Morphine Modulates Interleukin-4- or Breast Cancer Cell-induced Pro-metastatic Activation of Macrophages
Identifieur interne : 000803 ( Pmc/Curation ); précédent : 000802; suivant : 000804Morphine Modulates Interleukin-4- or Breast Cancer Cell-induced Pro-metastatic Activation of Macrophages
Auteurs : Samira Khabbazi [Australie] ; Yannick Goumon [France] ; Marie-Odile Parat [Australie]Source :
- Scientific Reports [ 2045-2322 ] ; 2015.
Abstract
Interactions between cancer cells and stromal cells in the tumour microenvironment play a key role in the control of invasiveness, metastasis and angiogenesis. Macrophages display a range of activation states in specific pathological contexts and alternatively activated (M2) macrophages can promote tumour aggressiveness. Opioids are able to modulate tumour growth and metastasis. We tested whether morphine modulates the activation of macrophages induced by (i) interleukin-4 (IL-4), the prototypical M2 polarization-inducing cytokine, or (ii) coculture with breast cancer cells. We showed that IL-4 causes increased MMP-9 production and expression of the alternative activation markers arginase-1 and MRC-1. Morphine prevented IL-4-induced increase in MMP-9 in a naloxone- and methylnaltrexone-reversible fashion. Morphine also prevented IL-4-elicited alternative activation of RAW264.7 macrophages. Expression of MMP-9 and arginase-1 were increased when RAW264.7 were subjected to paracrine activation by 4T1 cells, and this effect was prevented by morphine via an opioid receptor-mediated mechanism. Morphine further decreased 4T1 breast cancer cell invasion elicited by co-culture with RAW264.7. Reduction of MMP-9 expression and alternative activation of macrophages by morphine was confirmed using mouse bone marrow-derived macrophages. Taken together, our results indicate that morphine may modulate tumour aggressiveness by regulating macrophage protease production and M2 polarization within the tumour microenvironment.
Url:
DOI: 10.1038/srep11389
PubMed: 26078009
PubMed Central: 4468425
Links toward previous steps (curation, corpus...)
- to stream Pmc, to step Corpus: Pour aller vers cette notice dans l'étape Curation :000804
Links to Exploration step
PMC:4468425Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Morphine Modulates Interleukin-4- or Breast Cancer Cell-induced Pro-metastatic Activation of Macrophages</title><author><name sortKey="Khabbazi, Samira" sort="Khabbazi, Samira" uniqKey="Khabbazi S" first="Samira" last="Khabbazi">Samira Khabbazi</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>University of Queensland School of Pharmacy, PACE</institution>, 20 Cornwall Street, Woollloongabba QLD 4102,<country>Australia</country></nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Goumon, Yannick" sort="Goumon, Yannick" uniqKey="Goumon Y" first="Yannick" last="Goumon">Yannick Goumon</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>CNRS UPR3212, Institut des Neurosciences Cellulaires et Intégratives, Centre National de la Recherche Scientifique and University of Strasbourg</institution>, 5 rue Blaise Pascal, 67084 Strasbourg,<country>France</country></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Parat, Marie Odile" sort="Parat, Marie Odile" uniqKey="Parat M" first="Marie-Odile" last="Parat">Marie-Odile Parat</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>University of Queensland School of Pharmacy, PACE</institution>, 20 Cornwall Street, Woollloongabba QLD 4102,<country>Australia</country></nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">26078009</idno><idno type="pmc">4468425</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4468425</idno><idno type="RBID">PMC:4468425</idno><idno type="doi">10.1038/srep11389</idno><date when="2015">2015</date><idno type="wicri:Area/Pmc/Corpus">000804</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000804</idno><idno type="wicri:Area/Pmc/Curation">000803</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000803</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Morphine Modulates Interleukin-4- or Breast Cancer Cell-induced Pro-metastatic Activation of Macrophages</title><author><name sortKey="Khabbazi, Samira" sort="Khabbazi, Samira" uniqKey="Khabbazi S" first="Samira" last="Khabbazi">Samira Khabbazi</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>University of Queensland School of Pharmacy, PACE</institution>, 20 Cornwall Street, Woollloongabba QLD 4102,<country>Australia</country></nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Goumon, Yannick" sort="Goumon, Yannick" uniqKey="Goumon Y" first="Yannick" last="Goumon">Yannick Goumon</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>CNRS UPR3212, Institut des Neurosciences Cellulaires et Intégratives, Centre National de la Recherche Scientifique and University of Strasbourg</institution>, 5 rue Blaise Pascal, 67084 Strasbourg,<country>France</country></nlm:aff><country xml:lang="fr">France</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Parat, Marie Odile" sort="Parat, Marie Odile" uniqKey="Parat M" first="Marie-Odile" last="Parat">Marie-Odile Parat</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>University of Queensland School of Pharmacy, PACE</institution>, 20 Cornwall Street, Woollloongabba QLD 4102,<country>Australia</country></nlm:aff><country xml:lang="fr">Australie</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></analytic><series><title level="j">Scientific Reports</title><idno type="eISSN">2045-2322</idno><imprint><date when="2015">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Interactions between cancer cells and stromal cells in the tumour microenvironment play a key role in the control of invasiveness, metastasis and angiogenesis. Macrophages display a range of activation states in specific pathological contexts and alternatively activated (M2) macrophages can promote tumour aggressiveness. Opioids are able to modulate tumour growth and metastasis. We tested whether morphine modulates the activation of macrophages induced by (i) interleukin-4 (IL-4), the prototypical M2 polarization-inducing cytokine, or (ii) coculture with breast cancer cells. We showed that IL-4 causes increased MMP-9 production and expression of the alternative activation markers arginase-1 and MRC-1. Morphine prevented IL-4-induced increase in MMP-9 in a naloxone- and methylnaltrexone-reversible fashion. Morphine also prevented IL-4-elicited alternative activation of RAW264.7 macrophages. Expression of MMP-9 and arginase-1 were increased when RAW264.7 were subjected to paracrine activation by 4T1 cells, and this effect was prevented by morphine via an opioid receptor-mediated mechanism. Morphine further decreased 4T1 breast cancer cell invasion elicited by co-culture with RAW264.7. Reduction of MMP-9 expression and alternative activation of macrophages by morphine was confirmed using mouse bone marrow-derived macrophages. Taken together, our results indicate that morphine may modulate tumour aggressiveness by regulating macrophage protease production and M2 polarization within the tumour microenvironment.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Afsharimani, B" uniqKey="Afsharimani B">B. Afsharimani</name></author><author><name sortKey="Cabot, P" uniqKey="Cabot P">P. Cabot</name></author><author><name sortKey="Parat, M O" uniqKey="Parat M">M. O. Parat</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Afsharimani, B" uniqKey="Afsharimani B">B. Afsharimani</name></author><author><name sortKey="Cabot, P J" uniqKey="Cabot P">P. J. Cabot</name></author><author><name sortKey="Parat, M O" uniqKey="Parat M">M. O. Parat</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Shavit, Y" uniqKey="Shavit Y">Y. Shavit</name></author><author><name sortKey="Ben Eliyahu, S" uniqKey="Ben Eliyahu S">S. Ben-Eliyahu</name></author><author><name sortKey="Zeidel, A" uniqKey="Zeidel A">A. Zeidel</name></author><author><name sortKey="Beilin, B" uniqKey="Beilin B">B. Beilin</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Gupta, K" uniqKey="Gupta K">K. Gupta</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lennon, F E" uniqKey="Lennon F">F. E. Lennon</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Page, G G" uniqKey="Page G">G. G. Page</name></author><author><name sortKey="Blakely, W P" uniqKey="Blakely W">W. P. Blakely</name></author><author><name sortKey="Ben Eliyahu, S" uniqKey="Ben Eliyahu S">S. Ben-Eliyahu</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Koodie, L" uniqKey="Koodie L">L. Koodie</name></author><author><name sortKey="Ramakrishnan, S" uniqKey="Ramakrishnan S">S. Ramakrishnan</name></author><author><name sortKey="Roy, S" uniqKey="Roy S">S. Roy</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Afsharimani, B" uniqKey="Afsharimani B">B. Afsharimani</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Sica, A" uniqKey="Sica A">A. Sica</name></author><author><name sortKey="Mantovani, A" uniqKey="Mantovani A">A. Mantovani</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wang, H W" uniqKey="Wang H">H. W. Wang</name></author><author><name sortKey="Joyce, J A" uniqKey="Joyce J">J. A. Joyce</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Denardo, D G" uniqKey="Denardo D">D. G. DeNardo</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Franklin, R A" uniqKey="Franklin R">R. A. Franklin</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Vandooren, J" uniqKey="Vandooren J">J. Vandooren</name></author><author><name sortKey="Van Den Steen, P E" uniqKey="Van Den Steen P">P. E. Van den Steen</name></author><author><name sortKey="Opdenakker, G" uniqKey="Opdenakker G">G. Opdenakker</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Galdiero, M R" uniqKey="Galdiero M">M. R. Galdiero</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chizzolini, C" uniqKey="Chizzolini C">C. Chizzolini</name></author><author><name sortKey="Rezzonico, R" uniqKey="Rezzonico R">R. Rezzonico</name></author><author><name sortKey="De Luca, C" uniqKey="De Luca C">C. De Luca</name></author><author><name sortKey="Burger, D" uniqKey="Burger D">D. Burger</name></author><author><name sortKey="Dayer, J M" uniqKey="Dayer J">J. M. Dayer</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Zhang, Y" uniqKey="Zhang Y">Y. Zhang</name></author><author><name sortKey="Mccluskey, K" uniqKey="Mccluskey K">K. McCluskey</name></author><author><name sortKey="Fujii, K" uniqKey="Fujii K">K. Fujii</name></author><author><name sortKey="Wahl, L M" uniqKey="Wahl L">L. M. Wahl</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lacraz, S" uniqKey="Lacraz S">S. Lacraz</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Corcoran, M L" uniqKey="Corcoran M">M. L. Corcoran</name></author><author><name sortKey="Stetler Stevenson, W G" uniqKey="Stetler Stevenson W">W. G. Stetler-Stevenson</name></author><author><name sortKey="Brown, P D" uniqKey="Brown P">P. D. Brown</name></author><author><name sortKey="Wahl, L M" uniqKey="Wahl L">L. M. Wahl</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Webster, N L" uniqKey="Webster N">N. L. Webster</name></author><author><name sortKey="Crowe, S M" uniqKey="Crowe S">S. M. Crowe</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lolmede, K" uniqKey="Lolmede K">K. Lolmede</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Doornebal, C W" uniqKey="Doornebal C">C. W. Doornebal</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chen, W" uniqKey="Chen W">W. Chen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Godai, K" uniqKey="Godai K">K. Godai</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chang, C I" uniqKey="Chang C">C. I. Chang</name></author><author><name sortKey="Liao, J C" uniqKey="Liao J">J. C. Liao</name></author><author><name sortKey="Kuo, L" uniqKey="Kuo L">L. Kuo</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chang, C I L" uniqKey="Chang C">C. I. L. Chang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Crittenden, M R" uniqKey="Crittenden M">M. R. Crittenden</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Joyce, J A" uniqKey="Joyce J">J. A. Joyce</name></author><author><name sortKey="Pollard, J W" uniqKey="Pollard J">J. W. Pollard</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Gocheva, V" uniqKey="Gocheva V">V. Gocheva</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Gach, K" uniqKey="Gach K">K. Gach</name></author><author><name sortKey="Szemraj, J" uniqKey="Szemraj J">J. Szemraj</name></author><author><name sortKey="Wyrebska, A" uniqKey="Wyrebska A">A. Wyrebska</name></author><author><name sortKey="Janecka, A" uniqKey="Janecka A">A. Janecka</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Juneja, R" uniqKey="Juneja R">R. Juneja</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Schmittgen, T D" uniqKey="Schmittgen T">T. D. Schmittgen</name></author><author><name sortKey="Livak, K J" uniqKey="Livak K">K. J. Livak</name></author></analytic></biblStruct></listBibl></div1></back></TEI><pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Sci Rep</journal-id><journal-id journal-id-type="iso-abbrev">Sci Rep</journal-id><journal-title-group><journal-title>Scientific Reports</journal-title></journal-title-group><issn pub-type="epub">2045-2322</issn><publisher><publisher-name>Nature Publishing Group</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">26078009</article-id><article-id pub-id-type="pmc">4468425</article-id><article-id pub-id-type="pii">srep11389</article-id><article-id pub-id-type="doi">10.1038/srep11389</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>Morphine Modulates Interleukin-4- or Breast Cancer Cell-induced Pro-metastatic Activation of Macrophages</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Khabbazi</surname><given-names>Samira</given-names></name><xref ref-type="aff" rid="a1">1</xref></contrib><contrib contrib-type="author"><name><surname>Goumon</surname><given-names>Yannick</given-names></name><xref ref-type="aff" rid="a2">2</xref></contrib><contrib contrib-type="author"><name><surname>Parat</surname><given-names>Marie-Odile</given-names></name><xref ref-type="corresp" rid="c1">a</xref><xref ref-type="aff" rid="a1">1</xref></contrib><aff id="a1"><label>1</label><institution>University of Queensland School of Pharmacy, PACE</institution>, 20 Cornwall Street, Woollloongabba QLD 4102,<country>Australia</country></aff><aff id="a2"><label>2</label><institution>CNRS UPR3212, Institut des Neurosciences Cellulaires et Intégratives, Centre National de la Recherche Scientifique and University of Strasbourg</institution>, 5 rue Blaise Pascal, 67084 Strasbourg,<country>France</country></aff></contrib-group><author-notes><corresp id="c1"><label>a</label><email>m.parat@uq.edu.au</email></corresp></author-notes><pub-date pub-type="epub"><day>16</day><month>06</month><year>2015</year></pub-date><pub-date pub-type="collection"><year>2015</year></pub-date><volume>5</volume><elocation-id>11389</elocation-id><history><date date-type="received"><day>03</day><month>02</month><year>2015</year></date><date date-type="accepted"><day>15</day><month>05</month><year>2015</year></date></history><permissions><copyright-statement>Copyright © 2015, Macmillan Publishers Limited</copyright-statement><copyright-year>2015</copyright-year><copyright-holder>Macmillan Publishers Limited</copyright-holder><license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by/4.0/"><pmc-comment>author-paid</pmc-comment>
<license-p>This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">http://creativecommons.org/licenses/by/4.0/</ext-link></license-p></license></permissions><abstract><p>Interactions between cancer cells and stromal cells in the tumour microenvironment play a key role in the control of invasiveness, metastasis and angiogenesis. Macrophages display a range of activation states in specific pathological contexts and alternatively activated (M2) macrophages can promote tumour aggressiveness. Opioids are able to modulate tumour growth and metastasis. We tested whether morphine modulates the activation of macrophages induced by (i) interleukin-4 (IL-4), the prototypical M2 polarization-inducing cytokine, or (ii) coculture with breast cancer cells. We showed that IL-4 causes increased MMP-9 production and expression of the alternative activation markers arginase-1 and MRC-1. Morphine prevented IL-4-induced increase in MMP-9 in a naloxone- and methylnaltrexone-reversible fashion. Morphine also prevented IL-4-elicited alternative activation of RAW264.7 macrophages. Expression of MMP-9 and arginase-1 were increased when RAW264.7 were subjected to paracrine activation by 4T1 cells, and this effect was prevented by morphine via an opioid receptor-mediated mechanism. Morphine further decreased 4T1 breast cancer cell invasion elicited by co-culture with RAW264.7. Reduction of MMP-9 expression and alternative activation of macrophages by morphine was confirmed using mouse bone marrow-derived macrophages. Taken together, our results indicate that morphine may modulate tumour aggressiveness by regulating macrophage protease production and M2 polarization within the tumour microenvironment.</p></abstract></article-meta></front><floats-group><fig id="f1"><label>Figure 1</label><caption><title>Alternative activation of RAW264.7 macrophages by IL-4.</title><p><bold>A</bold>) Production of gelatinases MMP-9 and MMP-2 in the conditioned medium of RAW264.7 cells treated with indicated concentrations of IL-4 for 48 h was assessed by gelatine zymography. <bold>B</bold>) RAW264.7 cells were treated with 5 ng/ml IL-4 for 48 h and gelatinase production determined by zymography followed by densitometric quantitation of MMP-9 and MMP-2. Mean ± SEM is shown, n = 3 independent experiments. ***p < 0.001; ns, no statistically significant difference, Student’s <italic>t</italic>-test <bold>C</bold>) Real-time RT-PCR determination of mRNA levels of MMP-9 as well as alternative activation markers MRC-1 and arginase-1 (Arg-1) in RAW264.7 cells treated with 5 ng/ml IL-4 for 48h. Results are expressed relative to untreated control. Mean ± SEM is shown, n = 7–10 independent experiments. *p < 0.05, **p < 0.01, Student’s <italic>t</italic>-test.</p></caption><graphic xlink:href="srep11389-f1"></graphic></fig><fig id="f2"><label>Figure 2</label><caption><title>Morphine inhibits IL-4 induced increase of MMP-9 production by RAW264.7 macrophages.</title><p><bold>A</bold>) RAW264.7 macrophages were incubated with IL-4 (5 ng/ml) in the presence or absence of morphine (20 μM). Control cells were incubated with buffer alone. Conditioned media were tested for gelatinase production by zymography. <bold>B</bold>) Zymography gels were subjected to densitometric quantitation of MMP-9. Mean ± SEM is shown, n = 3 separate experiments. ****p < 0.0001, Student’s <italic>t</italic>-test. ; <bold>C</bold>) Real-time RT-PCR determination of mRNA levels of MMP-9 and alternative activation marker arginase-1 (Arg-1) or MRC-1 in RAW264.7 cells treated with 5 ng/ml IL-4 and either 0, 10 or 20 μM morphine for 48 h. Results are expressed relative to IL-4 only treated cells. Mean ± SEM is shown, n = 3–6 independent experiments. *p < 0.05; **p < 0.01; ns, no statistical significance, One way ANOVA with Dunnet’s multiple comparison test. <bold>D</bold>) Real-time RT-PCR determination of mRNA levels of MMP-9 or alternative activation marker arginase-1 (Arg-1) in RAW264.7 cells treated with 0, 10 or 20 μM morphine for 48h. Results are expressed relative to untreated control. Mean ± SEM is shown, n = 4 independent experiments. ns, no statistical significance; *p < 0.05, One way ANOVA with Dunnet’s multiple comparison test.</p></caption><graphic xlink:href="srep11389-f2"></graphic></fig><fig id="f3"><label>Figure 3</label><caption><title>The effect of morphine is opioid receptor-mediated.</title><p>RAW264.7 macrophages were incubated for 48`h with IL-4 (5 ng/ml) in the presence or absence of morphine (20 μM) and naloxone (20 μM) or methylnaltrexone (MNTX) (20 μM) as indicated. <bold>A</bold>) Conditioned media were collected and analysed by gelatine zymography. <bold>B</bold>) Zymography gels were used for densitometric quantitation. Mean ± SEM is shown, n = 5 independent experiments. *p < 0.05; ***p < 0.001; ****p < 0.0001, Student’s <italic>t</italic> test. <bold>C</bold>) Real-time RT-PCR determination of mRNA levels of MMP-9 in RAW264.7 cells treated as indicated. Results are expressed relative to IL-4 only treated cells. Mean ± SEM is shown, n = 5 independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001, Student’s <italic>t</italic> test.</p></caption><graphic xlink:href="srep11389-f3"></graphic></fig><fig id="f4"><label>Figure 4</label><caption><title>MMP-9 production is increased in Transwell® co-cultures of macrophages and 4T1 cells.</title><p>RAW264.7 macrophages (in the lower chamber) and 4T1 cells (in the upper chamber) were cultured individually or together for 48 h in 12-well Transwell®. <bold>A</bold>) Conditioned media were collected and production of MMP-9 was determined by gelatine zymography. <bold>B</bold>) Densitometric quantitation of MMP-9 activity in the conditioned media. Mean ± SEM is shown. n = 3 independent experiments ***p < 0.001 compared to 4T1 or RAW264.7 alone, Student’s <italic>t</italic> test. <bold>C-D</bold>) Real-time PCR determination of MMP-9 mRNA levels in RAW264.7 cells (<bold>C</bold>) and 4T1 cells (<bold>D</bold>) alone or co-cultured for 48h. Results are expressed relative to each cell type grown individually. Mean ± SEM is shown, n = 3 independent experiments. *p < 0.05, Student’s <italic>t</italic> test. <bold>E</bold>) Real-time PCR determination of arginase-1 mRNA levels of RAW264.7 cells co-cultured with 4T1 cells for 48 h. Results are expressed relative to RAW264.7 grown individually. Mean ± SEM is shown, n = 3 independent experiments. **p < 0.01, Student’s <italic>t</italic> test.</p></caption><graphic xlink:href="srep11389-f4"></graphic></fig><fig id="f5"><label>Figure 5</label><caption><title>Morphine inhibits coculture-induced increase in MMP-9 production by RAW264.7 macrophages.</title><p>RAW264.7 macrophages (in the lower chamber) and 4T1 cells (in the upper chamber) were cultured individually or together for 48 h in 12-well Transwell® in the presence (10 or 20 μM as indicated) or absence of morphine. <bold>A</bold>) Production of MMP-9 in the conditioned media was determined by gelatine zymography. <bold>B</bold>) Densitometric quantitation of MMP-9 activity in the conditioned media. Results are expressed as a % of MMP9 in 4T1 alone. Mean ± SEM is shown. n = 3 independent experiments *p < 0.05; ***p < 0.001, Two way ANOVA with Tukey’s multiple comparisons test. <bold>C</bold>) Real-time PCR determination of MMP-9 mRNA levels in RAW264.7 cells co-cultured for 48 h with 4T1 in the presence (10 or 20 μM as indicated) or absence of morphine. Results are expressed relative to RAW264.7 cells co-cultured with 4T1 in the absence of morphine. Mean ± SEM is shown, n = 3 independent experiments. *p < 0.05, One way ANOVA with Dunnet’s multiple comparisons test. <bold>D</bold>) Real-time PCR determination of arginase-1 (Arg-1) mRNA levels in RAW264.7 cells co-cultured for 48 h with 4T1 in the presence (10 or 20 μM as indicated) or absence of morphine. Results are expressed relative to RAW264.7 cells co-cultured with 4T1 in the absence of morphine. Mean ± SEM is shown, n = 3 independent experiments. ***p < 0.001. ****p < 0.0001, One way ANOVA with Dunnet’s multiple comparisons test. <bold>E–F</bold>, RAW264.7 macrophages (in the lower chamber) and 4T1 cells (in the upper chamber) were cultured together for 48 h in 12-well Transwell® in the presence of morphine (20 μM) and naloxone (20 μM) or methylnaltrexone (MNTX) (20 μM) as indicated. <bold>E</bold>) Conditioned media were collected and analysed by gelatine zymography. <bold>F</bold>) Zymography gels were used for densitometric quantitation. Mean ± SEM is shown, n = 3 independent experiments. *p < 0.05; ***p < 0.001, Student’s <italic>t</italic> test.</p></caption><graphic xlink:href="srep11389-f5"></graphic></fig><fig id="f6"><label>Figure 6</label><caption><title>Morphine prevents macrophage-induced breast cancer cell invasion.</title><p>Invasion of matrigel-coated inserts by 4T1 cells over 48 h was allowed to proceed in the absence or presence of RAW264.7 cells seeded in the bottom well of the chamber, and of 20 μM morphine as indicated. <bold>A</bold>) Representative images of invaded cells after fixation and staining. <bold>B</bold>) Quantification of invaded cells. Mean ± SEM is shown, n = 3 independent experiments. **p < 0.01; ***p < 0.001, Student’s <italic>t</italic> test.</p></caption><graphic xlink:href="srep11389-f6"></graphic></fig><fig id="f7"><label>Figure 7</label><caption><title>Morphine inhibits alternative activation of bone marrow-derived macrophages (BMM).</title><p><bold>A</bold>) BMM were treated with 5 ng/ml IL-4 for 24 h and mRNA expression of MMP-9 and the M2 polarization markers Arg-1 and MRC-1 was assessed by real-time RT-PCR. Results are expressed relative to untreated control. Mean ± SEM is shown, n = 4 independent experiments. **p < 0.01, ***p < 0.001, Student’s <italic>t</italic>-test. <bold>B</bold>) BMM were incubated with IL-4 (5 ng/ml) in the presence or absence of morphine at the indicated concentrations (up to 20 μM) for 24 h and mRNA expression of Arg-1 MMP-9 or MRC-1 was determined by real time RT-PCR. Results are expressed relative to IL-4 only treated cells. Mean ± SEM is shown, n = 3–4 independent experiments. *p < 0.05; **p < 0.01 ns, no statistical significance, One way ANOVA with Dunnet’s multiple comparison test. <bold>C)</bold> BMM and 4T1 cells were cultured individually or together for 24 h in 12-well Transwell® and mRNA levels of Arg-1, MMP-9 and MRC-1 determined by real time RT-PCR in BMM alone or co-cultured with 4T1 cells. Results are expressed relative to BMM grown individually. Mean ± SEM is shown, n = 3–4 independent experiments. *p < 0.05, Student’s <italic>t</italic> test. <bold>D</bold>) BMM were co-cultured with 4T1 in the presence or absence of morphine at the indicated concentrations (up to 20 μM) for 24 h and mRNA expression of Arg-1 MMP-9 or MRC-1 determined by real time RT-PCR. Results are expressed relative to BMM co-cultured with 4T1 in the absence of morphine. Mean ± SEM is shown, n = 3–4 independent experiments. *p < 0.05; **p < 0.01, ***p < 0.001, ns, no statistical significance, One way ANOVA with Dunnet’s multiple comparison test.</p></caption><graphic xlink:href="srep11389-f7"></graphic></fig></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Wicri/Asie/explor/AustralieFrV1/Data/Pmc/Curation
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000803 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Pmc/Curation/biblio.hfd -nk 000803 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Wicri/Asie
|area= AustralieFrV1
|flux= Pmc
|étape= Curation
|type= RBID
|clé= PMC:4468425
|texte= Morphine Modulates Interleukin-4- or Breast Cancer Cell-induced Pro-metastatic Activation of Macrophages
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Pmc/Curation/RBID.i -Sk "pubmed:26078009" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Pmc/Curation/biblio.hfd \
| NlmPubMed2Wicri -a AustralieFrV1
| This area was generated with Dilib version V0.6.33. |  |