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Serglycin is implicated in the promotion of aggressive phenotype of breast cancer cells.

Korpetinou A, Skandalis SS, Moustakas A, Happonen KE, Tveit H, Prydz K, Labropoulou VT, Giannopoulou E, Kalofonos HP, Blom AM, Karamanos NK, Theocharis AD - PLoS ONE (2013)

Bottom Line: Serglycin exhibited a strong cytoplasmic staining in these cells, observable at the cell periphery in a thread of filaments near the cell membrane, but also in filopodia-like structures.Serglycin was purified from conditioned medium of MDA-MB-231 cells, and represented the major proteoglycan secreted by these cells, having a molecular size of ~ 250 kDa and carrying chondroitin sulfate side chains, mainly composed of 4-sulfated (~ 87%), 6-sulfated (~ 10%) and non-sulfated (~ 3%) disaccharides.Interestingly, over-expression of serglycin lacking the glycosaminoglycan attachment sites failed to promote these cellular functions, suggesting that glycanation of serglycin is a pre-requisite for its oncogenic properties.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Biochemistry, Department of Chemistry, University of Patras, Patras, Greece.

ABSTRACT
Serglycin is a proteoglycan expressed by some malignant cells. It promotes metastasis and protects some tumor cells from complement system attack. In the present study, we show for the first time the in situ expression of serglycin by breast cancer cells by immunohistochemistry in patients' material. Moreover, we demonstrate high expression and constitutive secretion of serglycin in the aggressive MDA-MB-231 breast cancer cell line. Serglycin exhibited a strong cytoplasmic staining in these cells, observable at the cell periphery in a thread of filaments near the cell membrane, but also in filopodia-like structures. Serglycin was purified from conditioned medium of MDA-MB-231 cells, and represented the major proteoglycan secreted by these cells, having a molecular size of ~ 250 kDa and carrying chondroitin sulfate side chains, mainly composed of 4-sulfated (~ 87%), 6-sulfated (~ 10%) and non-sulfated (~ 3%) disaccharides. Purified serglycin inhibited early steps of both the classical and the lectin pathways of complement by binding to C1q and mannose-binding lectin. Stable expression of serglycin in less aggressive MCF-7 breast cancer cells induced their proliferation, anchorage-independent growth, migration and invasion. Interestingly, over-expression of serglycin lacking the glycosaminoglycan attachment sites failed to promote these cellular functions, suggesting that glycanation of serglycin is a pre-requisite for its oncogenic properties. Our findings suggest that serglycin promotes a more aggressive cancer cell phenotype and may protect breast cancer cells from complement attack supporting their survival and expansion.

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Serglycin is highly expressed in aggressive breast cancer cell lines.(A) Expression of serglycin across the 51 individual breast cancer cell lines grouped in the basal A (red), basal B (grey) and luminal (blue) subgroups. (B) Detection of serglycin gene transcripts in low aggressive MCF-7 (luminal), medium aggressive MDA-MB-468 (basal A) and high aggressive MDA-MB-231 (basal B) breast cancer cells (bold and underlined in figure 2A) cultured in the presence or absence of serum by RT-PCR. The PCR products were analyzed on 1% agarose gels stained with GelRed. (C). Subcellular distribution of serglycin in breast cancer cells. Immunofluorescence staining was done for serglycin (green), nuclei (blue) and F-actin (red) in high aggressive MDA-MB-231 and low aggressive MCF-7 breast cancer cells after permeabilization. MCF-7 cells stained faintly for serglycin in the cytoplasm. Serglycin was found in the perinuclear regions and juxtanuclear area typical of the Golgi complex in MDA-MB-231 cells (arrowheads), and in the cytoplasm and at the cell periphery (arrows). Serglycin distribution was observed in vesicular (V) and vesiculotubular structures (VST) in the cytoplasm (insert). Bars, 25 µm.
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pone-0078157-g002: Serglycin is highly expressed in aggressive breast cancer cell lines.(A) Expression of serglycin across the 51 individual breast cancer cell lines grouped in the basal A (red), basal B (grey) and luminal (blue) subgroups. (B) Detection of serglycin gene transcripts in low aggressive MCF-7 (luminal), medium aggressive MDA-MB-468 (basal A) and high aggressive MDA-MB-231 (basal B) breast cancer cells (bold and underlined in figure 2A) cultured in the presence or absence of serum by RT-PCR. The PCR products were analyzed on 1% agarose gels stained with GelRed. (C). Subcellular distribution of serglycin in breast cancer cells. Immunofluorescence staining was done for serglycin (green), nuclei (blue) and F-actin (red) in high aggressive MDA-MB-231 and low aggressive MCF-7 breast cancer cells after permeabilization. MCF-7 cells stained faintly for serglycin in the cytoplasm. Serglycin was found in the perinuclear regions and juxtanuclear area typical of the Golgi complex in MDA-MB-231 cells (arrowheads), and in the cytoplasm and at the cell periphery (arrows). Serglycin distribution was observed in vesicular (V) and vesiculotubular structures (VST) in the cytoplasm (insert). Bars, 25 µm.

Mentions: The expression of serglycin across 51 human breast cancer cell lines [21] clustered into three subgroups (Luminal, Basal A and Basal B) was obtained by the Cell Lines module in the Gene Set Analysis using GOBO database (http://co.bmc.lu.se/gobo/gobo.pl). Serglycin was over-expressed by breast cancer cells clustered into Basal B subgroup (Fig. 2A). Breast cancer cells that belong to Basal B appeared less differentiated, had a more mesenchymal-like appearance and were highly invasive, as compared to cells clustered into Basal A and Luminal subgroups [21]. To confirm these data in our cell lines, we examined the expression of serglycin in breast cancer cell lines that clustered into three subgroups (MCF-7, MDA-MB-468 and MDA-MB-231 cells, clustered to Luminal, Basal A and Basal B, respectively). Total RNAs from MDA-MB-231 (high aggressive), MDA-MB-468 (medium aggressive) and MCF-7 (low aggressive) breast cancer cells cultured in serum-free medium or in complete culture medium were reverse transcribed and amplified using specific primers for serglycin (Fig. 2B). It was found that high aggressive MDA-MB-231 cells expressed much higher levels of mRNA coding for serglycin compared to medium aggressive MDA-MB-468 cells (Fig. 2B), whereas low aggressive MCF-7 cells hardly expressed detectable amounts of mRNA for serglycin (Fig. 2B). In all cases, the cells cultured in complete medium showed a tendency to higher expression of the serglycin gene compared to cells cultured in serum-free culture medium, although this increase was not statistically significant (not shown).


Serglycin is implicated in the promotion of aggressive phenotype of breast cancer cells.

Korpetinou A, Skandalis SS, Moustakas A, Happonen KE, Tveit H, Prydz K, Labropoulou VT, Giannopoulou E, Kalofonos HP, Blom AM, Karamanos NK, Theocharis AD - PLoS ONE (2013)

Serglycin is highly expressed in aggressive breast cancer cell lines.(A) Expression of serglycin across the 51 individual breast cancer cell lines grouped in the basal A (red), basal B (grey) and luminal (blue) subgroups. (B) Detection of serglycin gene transcripts in low aggressive MCF-7 (luminal), medium aggressive MDA-MB-468 (basal A) and high aggressive MDA-MB-231 (basal B) breast cancer cells (bold and underlined in figure 2A) cultured in the presence or absence of serum by RT-PCR. The PCR products were analyzed on 1% agarose gels stained with GelRed. (C). Subcellular distribution of serglycin in breast cancer cells. Immunofluorescence staining was done for serglycin (green), nuclei (blue) and F-actin (red) in high aggressive MDA-MB-231 and low aggressive MCF-7 breast cancer cells after permeabilization. MCF-7 cells stained faintly for serglycin in the cytoplasm. Serglycin was found in the perinuclear regions and juxtanuclear area typical of the Golgi complex in MDA-MB-231 cells (arrowheads), and in the cytoplasm and at the cell periphery (arrows). Serglycin distribution was observed in vesicular (V) and vesiculotubular structures (VST) in the cytoplasm (insert). Bars, 25 µm.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3815026&req=5

pone-0078157-g002: Serglycin is highly expressed in aggressive breast cancer cell lines.(A) Expression of serglycin across the 51 individual breast cancer cell lines grouped in the basal A (red), basal B (grey) and luminal (blue) subgroups. (B) Detection of serglycin gene transcripts in low aggressive MCF-7 (luminal), medium aggressive MDA-MB-468 (basal A) and high aggressive MDA-MB-231 (basal B) breast cancer cells (bold and underlined in figure 2A) cultured in the presence or absence of serum by RT-PCR. The PCR products were analyzed on 1% agarose gels stained with GelRed. (C). Subcellular distribution of serglycin in breast cancer cells. Immunofluorescence staining was done for serglycin (green), nuclei (blue) and F-actin (red) in high aggressive MDA-MB-231 and low aggressive MCF-7 breast cancer cells after permeabilization. MCF-7 cells stained faintly for serglycin in the cytoplasm. Serglycin was found in the perinuclear regions and juxtanuclear area typical of the Golgi complex in MDA-MB-231 cells (arrowheads), and in the cytoplasm and at the cell periphery (arrows). Serglycin distribution was observed in vesicular (V) and vesiculotubular structures (VST) in the cytoplasm (insert). Bars, 25 µm.
Mentions: The expression of serglycin across 51 human breast cancer cell lines [21] clustered into three subgroups (Luminal, Basal A and Basal B) was obtained by the Cell Lines module in the Gene Set Analysis using GOBO database (http://co.bmc.lu.se/gobo/gobo.pl). Serglycin was over-expressed by breast cancer cells clustered into Basal B subgroup (Fig. 2A). Breast cancer cells that belong to Basal B appeared less differentiated, had a more mesenchymal-like appearance and were highly invasive, as compared to cells clustered into Basal A and Luminal subgroups [21]. To confirm these data in our cell lines, we examined the expression of serglycin in breast cancer cell lines that clustered into three subgroups (MCF-7, MDA-MB-468 and MDA-MB-231 cells, clustered to Luminal, Basal A and Basal B, respectively). Total RNAs from MDA-MB-231 (high aggressive), MDA-MB-468 (medium aggressive) and MCF-7 (low aggressive) breast cancer cells cultured in serum-free medium or in complete culture medium were reverse transcribed and amplified using specific primers for serglycin (Fig. 2B). It was found that high aggressive MDA-MB-231 cells expressed much higher levels of mRNA coding for serglycin compared to medium aggressive MDA-MB-468 cells (Fig. 2B), whereas low aggressive MCF-7 cells hardly expressed detectable amounts of mRNA for serglycin (Fig. 2B). In all cases, the cells cultured in complete medium showed a tendency to higher expression of the serglycin gene compared to cells cultured in serum-free culture medium, although this increase was not statistically significant (not shown).

Bottom Line: Serglycin exhibited a strong cytoplasmic staining in these cells, observable at the cell periphery in a thread of filaments near the cell membrane, but also in filopodia-like structures.Serglycin was purified from conditioned medium of MDA-MB-231 cells, and represented the major proteoglycan secreted by these cells, having a molecular size of ~ 250 kDa and carrying chondroitin sulfate side chains, mainly composed of 4-sulfated (~ 87%), 6-sulfated (~ 10%) and non-sulfated (~ 3%) disaccharides.Interestingly, over-expression of serglycin lacking the glycosaminoglycan attachment sites failed to promote these cellular functions, suggesting that glycanation of serglycin is a pre-requisite for its oncogenic properties.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Biochemistry, Department of Chemistry, University of Patras, Patras, Greece.

ABSTRACT
Serglycin is a proteoglycan expressed by some malignant cells. It promotes metastasis and protects some tumor cells from complement system attack. In the present study, we show for the first time the in situ expression of serglycin by breast cancer cells by immunohistochemistry in patients' material. Moreover, we demonstrate high expression and constitutive secretion of serglycin in the aggressive MDA-MB-231 breast cancer cell line. Serglycin exhibited a strong cytoplasmic staining in these cells, observable at the cell periphery in a thread of filaments near the cell membrane, but also in filopodia-like structures. Serglycin was purified from conditioned medium of MDA-MB-231 cells, and represented the major proteoglycan secreted by these cells, having a molecular size of ~ 250 kDa and carrying chondroitin sulfate side chains, mainly composed of 4-sulfated (~ 87%), 6-sulfated (~ 10%) and non-sulfated (~ 3%) disaccharides. Purified serglycin inhibited early steps of both the classical and the lectin pathways of complement by binding to C1q and mannose-binding lectin. Stable expression of serglycin in less aggressive MCF-7 breast cancer cells induced their proliferation, anchorage-independent growth, migration and invasion. Interestingly, over-expression of serglycin lacking the glycosaminoglycan attachment sites failed to promote these cellular functions, suggesting that glycanation of serglycin is a pre-requisite for its oncogenic properties. Our findings suggest that serglycin promotes a more aggressive cancer cell phenotype and may protect breast cancer cells from complement attack supporting their survival and expansion.

Show MeSH
Related in: MedlinePlus