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Tumor-derived microvesicles mediate human breast cancer invasion through differentially glycosylated EMMPRIN.

Menck K, Scharf C, Bleckmann A, Dyck L, Rost U, Wenzel D, Dhople VM, Siam L, Pukrop T, Binder C, Klemm F - J Mol Cell Biol (2014)

Bottom Line: Uptake of T-MV is essential for the proinvasive effect.Interestingly, the effect of EMMPRIN-bearing MV is not mediated by matrix metalloproteinases but by activation of the p38/MAPK signaling pathway in the tumor cells.In conclusion, T-MV stimulate cancer cell invasion via a direct feedback mechanism dependent on highly glycosylated EMMPRIN.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology/Oncology, University Medical Center Göttingen, 37099 Göttingen, Germany.

No MeSH data available.


Related in: MedlinePlus

High glycosylation is essential for the proinvasive function of EMMPRIN. (A) The EMMPRIN glycosylation status on tumor and benign cells/MV was analyzed by western blotting (HG-EMP, highly glycosylated EMMPRIN; IG-EMP, intermediately glycosylated EMMPRIN; LG-EMP, lowly glycosylated EMMPRIN). (B) T-MVM were deglycosylated by PNGaseF (visible as an unspecific band in lanes 1 and 3) under denaturing or nondenaturing conditions and the EMMPRIN glycosylation status was assessed by western blotting. (C) Microinvasion assay of MCF-7 cells stimulated with deglycosylated T-MV (10 µg/ml, non-denat. conditions) (mean ± SD, n = 3, *P < 0.001).
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MJU047F4: High glycosylation is essential for the proinvasive function of EMMPRIN. (A) The EMMPRIN glycosylation status on tumor and benign cells/MV was analyzed by western blotting (HG-EMP, highly glycosylated EMMPRIN; IG-EMP, intermediately glycosylated EMMPRIN; LG-EMP, lowly glycosylated EMMPRIN). (B) T-MVM were deglycosylated by PNGaseF (visible as an unspecific band in lanes 1 and 3) under denaturing or nondenaturing conditions and the EMMPRIN glycosylation status was assessed by western blotting. (C) Microinvasion assay of MCF-7 cells stimulated with deglycosylated T-MV (10 µg/ml, non-denat. conditions) (mean ± SD, n = 3, *P < 0.001).

Mentions: In order to test whether differential EMMPRIN expression could explain the proinvasive effect of T-MV in contrast to benign MV, we analyzed EMMPRIN expression in various cells and their MV. EMMPRIN was strongly enriched in T-MV compared with their parental cells, but only weakly expressed on platelet-derived MV (P-MV) (Figure 4A) that were previously identified as being not proinvasive (Menck et al., 2013). Surprisingly, the strongest EMMPRIN enrichment was demonstrated on MV from the benign hTERT-HME1. Analysis of the protein size revealed that all investigated MV contained a large form of EMMPRIN ranging from 45 to 65 kDa, indicating glycosylation at multiple sites. However, within this fraction, two different glycoforms can be distinguished: (i) a very highly glycosylated protein (HG-EMMPRIN, ∼50–65 kDa) characterizing all proinvasive MV, and (ii) a variant with intermediate glycosylation (IG-EMMPRIN, ∼45 kDa) in all non-proinvasive MV. The low glycosylation variant (LG-EMMPRIN, 32 kDa) was only very weakly detectable.Figure 4


Tumor-derived microvesicles mediate human breast cancer invasion through differentially glycosylated EMMPRIN.

Menck K, Scharf C, Bleckmann A, Dyck L, Rost U, Wenzel D, Dhople VM, Siam L, Pukrop T, Binder C, Klemm F - J Mol Cell Biol (2014)

High glycosylation is essential for the proinvasive function of EMMPRIN. (A) The EMMPRIN glycosylation status on tumor and benign cells/MV was analyzed by western blotting (HG-EMP, highly glycosylated EMMPRIN; IG-EMP, intermediately glycosylated EMMPRIN; LG-EMP, lowly glycosylated EMMPRIN). (B) T-MVM were deglycosylated by PNGaseF (visible as an unspecific band in lanes 1 and 3) under denaturing or nondenaturing conditions and the EMMPRIN glycosylation status was assessed by western blotting. (C) Microinvasion assay of MCF-7 cells stimulated with deglycosylated T-MV (10 µg/ml, non-denat. conditions) (mean ± SD, n = 3, *P < 0.001).
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4401212&req=5

MJU047F4: High glycosylation is essential for the proinvasive function of EMMPRIN. (A) The EMMPRIN glycosylation status on tumor and benign cells/MV was analyzed by western blotting (HG-EMP, highly glycosylated EMMPRIN; IG-EMP, intermediately glycosylated EMMPRIN; LG-EMP, lowly glycosylated EMMPRIN). (B) T-MVM were deglycosylated by PNGaseF (visible as an unspecific band in lanes 1 and 3) under denaturing or nondenaturing conditions and the EMMPRIN glycosylation status was assessed by western blotting. (C) Microinvasion assay of MCF-7 cells stimulated with deglycosylated T-MV (10 µg/ml, non-denat. conditions) (mean ± SD, n = 3, *P < 0.001).
Mentions: In order to test whether differential EMMPRIN expression could explain the proinvasive effect of T-MV in contrast to benign MV, we analyzed EMMPRIN expression in various cells and their MV. EMMPRIN was strongly enriched in T-MV compared with their parental cells, but only weakly expressed on platelet-derived MV (P-MV) (Figure 4A) that were previously identified as being not proinvasive (Menck et al., 2013). Surprisingly, the strongest EMMPRIN enrichment was demonstrated on MV from the benign hTERT-HME1. Analysis of the protein size revealed that all investigated MV contained a large form of EMMPRIN ranging from 45 to 65 kDa, indicating glycosylation at multiple sites. However, within this fraction, two different glycoforms can be distinguished: (i) a very highly glycosylated protein (HG-EMMPRIN, ∼50–65 kDa) characterizing all proinvasive MV, and (ii) a variant with intermediate glycosylation (IG-EMMPRIN, ∼45 kDa) in all non-proinvasive MV. The low glycosylation variant (LG-EMMPRIN, 32 kDa) was only very weakly detectable.Figure 4

Bottom Line: Uptake of T-MV is essential for the proinvasive effect.Interestingly, the effect of EMMPRIN-bearing MV is not mediated by matrix metalloproteinases but by activation of the p38/MAPK signaling pathway in the tumor cells.In conclusion, T-MV stimulate cancer cell invasion via a direct feedback mechanism dependent on highly glycosylated EMMPRIN.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology/Oncology, University Medical Center Göttingen, 37099 Göttingen, Germany.

No MeSH data available.


Related in: MedlinePlus