The experimental study of establishing tumor-associated macrophage model

doi:10.3969/j.issn.1673-8640.2014.09.007

Abstract

Abstract:

Objective To establish the differentiation model of tumor-associated macrophages (TAM) in vitro. Methods Monocytes were isolated from the peripheral blood of healthy subjects and cultured in vitro. After stimulation by interleukin 4(IL-4) and interleukin 13(IL-13), the expression changes of molecule markers on monocytes were determined by flow cytometry, including CD14, CD204 and CD206. The levels of interleukin 10 (IL-10) secreted by monocytes weredetermined by enzyme-linked iummunosorbent assay (ELISA). After coculturing with monocytes, the apoptosis of tumor cells was investigated by Hoechst staining. Results After stimulation by IL-4 and IL-13, monocytes exhibited a markedly altered phenotype with retained CD14, the increasing expressions of CD204 and CD206 and the increasing amount of secreted IL-10. Moreover, when coculturing with tumor cells, the cytotoxity of monocytes on tumor cells was inhibited by IL-4/IL-13. Conclusions IL-4/IL-13 could induce monocytes to exhibit TAM phenotype and function, indicating IL-4/IL-13 could be applied to the establishment of TAM differentiation model, which can be used inthe research of TAM.

Key words: Tumor-associated macrophage, Interleukin 4, Interleukin 13, Monocyte, M2 macrophage

CLC Number:

R446.61

ZHANG Guoliang, LIU Yiwen, HE Yiqing, YANG Cuixia, WANG Wenjuan, DU Yan, GAO Feng. The experimental study of establishing tumor-associated macrophage model[J]. , 2014, 29(9): 897-802.

References

[1] Mantovani A, Sozzani S, Locati M, et al. Macrophage polarization: tumor-associated macrophages as a paradigm for polarized M2 mononuclear phagocytes[J]. Trends Immunol, 2002, 23(11):549-555.
[2] Leek RD, Lewis CE, Whitehouse R, et al. Association of macrophage infiltration with angiogenesis and prognosis in invasive breast carcinoma[J]. Cancer Res, 1996, 56(20):4625-4629.
[3] Nowicki A, Szenajch J, Ostrowska G, et al. Impaired tumor growth in colony-stimulating factor 1 (CSF-1)-deficient, macrophage-deficient op/op mouse: evidence for a role of CSF-1-dependent macrophages in formation of tumor stroma[J]. Int J Cancer, 1996, 65(1):112-119.
[4] Priceman SJ, Sung JL, Shaposhnik Z, et al. Targeting distinct tumor-infiltrating myeloid cells by inhibiting CSF-1 receptor: combating tumor evasion of antiangiogenic therapy[J]. Blood, 2010, 115(7):1461-1471.
[5] Tjiu JW, Chen JS, Shun CT, et al. Tumor-associated macrophage-induced invasion and angiogenesis of human basal cell carcinoma cells by cyclooxygenase-2 induction[J]. J Invest Dermatol, 2009, 129(4): 1016-1025.
[6] Byrne SN, Knox MC, Halliday GM. TGFbeta is responsible for skin tumour infiltration by macrophages enabling the tumours to escape immune destruction[J]. Immunol Cell Biol, 2008, 86(1):92-97.
[7] Zhou W, Bao S. Reciprocal supportive interplay between glioblastoma and tumor-associated macrophages[J].Cancers (Basel), 2014, 6(2):723-740.
[8] Hagemann T, Robinson SC, Schulz M, et al. Enhanced invasiveness of breast cancer cell lines upon co-cultivation with macrophages is due to TNF-alpha dependent up-regulation of matrix metalloproteases[J]. Carcinogenesis, 2004, 25(8):1543-1549.
[9] Goswami S, Sahai E, Wyckoff JB, et al. Macrophages promote the invasion of breast carcinoma cells via a colony-stimulating factor-1/epidermal growth factor paracrine loop[J]. Cancer Res, 2005, 65(12):5278-5283.
[10] Toge H, Inagaki T, Kojimoto Y, et al. Angiogenesis in renal cell carcinoma: the role of tumor-associated macrophages[J]. Int J Urol, 2009, 16(10):801-807.
[11] Riabov V, Gudima A, Wang N, et al. Role of tumor associated macrophages in tumor angiogenesis and lymphangiogenesis[J]. Front Physiol, 2014, 5:75.
[12] Jeon BH, Jang C, Han J, et al. Profound but dysfunctional lymphangiogenesis via vascular endothelial growth factor ligands from CD11b+ macrophages in advanced ovarian cancer[J]. Cancer Res, 2008, 68(4):1100-1109.
[13] Shree T, Olson OC, Elie BT, et al. Macrophages and cathepsin proteases blunt chemotherapeutic response in breast cancer[J]. Genes Dev, 2011, 25(23): 2465-2479.
[14] Jinushi M, Chiba S, Yoshiyama H, et al. Tumor-associated macrophages regulate tumorigenicity and anticancer drug responses of cancer stem/initiating cells[J]. Proc Natl Acad Sci USA, 2011, 108(30): 12425-12430.
[15] Freytes DO, Kang JW, Marcos-Campos I, et al. Macrophages modulate the viability and growth of human mesenchymal stem cells[J]. J Cell Biochem, 2013, 114(1):220-229.
[16] Tjiu JW, Chen JS, Shun CT, et al. Tumor-associated macrophage-induced invasion and angiogenesis of human basal cell carcinoma cells by cyclooxygenase-2 induction[J]. J Invest Dermatol, 2009, 129(4):1016-1025.
[17] Ruffell B, Affara NI, Coussens LM. Differential macrophage programming in the tumor microenvironment[J]. Trends Immunol, 2012, 33(3):119-126.
[18] Werno C, Menrad H, Weigert A, et al. Knockout of HIF-1α in tumor-associated macrophages enhances M2 polarization and attenuates their pro-angiogenic responses[J]. Carcinogenesis, 2010, 31(10):1863-1872.
[19] Komohara Y, Ohnishi K, Kuratsu J, et al. Possible involvement of the M2 anti-inflammatory macrophage phenotype in growth of human gliomas[J]. J Pathol, 2008, 216(1):15-24.