EMMPRIN regulates β1 integrin-mediated adhesion through Kindlin-3 in human melanoma cells

EMMPRIN is known to promote tumor invasion through extracellular matrix (ECM) degradation. Here we report that EMMPRIN can regulate melanoma cell adhesion to the ECM through an interaction with β1 integrin involving kindlin‐3. In this study, EMMPRIN knockdown in the human melanoma cell line M10 usin...

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Published inExperimental dermatology Vol. 24; no. 6; pp. 443 - 448
Main Authors Delyon, Julie, Khayati, Farah, Djaafri, Ibtissem, Podgorniak, Marie-Pierre, Sadoux, Aurélie, Setterblad, Niclas, Boutalbi, Zineb, Maouche, Kamel, Maskos, Uwe, Menashi, Suzanne, Lebbé, Céleste, Mourah, Samia
Format Journal Article
LanguageEnglish
Published Denmark Blackwell Publishing Ltd 01.06.2015
Wiley
Subjects
Animals
Antigens, CD147
Antigens, CD29
Basigin - drug effects
Basigin - genetics
Basigin - physiology
Cell Adhesion
Cell Adhesion - physiology
Cell Line, Tumor
Cell Shape
Cell Shape - physiology
Cognitive science
EMMPRIN
Extracellular Matrix
Extracellular Matrix - physiology
Female
Gene Expression Regulation, Neoplastic
Gene Expression Regulation, Neoplastic - drug effects
Gene Knockdown Techniques
Heterografts
Humans
In Vitro Techniques
Integrin beta1 - physiology
Kindlin-3
Life Sciences
Melanoma
Melanoma - pathology
Melanoma - physiopathology
Membrane Proteins
Membrane Proteins - physiology
Mice
Mice, Nude
Neoplasm Proteins
Neoplasm Proteins - physiology
Neuroscience
RNA, Small Interfering
RNA, Small Interfering - pharmacology
Signal Transduction
Signal Transduction - physiology
Skin Neoplasms
Skin Neoplasms - pathology
Skin Neoplasms - physiopathology
β1 integrin
EMMPRIN
melanoma
β1 integrin
cell adhesion
Kindlin-3
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Summary:EMMPRIN is known to promote tumor invasion through extracellular matrix (ECM) degradation. Here we report that EMMPRIN can regulate melanoma cell adhesion to the ECM through an interaction with β1 integrin involving kindlin‐3. In this study, EMMPRIN knockdown in the human melanoma cell line M10 using siRNA decreased cell invasion and significantly increased cell adhesion and spreading. A morphological change from a round to a spread shape was observed associated with enhanced phalloidin‐labelled actin staining. In situ proximity ligation assay and co‐immunoprecipitation revealed that EMMPRIN silencing increased the interaction of β1 integrin with kindlin‐3, a focal adhesion protein. This was associated with an increase in β1 integrin activation and a decrease in the phosphorylation of the downstream integrin kinase FAK. Moreover, the expression at both the transcript and protein level of kindlin‐3 and of β1 integrin was inversely regulated by EMMPRIN. EMMPRIN did not regulate either talin expression or its interaction with β1 integrin. These results are consistent with our in vivo demonstration that EMMPRIN inhibition increased β1 integrin activation and its interaction with kindlin‐3. To conclude, these findings reveal a new role of EMMPRIN in tumor cell migration through ß1 integrin/kindlin‐3‐mediated adhesion pathway.
Bibliography:Ligue Nationale contre le Cancer
ark:/67375/WNG-BCZZPFR8-C
Institut National de la Santé et de la Recherche Médicale (INSERM)
istex:BDDE6F6E35C12BF7D5CF8201BB6292790FBA07B5
Assistance Publique-Hôpitaux de Paris (APHP) as a Contrat Année-Recherche
ArticleID:EXD12693
Société Française de Dermatologie (SFD)
Figure S1. EMMPRIN knockdown inhibits the tumor growth in melanoma xenografts. (a) Western blot analyses of EMMPRIN in control miRNA and EMMPRIN miRNA BLM melanoma cells, prior to xenografting in mice. Equal loading of proteins was assessed by probing for β-actin. (b) Stable EMMPRIN silencing by miRNA in BLM cells decreased tumor growth. Representative photos of tumors 5 weeks after injection (5 mice per group). Results represent mean of tumor volume ± SD. Figure S2. EMMPRIN down-regulation in EMMPRIN siRNA-transfected melanoma cells. Left, relative expression of EMMPRIN mRNA to the reference gene β2-microglobulin (β2m) assessed by qRT-PCR. Results represent mean ± SD. Right, western blot analyses of EMMPRIN in control-siRNA and EMMPRIN-siRNA transfected M10 cells. Equal loading of proteins was assessed by probing for β-actin. Figure S3. EMMPRIN down-regulation inhibits melanoma cell invasion. SiRNA-mediated silencing of EMMPRIN expression. Left, invaded transfected melanoma cells M10 through Matrigel after 24 h; right, invaded cells counted in 6 microscopic fields per sample. Results represent the average of duplicate samples from three independent experiments; Bars, SD. *P < 0.05. Figure S4. EMMPRIN regulates β1 integrin and kindlin-3 expression. B1 integrin and kindlin-3 expression analysed with western blot in control- and EMMPRIN-siRNA transfected cells. Equal loading of proteins was assessed by probing for β-actin. Figure S5.β1 integrin/talin interaction is not regulated by EMMPRIN. In control- and EMMPRIN-siRNA transfected melanoma cells, immunoprecipitation of β1 integrin followed by immunoblotting with the talin antibody. Equal loading of proteins was assessed by probing for β-actin.
ISSN:0906-6705
1600-0625
DOI:10.1111/exd.12693