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Journal of Experimental & Clinical Cancer Research

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Open Access 01-12-2017 | Research

Combination therapy of PKCζ and COX-2 inhibitors synergistically suppress melanoma metastasis

Authors: Ping Zhou, Jiaqi Qin, Yuan Li, Guoxia Li, Yinsong Wang, Ning Zhang, Peng Chen, Chunyu Li

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2017

Abstract

Background

Metastatic malignant melanoma is one of the most aggressive malignancies and its treatment remains challenging. Recent studies demonstrate that the melanoma metastasis has correlations with the heightened activations of protein kinase C ζ (PKCζ) and cyclooxygenase-2 (COX-2) signaling pathways. Targeted inhibitions for PKCζ and COX-2 have been considered as the promising strategies for the treatment of melanoma metastasis. Thus, the PKCζ inhibitor J-4 and COX-2 inhibitor Celecoxib were combined to treat melanoma metastasis in this study.

Methods

The Transwell assay, Wound-healing assay and Adhesion assay were used to evaluate the inhibition of combined therapy of J-4 and Celecoxib on melanoma cells invasion, migration and adhesion in vitro, respectively. The impaired actin polymerization was observed by confocal microscope and inactivated signal pathways about PKCζ and COX-2 were confirmed by the Western blotting assay. The B16-F10/C57BL mouse melanoma model was used to test the inhibition of combined therapy of J-4 and Celecoxib on melanoma metastasis in vivo.

Results

The in vitro results showed that the combination of J-4 and Celecoxib exerted synergistic inhibitory effects on the migration, invasion and adhesion of melanoma B16-F10 and A375 cells with combination index less than 1. The actin polymerization and phosphorylation of Cofilin required in cell migration were severely impaired, which is due to the inactivation of PKCζ related signal pathways and the decrease of COX-2. The combined inhibition of PKCζ and COX-2 induced Mesenchymal-Epithelial Transition (MET) in melanoma cells with the expression of E-Cadherin increasing and Vimentin decreasing. The secretion of MMP-2/MMP-9 also significantly decreased after the combination treatment. In C57BL/6 mice intravenously injected with B16-F10 cells (5 × 10⁴ cells/mouse), co-treatment of J-4 and Celecoxib also severely suppressed melanoma lung metastasis. The body weight monitoring and HE staining results indicated the low toxicity of the combination therapy.

Conclusions

This study demonstrates that the combination therapy of PKCζ and COX-2 inhibitors can significantly inhibit melanoma metastasis in vitro and in vivo, which will be an efficient strategy for treatment of melanoma metastasis in clinics.

Dunn J, Watson M. Aitken JF. Systematic Review of Psychosocial Outcomes for Patients with Advanced Melanoma. Psychooncology: Hyde MK; 2016. in press

Park SM, Li T, Wu S, Li WQ, Weinstock M, Qureshi AA, Cho E. Niacin intake and risk of skin cancer in US women and men. Int J Cancer. 2017;140:2023–31.CrossRefPubMed

Loquai C, Schmidtmann I, Garzarolli M, Kaatz M, Kahler KC, Kurschat P, Meiss F, Micke O, Muecke R, Muenstedt K, et al. Interactions from complementary and alternative medicine in patients with melanoma. Melanoma Res. 2017; in press

Chhabra A, Mukherji B, Batra D. Activation induced cell death (AICD) of human melanoma antigen-specific TCR engineered CD8 T cells involves JNK, Bim and p53. Expert Opin Ther Targets. 2017;21:117–29.CrossRefPubMed

Polkowska M, Czepielewska E, Kozlowska-Wojciechowska M. Drug combinations as the new standard for melanoma treatment. Curr Treat Options in Oncol. 2016;17:61.CrossRef

Madeddu C, Dessi M, Panzone F, Serpe R, Antoni G, Cau MC, Montaldo L, Mela Q, Mura M, Astara G, et al. Randomized phase III clinical trial of a combined treatment with carnitine plus celecoxib +/− megestrol acetate for patients with cancer-related anorexia/cachexia syndrome. Clin Nutr. 2012;31:176–82.CrossRefPubMed

Li Z, Ye S, OuYang L, Zhang H, Chen Y, He J, Chen Q, Qian C, Zhang X, Cui J, et al: COX-2 promotes metastasis in nasopharyngeal carcinoma by mediating interactions between cancer cells and myeloid-derived suppressor cells. Oncoimmunology 2015, 4:e1044712. eCollection.

Guo Z, Jiang J, Zhang J, Yang H, Yang F, Qi Y, Zhong Y, Su J, Yang R, Li L, Xiang B. COX-2 promotes migration and invasion by the side population of cancer stem cell-like hepatocellular carcinoma cells. Medicine. 2015;94:e1806.CrossRefPubMedPubMedCentral

Cao C, Gao R, Zhang M, Amelio AL, Fallahi M, Chen Z, Gu Y, Hu C, Welsh EA, Engel BE, et al. Role of LKB1-CRTC1 on glycosylated COX-2 and response to COX-2 inhibition in lung cancer. J Natl Cancer Inst. 2015;107:358.CrossRefPubMed

10.

Kim KM, Im A, Kim SH, Hyun JW, Chae S. Timosaponin AIII inhibits melanoma cell migration by suppressing COX-2 and in vivo tumor metastasis. Cancer Sci. 2016;107:181–8.CrossRefPubMedPubMedCentral

11.

Goulet AC, Einsphar JG, Alberts DS, Beas A, Burk C, Bhattacharyya A, Bangert J, Harmon JM, Fujiwara H, Koki A, Nelson MA. Analysis of cyclooxygenase 2 (COX-2) expression during malignant melanoma progression. Cancer Biol Ther. 2003;2:713–8.CrossRefPubMed

12.

Panza E, De Cicco P, Ercolano G, Armogida C, Scognamiglio G, Anniciello AM, Botti G, Cirino G, Ianaro A. Differential expression of cyclooxygenase-2 in metastatic melanoma affects progression free survival. Oncotarget. 2016;7:57077–85.CrossRefPubMedPubMedCentral

13.

Szymanski PT, Muley P, Ahmed SA, Khalifa S, Fahmy H. Sarcophine-diol inhibits expression of COX-2, inhibits activity of cPLA2, enhances degradation of PLA2 and PLC(gamma)1 and inhibits cell membrane permeability in mouse melanoma B16F10 cells. Mar Drugs. 2012;10:2166–80.CrossRefPubMedPubMedCentral

14.

Adler NR, Haydon A, McLean CA, Kelly JW, Mar VJ. Metastatic pathways in patients with cutaneous melanoma. Pigment Cell Melanoma Res. 2017;30:13–27.CrossRefPubMed

15.

El-Rayes BF, Zalupski MM, Shields AF, Ferris AM, Vaishampayan U, Heilbrun LK, Venkatramanamoorthy R, Adsay V, Philip PA. A phase II study of celecoxib, gemcitabine, and cisplatin in advanced pancreatic cancer. Invest New Drug. 2005;23:583–90.CrossRef

16.

Elmets CA, Viner JL, Pentland AP, Cantrell W, Lin H, Bailey H, Kang S, Linden KG, Heffernan M, Duvic M, et al. Chemoprevention of nonmelanoma skin cancer with celecoxib: a randomized, double-blind, placebo-controlled trial. J Natl Cancer Inst. 2010;102:1835–44.CrossRefPubMedPubMedCentral

17.

Javle MM, Cao S, Durrani FA, Pendyala L, Lawrence DD, Smith PF, Creaven PJ, Noel DC, Iyer RV, Rustum YM. Celecoxib and mucosal protection: translation from an animal model to a phase I clinical trial of celecoxib, irinotecan, and 5-fluorouracil. Clin Cancer Res. 2007;13:965–71.CrossRefPubMed

18.

Seo KW, Coh YR, Rebhun RB, Ahn JO, Han SM, Lee HW, Youn HY. Antitumor effects of celecoxib in COX-2 expressing and non-expressing canine melanoma cell lines. Res Vet Sci. 2014;96:482–6.CrossRefPubMedPubMedCentral

19.

Sadhu SS, Wang S, Averineni RK, Seefeldt T, Yang Y, Guan X. In-vitro and in-vivo inhibition of melanoma growth and metastasis by the drug combination of celecoxib and dacarbazine. Melanoma Res. 2016;26:572–9.CrossRefPubMed

20.

Gowda R, Sharma A, Robertson GP. Synergistic inhibitory effects of celecoxib and Plumbagin on melanoma tumor growth. Cancer Lett. 2017;385:243–50.CrossRefPubMed

21.

Niessner H, Sinnberg T, Kosnopfel C, Smalley KSM, Beck D, Praetorius C, Mai M, Beissert S, Kulms D, Schaller M, et al. BRAF inhibitors amplify the pro-apoptotic activity of MEK inhibitors by inducing ER stress in NRAS-mutant melanoma. Clin Cancer Res. 2017; https://doi.org/10.1158/1078-0432.CCR-17-0098. in press. in press.

22.

Escuin-Ordinas H, Atefi M, Fu Y, Cass A, Ng C, Huang RR, Yashar S, Comin-Anduix B, Avramis E, Cochran AJ, et al. COX-2 inhibition prevents the appearance of cutaneous squamous cell carcinomas accelerated by BRAF inhibitors. Mol Oncol. 2014;8:250–60.CrossRefPubMed

23.

Guo H, Ma Y, Zhang B, Sun B, Niu R, Ying G, Zhang N. Pivotal advance: PKC zeta is required for migration of macrophages. J Leukoc Biol. 2009;85:911–8.CrossRefPubMed

24.

Sun RH, Gao P, Chen L, Ma DL, Wang JM, Oppenheim JJ, Zhang N. Protein kinase C zeta is required for epidermal growth factor-induced chemotaxis of human breast cancer cells. Cancer Res. 2005;65:1433–41.CrossRefPubMed

25.

Schondorf T, Kurbacher CM, Becker M, Warm M, Kolhagen H, Gohring UJ. Heterogeneity of proteinkinase C activity and PKC-zeta expression in clinical breast carcinomas. Clin Exp Med. 2001;1:1–8.CrossRefPubMed

26.

Guo H, Gu F, Li W, Zhang B, Niu R, Fu L, Zhang N, Ma Y. Reduction of protein kinase C zeta inhibits migration and invasion of human glioblastoma cells. J Neurochem. 2009;109:203–13.CrossRefPubMed

27.

Liu Y, Wang B, Wang J, Wan W, Sun R, Zhao Y, Zhang N. Down-regulation of PKCzeta expression inhibits chemotaxis signal transduction in human lung cancer cells. Lung Cancer. 2009;63:210–8.CrossRefPubMed

28.

Krasagakis K, Fimmel S, Genten D, Eberle J, Quas P, Ziegler W, Haller H, Orfanos CE. Lack of protein kinase C (PKC)-beta and low PKC-alpha, −delta, −epsilon, and -zeta isozyme levels in proliferating human melanoma cells. Int J Oncol. 2002;20:865–71.PubMed

29.

Certa U, Seiler M, Padovan E, Spagnoli GC. Interferon-a sensitivity in melanoma cells: detection of potential response marker genes. Recent Results Cancer Res. 2002;160:85–91.CrossRefPubMed

30.

Oka M, Kikkawa U. Protein kinase C in melanoma. Cancer Metastasis Rev. 2005;24:287–300.CrossRefPubMed

31.

Tsubaki M, Matsuoka H, Yamamoto C, Kato C, Ogaki M, Satou T, Itoh T, Kusunoki T, Tanimori Y, Nishida S. The protein kinase C inhibitor, H7, inhibits tumor cell invasion and metastasis in mouse melanoma via suppression of ERK1/2. Clin Exp Metastasis. 2007;24:431–8.CrossRefPubMed

32.

Hodgson L, Henderson AJ, Dong C. Melanoma cell migration to type IV collagen requires activation of NF-kappaB. Oncogene. 2003;22:98–108.CrossRefPubMedPubMedCentral

33.

Wu J, Liu S, Fan Z, Zhang L, Tian Y, Yang R. A novel and selective inhibitor of PKC zeta potently inhibits human breast cancer metastasis in vitro and in mice. Tumor Biol. 2016;37:8391–401.CrossRef

34.

Wu J, Zhang B, Wu M, Li H, Niu R, Ying G, Zhang N. Screening of a PKC zeta-specific kinase inhibitor PKCzI257.3 which inhibits EGF-induced breast cancer cell chemotaxis. Invest New Drug. 2010;28:268–75.CrossRef

35.

Li H, Wu J, Ying G, Chen L, Lai L, Liu Z, Zhang N, Guo H. J-4: a novel and typical preclinical anticancer drug targeting protein kinase C zeta. Anti-Cancer Drug. 2012;23:691–7.CrossRef

36.

Lu S, Smith AP, Moore D, Lee NM. Different real-time PCR systems yield different gene expression values. Mol Cell Probes. 2010;24:315–20.CrossRefPubMed

37.

Duary RK, Bhausaheb MA, Batish VK, Grover S. Anti-inflammatory and immunomodulatory efficacy of indigenous probiotic lactobacillus plantarum Lp91 in colitis mouse model. Mol Biol Rep. 2012;39:4765–75.CrossRefPubMed

38.

Chang MS, Choi MJ, Park SY, Park SK. Inhibitory effects of Hoelen extract on Melanogenesis in B16/F1 melanoma cells. Phytother Res. 2010;24:1359–64.CrossRefPubMed

39.

Zhang N, Fu J, Chou T. Synergistic combination of microtubule targeting anticancer fludelone with cytoprotective panaxytriol derived from panax ginseng against MX-1 cells in vitro: experimental design and data analysis using the combination index method. Am J Cancer Res. 2016;6:97–104.PubMed

40.

Hussain S, Assender JW, Bond M, Wong LF, Murphy D, Newby AC. Activation of protein kinase Czeta is essential for cytokine-induced metalloproteinase-1, −3, and −9 secretion from rabbit smooth muscle cells and inhibits proliferation. J Biol Chem. 2002;277:27345–52.CrossRefPubMed

41.

Goichberg P, Kalinkovich A, Borodovsky N, Tesio M, Petit I, Nagler A, Hardan I, Lapidot T. cAMP-induced PKCzeta activation increases functional CXCR4 expression on human CD34+ hematopoietic progenitors. Blood. 2006;107:870–9.CrossRefPubMed

42.

Wang Z, Chen J, Wang J, Ahn S, Li C, Lu Y, Loveless VS, Dalton JT, Miller DD, Li W. Novel tubulin polymerization inhibitors overcome multidrug resistance and reduce melanoma lung metastasis. Pharm Res-Dordr. 2012;29:3040–52.CrossRef

43.

Amarzguioui M, Peng Q, Wiiger MT, Vasovic V, Babaie E, Holen T, Nesland JM, Prydz H. Ex vivo and in vivo delivery of anti-tissue factor short interfering RNA inhibits mouse pulmonary metastasis of B16 melanoma cells. Clin Cancer Res. 2006;12:4055–61.CrossRefPubMed

44.

Bell E, Ponthan F, Whitworth C, Tweddle DA, Lunec J, Redfern CPF. COX2 expression in neuroblastoma increases tumorigenicity but does not affect cell death in response to the COX2 inhibitor celecoxib. Clin Exp Metastas. 2014;31:651–9.CrossRef

45.

Kim SH, Kim Y, Kim M, Kim DS, Lee SC, Chi S, Lee DH, Park SG, Park BC, Bae K, Kang S. Comparative proteomic analysis of mouse melanoma cell line B16, a metastatic descendant B16F10, and B16 overexpressing the metastasis-associated tyrosine phosphatase PRL-3. Oncol Res. 2009;17:601–12.CrossRefPubMed

Title: Combination therapy of PKCζ and COX-2 inhibitors synergistically suppress melanoma metastasis
Authors: Ping Zhou
Jiaqi Qin
Yuan Li
Guoxia Li
Yinsong Wang
Ning Zhang
Peng Chen
Chunyu Li
Publication date: 01-12-2017
Publisher: BioMed Central
Published in: Journal of Experimental & Clinical Cancer Research / Issue 1/2017
Electronic ISSN: 1756-9966
DOI: https://doi.org/10.1186/s13046-017-0585-2

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