Effects of JNK Pathway on Polarization and Pro-tumor Function of M2 Macrophage

doi:10.13523/j.cb.20180401

China Biotechnology

2018, Vol. 38

Issue (4): 1-7 DOI: 10.13523/j.cb.20180401

Effects of JNK Pathway on Polarization and Pro-tumor Function of M2 Macrophage

Jin HAO¹,Zi-xin ZHU²,Xiao-yan LV³,Qin ZHOU^1*

1 Key Laboratory of Laboratory Medical Diagnostics of Education Ministry,College of Laboratory Medicine, Chongqing Medical University,Chongqing 400016,China
2 Chongqing No.8 Secondary School,Chongqing 400016, China
3 Department of Dermatology, West China Hospital, Chengdu 610000, China

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Abstract

The present study investigates the effect of JNK pathway on the polarization of M2 status as well as pro-tumor function mediated by M2. THP1 derived M2 macrophage (THP1-M2) model was established. The cells were divided into 3 groups: the PMA pretreated unpolarized macrophage (M0), the PMA and IL-4 induced M2 macrophage with DMSO (negative control) treated (M2), the JNK inhibitor treated M2 macrophage (M2-JNK I). Furthermore, the M2 associated markers Arginase1 (Arg1),mannose receptor C-type 1 (Mrc1)were analyzed by Q-PCR, the protein level of Arg1 and Mrc1 were detected by Western blot, the migration ability of macrophages was tested by Wound Healing, the apoptosis of 786O and OSRC2 were analyzed by flow cytometry. Compared with the THP1-M2, THP1-M2-JNK I group showed decreased expression of Arg1 and Mrc1, and migration ability was impaired. What’s more, block of JNK pathway inhibited the pro-tumor function of M2 on 786O and OSCR2. Taken together, the results suggest that inhibition of JNK pathway regulates M2 polarization and its pro-tumor effects.

Key words： JNK pathway M2 polarization Tumor IL-4

Received: 19 October 2017 Published: 08 May 2018

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	Jin HAO
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Cite this article:

Jin HAO,Zi-xin ZHU,Xiao-yan LV,Qin ZHOU. Effects of JNK Pathway on Polarization and Pro-tumor Function of M2 Macrophage. China Biotechnology, 2018, 38(4): 1-7.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20180401 OR https://manu60.magtech.com.cn/biotech/Y2018/V38/I4/1

Table 1 The list of the primers

Fig.1 Expression of M2 associated markers in THP1-M2
(a) THP1-M0 cells were stimulated with IL-4 (20ng/ml) for 12h, 14h, 16h, 20h and 24h. The expression of M2-associated genes, including Arg1, Mrc1 was evaluated by qPCR (b) Stimulated with IL-4 or 5% BSA on M0 for 16h, and the expression of M2 associated cytokine genes Arg1 and Mrc1 were analyzed with qPCR (c) Stimulated with IL-4 on M0 for 16h, Arg1 and Mrc1 protein levels were analyzed with Western blot ^* P<0.05, ^** P<0.01

Fig.2 Effects of JNK I IRF1on the polarization of on THP1-M2
(a) THP1-M0 macrophages were treated with JNK-inhibitor I (JNK-I, 20μmol/L) or DMSO for 60min, followed by stimulation with IL-4 for 16h. Arg1, Mrc1 expression was determined by quantitative PCR (b) Protein levels of Arg1, Mrc1 were evaluated by Western blot. Band intensity of Western blot was analyzed by densitometry, and normalized to β-actin level ^* P<0.05, ^** P<0.01

Fig.3 Silencing of JNK pathway inhibits THP1-M2 cells migration
(a) Cell migration of THP1 cells was measured via wound healing. The width of wound area was calculated at three time points (0h, 12h, 24h) starting from the point when THP-1 cells were treated with IL-4 (b) The quantification of wound healing percentage (100%) was exhibited with the error bars representing mean ± SD of three independent experiments ^* P<0.05, ^** P<0.01

Fig.4 Effects of conditioned medium on apoptosis of renal cell carcinoma analyzed by flow cytometry
(a) The numbers in parentheses represented early apoptotic rate and late apoptotic rate (quadrant 2 plus quadrant 3) (b) Histograms showed apoptotic rate of 786O and OSRC2 cultured with CMs ^* P<0.05, ^**P<0.01


[1]	Sica A, Mantovani A . Macrophage plasticity and polarization: in vivo veritas. Journal of Clinical Investigation, 2006,108(2):408-409.

[2]	Chinetti-Gbaguidi G, Colin S, Staels B . Macrophage subsets in atherosclerosis. Nature Reviews Cardiology, 2015,12(1):10. doi: 10.1038/nrcardio.2014.173

[3]	Wynn T A, Chawla A, Pollard J W . Macrophage biology in development, homeostasis and disease. Nature, 2013,496(7446):445-455. doi: 10.1038/nature12034 pmid: 23619691

[4]	Zheng X F, Hong Y X, Feng G J , et al. Lipopolysaccharide-induced M2 to M1 macrophage transformation for IL-12p70 production is blocked by Candida albicans mediated up-regulation of EBI3 expression. PLoS One, 2013,8(5):e63967. doi: 10.1371/journal.pone.0063967 pmid: 3664618

[5]	Martinez F O, Helming L, Gordon S . Alternative activation of macrophages: an immunologic functional perspective. Annual Review of Immunology, 2009,27(1):451-483. doi: 10.1146/annurev.immunol.021908.132532 pmid: 19105661

[6]	Murray P J, Wynn T A . Protective and pathogenic functions of macrophage subsets. Nature Reviews Immunology, 2011,11(11):723-737. doi: 10.1038/nri3073 pmid: 21997792

[7]	Moreno J L, Kaczmarek M, Keegan A D , et al. IL-4 suppresses osteoclast development and mature osteoclast function by a STAT6-dependent mechanism: irreversible inhibition of the differentiation program activated by RANKL. Blood, 2003,102(3):1078-1086. doi: 10.1182/blood-2002-11-3437 pmid: 12689929

[8]	Jiang H, Harris M B, Rothman P . IL-4/IL-13 signaling beyond JAK/STAT. Journal of Allergy and Clinical Immunology, 2000,105(6):1063-1070. doi: 10.1067/mai.2000.107604

[9]	Hao J, Hu Y, Li Y , et al. Involvement of JNK signaling in IL-4 induced M2 macrophage polarization. Experimental Cell Research, 2017,357(2):155-162. doi: 10.1016/j.yexcr.2017.05.010 pmid: 28501460

[10]	Zhou Y, Zhang T, Wang X , et al. Curcumin modulates macrophage polarization through the inhibition of the toll-like receptor 4 expression and its signaling pathways. Cellular Physiology and Biochemistry: International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology, 2015,36(2):631-641.

[11]	Martinez F O, Helming L, Milde R , et al. Genetic programs expressed in resting and IL-4 alternatively activated mouse and human macrophages: similarities and differences. Blood, 2013,121(9):e57-69. doi: 10.1182/blood-2012-06-436212 pmid: 23293084

[12]	Zou J, Shankar N . Roles of TLR/MyD88/MAPK/NF-kappaB signaling pathways in the regulation of phagocytosis and proinflammatory cytokine expression in response to E. faecalis infection. PLoS One, 2015,10(8):e0136947. doi: 10.1371/journal.pone.0136947 pmid: 4552673

[13]	Xiong C Y, Guan D W, Liu Z H , et al. Changes of phospho-JNK expression during the skin burned wound healing. Journal of Forensic Medicine, 2008,24(5):325-335. pmid: 18979912

[14]	Gnanaprakasam J N, Estrada-Muniz E, Vega L . The anacardic 6-pentadecyl salicylic acid induces macrophage activation via the phosphorylation of ERK1/2, JNK, P38 kinases and NF-kappaB. International Immunopharmacology, 2015,29(2):808-817. doi: 10.1016/j.intimp.2015.08.038 pmid: 26371858

[15]	Sica A, Bronte V . Altered macrophage differentiation and immune dysfunction in tumor development. Journal of Clinical Investigation, 2007,117(5):1155-1166. doi: 10.1172/JCI31422 pmid: 17476345

[16]	Nakanishi Y, Nakatsuji M, Seno H , et al. COX-2 inhibition alters the phenotype of tumor-associated macrophages from M2 to M1 in ApcMin/+ mouse polyps. Carcinogenesis, 2011,32(9):1333-1339. doi: 10.1093/carcin/bgr128 pmid: 21730361

[17]	Swartz M A, Iida N, Roberts E W , et al. Tumor microenvironment complexity: emerging roles in cancer therapy. Cancer Research, 2012,72(10):2473. doi: 10.1158/0008-5472.CAN-12-0122 pmid: 22414581

[18]	Smit-Peter S Y . The role of cytokine in regulation of the natural killer cell activity. Srp Arh Celok Lek, 2008,136(7):423-429. doi: 10.2298/SARH0808423J pmid: 18959181

[19]	Engel M A, Neurath M F . Anticancer properties of the IL-12 family--focus on colorectal cancer. Current Medicinal Chemistry, 2010,17(29):3303. doi: 10.2174/092986710793176366 pmid: 20712574

[20]	Hanlon A M, Jang S, Salgame P . Signaling from cytokine receptors that affect Th1 responses. Frontiers in Bioscience A Journal & Virtual Library, 2002,7(7):d1247. doi: 10.2741/hanlon pmid: 11991837

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