Interleukin-2 for the treatment of melanoma. Curr Opin Investig Drugs 6:1234–1239PubMed

26.

Wang C, Chen T, Zhang N, Yang M, Li B, Lü X, Cao X, Ling C (2009) 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-NFκB. J Biol Chem 284:3804–3813PubMedCrossRef

27.

Park JH, Jeong YJ, Park KK, Cho HJ, Chung IK, Min KS, Kim M, Lee KG, Yeo JH, Park KK, Chang YC (2010) Melittin suppresses PMA-induced tumor cell invasion by inhibiting NF-kappaB and AP-1-dependent MMP-9 expression. Mol Cells 29:209–215PubMedCrossRef

28.

Park MH, Choi MS, Kwak DH, Oh KW, Yoon DY, Han SB, Song HS, Song MJ, Hong JT (2011) Anti-cancer effect of bee venom in prostate cancer cells through activation of caspase pathway via inactivation of NF-ΚB. Prostate 71:801–812PubMedCrossRef

29.

Liu S, Yu M, He Y, Xiao L, Wang F, Song C, Sun S, Ling C, Xu Z (2008) Melittin prevents liver cancer cell metastasis through inhibition of the Rac1-dependent pathway. Hepatology 47:1964–1973PubMedCrossRef

30.

Holle L, Song W, Holle E, Wei Y, Wagner T, Yu X (2003) A matrix metalloproteinase 2 cleavable melittin/avidin conjugate specifically targets tumor cells in vitro and in vivo. Int J Oncol 22:93–98PubMed

31.

Belmont HJ, Price-Schiavi S, Liu B, Card KF, Lee HI, Han KP, Wen J, Tang S, Zhu X, Merrill J, Chavillaz PA, Wong JL, Rhode PR, Wong HC (2006) Potent antitumor activity of a tumor-specific soluble TCR/IL-2 fusion protein. Clin Immunol 121:29–39PubMedCrossRef

32.

Penafuerte C, Bautista-Lopez N, Boulassel MR, Routy JP, Galipeau J (2009) The human ortholog of granulocyte macrophage colony-stimulating factor and interleukin-2 fusion protein induces potent ex vivo natural killer cell activation and maturation. Cancer Res 69:9020–9028PubMedCrossRef

33.

Rosenberg SA, Lotze MT, Muul LM, Chang AE, Avis FP, Leitman S, Linehan WM, Robertson CN, Lee RE, Rubin JT, Seipp CA, Simpson CG, White DE (1987) A progress report on the treatment of 157 patients with advanced cancer using lymphokine-activated killer cells and interleukin-2 or high-dose interleukin-2 alone. N Engl J Med 316:889–897PubMedCrossRef

34.

Grimm EA, Mazumder A, Zhang HZ, Rosenberg SA (1982) Lymphokine-activated killer cell phenomenon. Lysis of natural killer-resistant fresh solid tumor cells by interleukin 2-activated autologous human peripheral blood lymphocytes. J Exp Med 155:1823–1841PubMedCrossRef

Title: A novel melittin-MhIL-2 fusion protein inhibits the growth of human ovarian cancer SKOV3 cells in vitro and in vivo tumor growth
Authors: Mingjun Liu
Jinbao Zong
Zimin Liu
Ling Li
Xu Zheng
Bin Wang
Guirong Sun
Publication date: 01-05-2013
Publisher: Springer-Verlag
Published in: Cancer Immunology, Immunotherapy / Issue 5/2013
Print ISSN: 0340-7004
Electronic ISSN: 1432-0851
DOI: https://doi.org/10.1007/s00262-013-1401-2

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