Enhanced metastasis in RNF13 knockout mice is mediated by a reduction in GM-CSF levels

Abstract

RING finger protein 13 (RNF13) is a novel E3 ubiquitin ligase whose expression is associated with cancer development. However, its specific role in cancer progression and metastasis remains unclear. Here, a B16F10/LLC experimental pulmonary metastatic model was developed to examine the formation of metastatic foci in the lung. A greater number of tumor colonies were observed in the lungs of RNF13-knockout (KO) mice than in their wild-type (WT) littermates, whereas no significant differences in tumor size were observed between the two groups. In short-term experiments, the number of fluorescently-labeled B16F10 cells increased remarkably in RNF13-KO lungs at early time points, whereas clearance of tumor cells from the blood was not affected. These results indicated that RNF13 may inhibit the colonization of B16F10 cells in the lung. Assessment of the concentration of various cytokines in tumor bearing lungs and blood did not detect significant differences between the blood of RNF13-KO and WT mice; however the levels of GM-CSF were significantly reduced in RNF13-KO tumor bearing lungs, which may have guided more B16F10 cells to migrate to the lungs. This was confirmed by lower GM-CSF concentrations in conditioned media from the culture of RNF13-KO lung slices. Collectively, our results suggest that host RNF13 affects the concentration of GM-CSF in tumor-bearing lungs, leading to a reduction in the colonization of metastatic tumor cells in the lung.

Keywords

RNF13 / metastasis / GM-CSF / lung cancer

References

[1] Bayne LJ, Beatty GL, Jhala N, Clark CE, Rhim AD, Stanger BZ, Vonderheide RH (2012) Tumor-derived granulocyte-macrophage colony-stimulating factor regulates myeloid inflammation and T cell immunity in pancreatic cancer. Cancer Cell 21: 822−835

[2] Bocock JP, Carmicle S, Chhotani S, Ruffolo MR, Chu H, Erickson AH (2009) The PA-TM-RING protein RING finger protein 13 is an endosomal integral membrane E3 ubiquitin ligase whose RING finger domain is released to the cytoplasm by proteolysis. FEBS J 276: 1860−1877

[3] Dranoff G (2002) GM-CSF-based cancer vaccines. Immunol Rev 188: 147−154

[4] Dranoff G (2003) GM-CSF-secreting melanoma vaccines. Oncogene 22: 3188−3192

[5] Eubank TD, Roberts RD, Khan M, Curry JM, Nuovo GJ, Kuppusamy P, Marsh CB (2009) Granulocyte macrophage colony-stimulating factor inhibits breast cancer growth and metastasis by invoking an anti-angiogenic program in tumor-educated macrophages. Cancer Res 69: 2133−2140

[6] Fidler IJ (1970) Metastasis: guantitative analysis of distribution and fate of tumor embolilabeled with 125 I-5-iodo-2′-deoxyuridine. J Natl Cancer Inst 45: 773−782

[7] Hanahan D, Weinberg RA (2011) Hallmarks of cancer: the next generation. Cell 144: 646−674

[8] Irmisch A, Huelsken J (2013) Metastasis: new insights into organspecific extravasation and metastatic niches. Exp cell Res 319: 1604−1610

[9] Jin X, Cheng H, Chen J, Zhu D (2011) RNF13: an emerging RING finger ubiquitin ligase important in cell proliferation. FEBS J 278: 78−84

[10] Joyce JA, Pollard JW (2009) Microenvironmental regulation of metastasis. Nat Rev Cancer 9: 239−252

[11] Luzzi KJ, MacDonald IC, Schmidt EE, Kerkvliet N, Morris VL, Chambers AF, Groom AC (1998) Multistep nature of metastatic inefficiency: dormancy of solitary cells after successful extravasation and limited survival of early micrometastases. Am J Pathol 153: 865−873

[12] Man YG, Mason J, Harley R, Kim YH, Zhu K, Gardner WA (2011) Leukocyte-mediated cell dissemination and metastasis: findings frommultiple typesofhuman tumors. JCell Biochem1 12: 1154−1167

[13] Meng J, Zou X, Wu R, Zhong R, Zhu D, Zhang Y (2014) Accelerated regeneration of the skeletal muscle in RNF13-knockout mice is mediated by macrophage-secreted IL-4/IL-6. Protein Cell 5: 235−247

[14] Nguyen DX, Bos PD, Massague J (2009) Metastasis: from dissemination to organ-specific colonization. Nat Rev Cancer 9: 274−284

[15] Nicolson GL (1979) Cancer metastasis. Sci Am 240: 66−76

[16] Paget S (1989) The distribution of secondary growths in cancer of the breast. 1889. Cancer Metastasis Rev 8: 98−101

[17] Spano D, Zollo M (2012) Tumor microenvironment: a main actor in the metastasis process. Clin Exp Metastasis 29: 381−395

[18] Spitler, L.E., Cao, H., Piironen, T., Whiteside, T.L., Weber, R.W., and Cruickshank, S. (2014). Biological effects of anti-granulocytemacrophage colony-stimulating factor (GM-CSF) antibody formation in patients treated WithGM-CSF(Sargramostim) as adjuvant therapy of melanoma. Am J Clin Oncol.

[19] Stanley AC, Lacy P (2010) Pathways for cytokine secretion. Physiol (Bethesda) 25: 218−229

[20] Talmadge JE, Fidler IJ (2010) AACR centennial series: the biology of cancer metastasis: historical perspective. Cancer Res 70: 5649−5669

[21] Tranque P, Crossin KL, Cirelli C, Edelman GM, Mauro VP (1996) Identification and characterization of a RING zinc finger gene (CRZF) expressed in chicken embryo cells. Proc Natl Acad Sci USA 93: 3105−3109

[22] van Dijk JR, Yamazaki Y, Palmer RH (2014) Tumour-associated mutations of PA-TM-RING ubiquitin ligases RNF167/RNF13 identify the PA domain as a determinant for endosomal localization. Biochem J 459: 27−36

[23] Williams P, Bouchentouf M, Rafei M, Romieu-Mourez R, Hsieh J, Boivin MN, Yuan S, Forner KA, Birman E, Galipeau J (2010) A dendritic cell population generated by a fusion of GM-CSF and IL-21 induces tumor-antigen-specific immunity. J Immunol 185: 7358−7366

[24] Zhang Q, Meng Y, Zhang L, Chen J, Zhu D (2009) RNF13: a novel RING-type ubiquitin ligase over-expressed in pancreatic cancer. Cell Res 19: 348−357

[25] Zhang Q, Wang K, Zhang Y, Meng J, Yu F, Chen Y, Zhu D (2010) The myostatin-induced E3 ubiquitin ligase RNF13 negatively regulates the proliferation of chicken myoblasts. FEBS J 277: 466−476

[26] Zhang Q, Li Y, Zhang L, Yang N, Meng J, Zuo P, Zhang Y, Chen J, Wang L, Gao X (2013) E3 ubiquitin ligase RNF13 involves spatial learning and assembly of the SNARE complex. Cell Mol Life Sci 70: 153−165

RIGHTS & PERMISSIONS

2014 This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.