Novel Processed Form of Syndecan-1 Shed from SCC-9 Cells Plays a Role in Cell Migration

Abstract

The extracellular milieu is comprised in part by products of cellular secretion and cell surface shedding. The presence of such molecules of the sheddome and secretome in the context of the extracellular milieu may have important clinical implications. In cancer they have been hypothesized to play a role in tumor growth and metastasis. The objective of this study was to evaluate whether the sheddome/secretome from two cell lines could be correlated with their potential for tumor development. Two epithelial cell lines, HaCaT and SCC-9, were chosen based on their differing abilities to form tumors in animal models of tumorigenesis. These cell lines when stimulated with phorbol-ester (PMA) showed different characteristics as assessed by cell migration, adhesion and higher gelatinase activity. Proteomic analysis of the media from these treated cells identified interesting, functionally relevant differences in their sheddome/secretome. Among the shed proteins, soluble syndecan-1 was found only in media from stimulated tumorigenic cells (SCC-9) and its fragments were observed in higher amount in the stimulated tumorigenic cells than stimulated non-tumorigenic cells (HaCaT). The increase in soluble syndecan-1 was associated with a decrease in membrane-bound syndecan-1 of SCC-9 cells after PMA stimuli. To support a functional role for soluble syndecan-1 fragments we demonstrated that the synthetic syndecan-1 peptide was able to induce cell migration in both cell lines. Taken together, these results suggested that PMA stimulation alters the sheddome/secretome of the tumorigenic cell line SCC-9 and one such component, the syndecan-1 peptide identified in this study, was revealed to promote migration in these epithelial cell lines.

Acidulated Phosphate Fluoride Application Changes the Protein Composition of Human Acquired Enamel Pellicle

“…( table 1 ). Considering the fold change of 1.5 [Aragão et al, 2012a;Paes Leme et al, 2012], amylase has similar abundance among the samples and it was identified in a notable manner with the highest spectral counts, which means a high abundance of this protein in the acquired enamel pellicle independently of the enamel pretreatment. This result is consistent with earlier findings that indicated that amylase is one of the major components of pellicle [Hannig et al, 2004[Hannig et al, , 2005Siqueira et al, 2012b].…”

“…All datasets of proteins were processed using the workflow feature in Proteome Discoverer software v. 1.3 and the msf files were analyzed in ScaffoldQ+ v. 3.3.2 (Proteome Software), filtered using Xcorr cutoffs (+1 > 1.8, +2 > 2.2, +3 > 2.5 and +4 > 3.5), using the normalized number of total spectra counting to evaluate the protein abundance [Didangelos et al, 2011;Aragão et al, 2012a]. Spectral count normalizing entails averaging the spectrum counts across the samples, and then multiplying the spectrum counts in each sample by the average divided by the individual sample sum.…”

The Penicillium echinulatum Secretome on Sugar Cane Bagasse

“…and searched against the NCBI Database (11346708 sequences; 3868708671 residues; Version 2011; Fungi Taxonomy:678749 sequences) using the MASCOT v.2.3.01 (Matrix Science Ltd.) search engine with carbamidomethylation as a fixed modification, oxidation of methionine as a variable modification, one trypsin missed cleavage and a tolerance of 0.1 Da for both precursors and fragment ions. Only peptides with a minimum of six amino acid residues that showed significant threshold (p<0.05) in Mascot-based score were considered in the results [26]. The false positive identification was minimized using a cutoff corresponding to a protein confidence level of 95% (FDR of <1.0%).…”

Reduced serum levels of syndecan‐1 in patients with tongue squamous cell carcinoma

“…Defective expression of syndecan‐1 in mice results in defects in wound healing . The decrease of syndecan‐1 expression on tumor cells is associated with the release of the molecule from the cell surface as well as cytoplasmic or nuclear sequestration . Syndecan‐1 can be cleaved from the cell surface by the action of matrix metalloproteinases under the influence of chemokines .…”

“…The downregulation of syndecan‐1 is shown to be associated with the higher risk of metastasis and unfavorable outcome in head and neck SCC . The decrease of syndecan‐1 expression on tumor cells is shown to be associated with the release of the molecule from the cell surface as well as cytoplasmic or nuclear sequestration . In head and neck SCC, the decrease in the expression of the syndecan‐1 molecule is reported to be concomitant with the induction of its expression in the stromal cells of the tumor, which correlates with the recurrence and tumor‐specific death in 2 years follow‐up …”