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Mast cells promote melanoma colonization of lungs

Öhrvik, Helena, Grujic, Mirjana, Waern, Ida, Gustafson, Ann-Marie, Ernst, Nancy, Roers, Axel, Hartmann, Karin, Pejler, Gunnar·2016·Oncotarget
DOI 10.18632/oncotarget.11837PMC PMC5356606
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Figure 1 Mast cells do not influence the growth of subcutaneous melanoma tumors Melanoma cells (B16.F10) were injected subcutaneously into the flanks of either Mcpt5-Cre− or Mcpt5-Cre+ mice. After 1 or 2 weeks, tumors were excised. A. Staining of subcutaneous tissue of Mcpt5-Cre− or Mcpt5-Cre+ mice with either chloroacetate esterase (left panels) or toluidine blue (right panels). Note the presence of chloroacetate esterase- and toluidine blue-positive mast cells (marked with arrows) in tissue from Mcpt5-Cre− animals (upper panels) and the absence of mast cells in tissue from Mcpt5-Cre+ mice (lower panels). Note also that mast cells are abundant at a distance of ∼300 μm from the tumor border (arrowhead), but not detectable within the tumor mass. Original magnification = 200 x; Bar = 100 μm. B. Representative images of tumors excised from Mcpt5-Cre− and Mcpt5-Cre+ mice 2 weeks after injection of 0.5 − 106 B16.F10 cells. Bar = 5 mm C. Tumor end weights 1 or 2 weeks after injection of 0.5 − 106 B16.F10 cells. Results are shown as mean ± SD; n = 4-7.
Figure 1 Mast cells do not influence the growth of subcutaneous melanoma tumors Melanoma cells (B16.F10) were injected subcutaneously into the flanks of either Mcpt5-Cre− or Mcpt5-Cre+ mice. After 1 or 2 weeks, tumors were excised. A. Staining of subcutaneous tissue of Mcpt5-Cre− or Mcpt5-Cre+ mice with either chloroacetate esterase (left panels) or toluidine blue (right panels). Note the presence of chloroacetate esterase- and toluidine blue-positive mast cells (marked with arrows) in tissue from Mcpt5-Cre− animals (upper panels) and the absence of mast cells in tissue from Mcpt5-Cre+ mice (lower panels). Note also that mast cells are abundant at a distance of ∼300 μm from the tumor border (arrowhead), but not detectable within the tumor mass. Original magnification = 200 x; Bar = 100 μm. B. Representative images of tumors excised from Mcpt5-Cre− and Mcpt5-Cre+ mice 2 weeks after injection of 0.5 − 106 B16.F10 cells. Bar = 5 mm C. Tumor end weights 1 or 2 weeks after injection of 0.5 − 106 B16.F10 cells. Results are shown as mean ± SD; n = 4-7.CC BY · PubMed Central
Figure 2 Lungs from Mcpt-5 Cre+ mice retain expression of mucosal type mast cell markers and contain mast cells A. mRNA was prepared from lungs of Mcpt5-Cre− and Mcpt5-Cre+ mice and was analyzed by qPCR for content of transcripts for Mcpt1 (mucosal mast cell marker), Mcpt2 (mucosal mast cell marker) and Mcpt6 (marker of connective tissue type mast cells). Results are shown as mean values ± SD, n = 4, * p <0.05, ***p < 0.001. B. Representative images of lung sections from Mcpt5-Cre− and Mcpt5-Cre+ mice stained with toluidine blue. Note the presence of toluidine blue-positive mast cells in lungs of both Mcpt5-Cre− and Mcpt5-Cre+ mice. Original magnification = 400 x; Bar = 10 μm.
Figure 2 Lungs from Mcpt-5 Cre+ mice retain expression of mucosal type mast cell markers and contain mast cells A. mRNA was prepared from lungs of Mcpt5-Cre− and Mcpt5-Cre+ mice and was analyzed by qPCR for content of transcripts for Mcpt1 (mucosal mast cell marker), Mcpt2 (mucosal mast cell marker) and Mcpt6 (marker of connective tissue type mast cells). Results are shown as mean values ± SD, n = 4, * p <0.05, ***p < 0.001. B. Representative images of lung sections from Mcpt5-Cre− and Mcpt5-Cre+ mice stained with toluidine blue. Note the presence of toluidine blue-positive mast cells in lungs of both Mcpt5-Cre− and Mcpt5-Cre+ mice. Original magnification = 400 x; Bar = 10 μm.CC BY · PubMed Central
Figure 3 Reduced melanoma colonization in lungs from Mcpt5-Cre+ vs. Mcpt5-Cre− mice Melanoma cells (B16.F10) were intravenously injected into Mcpt5-Cre− or Mcpt5-Cre+ mice. After 11 days, lungs were excised. A. Representative images of lungs from Mcpt5-Cre− and Mcpt5-Cre+ mice. Note the abundance of melanoma nodules in lungs from Mcpt5-Cre− mice and the lower content of nodules in lungs from Mcpt5-Cre+ mice. B. Quantification of melanoma nodules; n = 8-9, ** p < 0.01. C. Representative image of lungs from Mcpt5-Cre− animals after melanoma administration. Melanoma areas are marked by arrowheads. D. Representative image of lungs from Mcpt5-Cre+ animals after melanoma administration. Melanoma accumulations are marked by arrowheads. E. Image showing the presence of mast cells (arrows; toluidine blue staining) in areas at a distance from the melanoma tumors (arrowhead). (C-E): Original magnification = 100 x; Bar = 100 μm. F. Representative image showing that mast cells occasionally are found in the immediate vicinity to melanoma tumors. G. Representative image showing a higher magnification of a lung tissue section from an Mcpt5-Cre− animal. Note the presence of highly granulated, toluidine blue-positive mast cells (arrows). H. Representative image showing a higher magnification of a lung tissue section from an Mcpt5-Cre+ animal. Note the presence of toluidine blue-positive mast cells (arrow). (F-H): Original magnification = 400 x; Bar = 50 μm. I. mRNA was extracted from lungs of melanoma-treated Mcpt5-Cre− or Mcpt5-Cre+ mice (+B16). As a control, mRNA was extracted from naïve animals (PBS). mRNA was analyzed by qPCR for the levels of transcripts for Dct2 and Gp100; mean values ± SEM; n = 4-6; * p < 0.05, ** p < 0.01. J. Correlation between the expression of Gp100 and Dct2. Regression analysis: R-sq (adj) = 98.7; p < 0.001; n = 10. K. Correlation between expression of Gp100 and number of melanoma nodules in lungs. Regression analysis: R-sq (adj) = 82.9; p < 0.001; n =10.
Figure 3 Reduced melanoma colonization in lungs from Mcpt5-Cre+ vs. Mcpt5-Cre− mice Melanoma cells (B16.F10) were intravenously injected into Mcpt5-Cre− or Mcpt5-Cre+ mice. After 11 days, lungs were excised. A. Representative images of lungs from Mcpt5-Cre− and Mcpt5-Cre+ mice. Note the abundance of melanoma nodules in lungs from Mcpt5-Cre− mice and the lower content of nodules in lungs from Mcpt5-Cre+ mice. B. Quantification of melanoma nodules; n = 8-9, ** p < 0.01. C. Representative image of lungs from Mcpt5-Cre− animals after melanoma administration. Melanoma areas are marked by arrowheads. D. Representative image of lungs from Mcpt5-Cre+ animals after melanoma administration. Melanoma accumulations are marked by arrowheads. E. Image showing the presence of mast cells (arrows; toluidine blue staining) in areas at a distance from the melanoma tumors (arrowhead). (C-E): Original magnification = 100 x; Bar = 100 μm. F. Representative image showing that mast cells occasionally are found in the immediate vicinity to melanoma tumors. G. Representative image showing a higher magnification of a lung tissue section from an Mcpt5-Cre− animal. Note the presence of highly granulated, toluidine blue-positive mast cells (arrows). H. Representative image showing a higher magnification of a lung tissue section from an Mcpt5-Cre+ animal. Note the presence of toluidine blue-positive mast cells (arrow). (F-H): Original magnification = 400 x; Bar = 50 μm. I. mRNA was extracted from lungs of melanoma-treated Mcpt5-Cre− or Mcpt5-Cre+ mice (+B16). As a control, mRNA was extracted from naïve animals (PBS). mRNA was analyzed by qPCR for the levels of transcripts for Dct2 and Gp100; mean values ± SEM; n = 4-6; * p < 0.05, ** p < 0.01. J. Correlation between the expression of Gp100 and Dct2. Regression analysis: R-sq (adj) = 98.7; p < 0.001; n = 10. K. Correlation between expression of Gp100 and number of melanoma nodules in lungs. Regression analysis: R-sq (adj) = 82.9; p < 0.001; n =10.CC BY · PubMed Central
Figure 4 Higher inflammation in bronchoalveolar lavage fluid of melanoma-treated Mcpt5-Cre+ vs. Mcpt5-Cre− animals Melanoma cells (B16.F10) were intravenously injected into Mcpt5-Cre− or Mcpt5-Cre+ littermate mice. After 11 days, bronchoalveolar lavage (BAL) was performed. As controls, BAL fluids from naïve Mcpt5-Cre− and Mcpt5-Cre+ mice were assessed. A-B. Total cell counts (A) and differential counting (B) of BAL fluid from either naïve or melanoma-treated Mcpt5-Cre− or Mcpt5-Cre+ mice. Results are given as mean ± SD; n = 4; * p < 0.05, ** p < 0.01. C. Representative images of May-Grünwald/Giemsa-stained cytospin slides prepared from BAL fluid of melanoma-treated Mcpt5-Cre− or Mcpt5-Cre+ mice. Note the presence of macrophages with ingested melanin (insets). Original magnification = 400 x; Bar = 10 μm.
Figure 4 Higher inflammation in bronchoalveolar lavage fluid of melanoma-treated Mcpt5-Cre+ vs. Mcpt5-Cre− animals Melanoma cells (B16.F10) were intravenously injected into Mcpt5-Cre− or Mcpt5-Cre+ littermate mice. After 11 days, bronchoalveolar lavage (BAL) was performed. As controls, BAL fluids from naïve Mcpt5-Cre− and Mcpt5-Cre+ mice were assessed. A-B. Total cell counts (A) and differential counting (B) of BAL fluid from either naïve or melanoma-treated Mcpt5-Cre− or Mcpt5-Cre+ mice. Results are given as mean ± SD; n = 4; * p < 0.05, ** p < 0.01. C. Representative images of May-Grünwald/Giemsa-stained cytospin slides prepared from BAL fluid of melanoma-treated Mcpt5-Cre− or Mcpt5-Cre+ mice. Note the presence of macrophages with ingested melanin (insets). Original magnification = 400 x; Bar = 10 μm.CC BY · PubMed Central
Figure 5 Altered expression of genes related to epithelial-mesenchymal transition in melanoma-treated Mcpt5-Cre− vs. Mcpt5-Cre+ mice Melanoma cells (B16.F10) were intravenously injected into Mcpt5-Cre− or Mcpt5-Cre+ mice. After 11 days, lungs were excised, followed by preparation of mRNA and qPCR analysis of levels of transcripts for Snail, Slug, Twist, Zeb1, Zeb2, E-cadherin, Vegf and Il2. Results are presented as mean ± SEM; n = 5-6, * p < 0.05.
Figure 5 Altered expression of genes related to epithelial-mesenchymal transition in melanoma-treated Mcpt5-Cre− vs. Mcpt5-Cre+ mice Melanoma cells (B16.F10) were intravenously injected into Mcpt5-Cre− or Mcpt5-Cre+ mice. After 11 days, lungs were excised, followed by preparation of mRNA and qPCR analysis of levels of transcripts for Snail, Slug, Twist, Zeb1, Zeb2, E-cadherin, Vegf and Il2. Results are presented as mean ± SEM; n = 5-6, * p < 0.05.CC BY · PubMed Central

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Metadata

Authors
Öhrvik, H., Grujic, M., Waern, I., Gustafson, A.-M., Ernst, N., Roers, A., Hartmann, K., & Pejler, G.
Affiliations
  • Department of Medical Biochemistry and Microbiology, Uppsala University, Uppsala, Sweden
  • Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, Uppsala, Sweden
  • Department of Dermatology, University of Luebeck, Luebeck, Germany
  • Institute for Immunology, University of Technology Dresden, Dresden, Germany
Venue
Oncotarget
Publisher
Impact Journals, LLC
Published
2016-09-02
Volume
7
Issue
42
Pages
68990-69001
ISSN
1949-2553
DOI
10.18632/oncotarget.11837
PMC
PMC5356606
Cited by
28
References
1
Language
en

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CoChat. (2026). Citation check: Mast cells promote melanoma colonization of lungs. https://app.cochat.ai/papers/10.18632/oncotarget.11837
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Öhrvik, H., Grujic, M., Waern, I., Gustafson, A.-M., Ernst, N., Roers, A., Hartmann, K., & Pejler, G. (2016). Mast cells promote melanoma colonization of lungs. Oncotarget. https://doi.org/10.18632/oncotarget.11837