Label‐Free Quantitative Phosphoproteomics Reveals Regulation of Vasodilator‐Stimulated Phosphoprotein upon Stathmin‐1 Silencing in a Pair of Isogenic Colorectal Cancer Cell Lines

Abstract

In this communication, we present the phosphoproteome changes in an isogenic pair of colorectal cancer cell lines, viz., the poorly metastatic HCT-116 and the highly metastatic derivative E1, upon stathmin-1 (STMN1) knockdown. The aim was to better understand how the alterations of the phosphoproteins in these cells are involved in cancer metastasis. After the phosphopeptides were enriched using the TiO HAMMOC approach, comparative proteomics analysis was carried out using sequential window acquisition of all theoretical mass spectra-MS. Following bioinformatics analysis using MarkerView and OneOmics platforms, we identified a list of regulated phosphoproteins that may play a potential role in signaling, maintenance of cytoskeletal structure, and focal adhesion. Among these phosphoproteins, was the actin cytoskeleton regulator protein, vasodilator-stimulated phosphoprotein (VASP), where its change in phosphorylation status was found to be concomitant with STMN1-associated roles in metastasis. We further showed that silencing of stathmin-1 altered the expression, subcellular localization and phosphorylation status of VASP, which suggested that it might be associated with remodeling of the cell cytoskeleton in colorectal cancer metastasis.

Proteomic investigation on bio-corona of Au, Ag and Fe nanoparticles for the discovery of triple negative breast cancer serum protein biomarkers

“…The export will generate three files containing quantitative information about individual ions, the summed intensity of different ions for a particular peptide, and the summed intensity of different peptides for a specific protein. MarkerView has been used for the analysis of SWATH-MS data reported in other proteomics studies [39,40] because of its dataindependent method of quantitation. MarkerView uses processing algorithms that accurately find chromatographic and spectral peaks direct from the raw SWATH data.…”

Proteomic Analysis in Morquio A Cells Treated with Immobilized Enzymatic Replacement Therapy on Nanostructured Lipid Systems

“…The export generated three files containing quantitative information about individual ions, the summed intensity of the different ions for a particular peptide, and the summed intensity of different peptides for a particular protein. Marker View was used for analysis of the SWATH-MS data reported in other proteomics studies [34,62,63,64] because of its data-independent method of quantization. Marker View uses processing algorithms that accurately find chromatographic and spectral peaks direct from the raw SWATH-MS data.…”

“…Therefore, proteomics provides important biological information that contributes to the advance in our understanding of pathophysiological mechanisms and aid in the identification of novel biomarkers for different diseases [30,32,33]. Specific biomarkers identified from proteomics may be useful in diagnosis, prognosis, and as evaluators of treatment outcomes [34,35,36,37,38,39,40,41,42].…”

Exosomes in esophageal cancer: A review on tumorigenesis, diagnosis and therapeutic potential

“…Stathmin-1: Stathmin-1 is a microtubule-destabilizing cytosolic phosphoprotein that plays an important role in tumor cell proliferation and migration and may be regulated by miR-34a[30,31], miR-223[32] and miR-193b[33]. It is associated with cancer metastasis and poor prognosis in osteosarcoma[31,34], prostate cancer[35,36], head and neck squamous cell carcinoma[37], hepatocellular carcinoma[32], colorectal cancer[33,38], gallbladder carcinoma[39] and non-small-cell lung cancer[40]. In esophageal squamous cell carcinoma (ESCC), stathmin-1 is also vital for ESCC invasiveness and predicts a poor prognosis[41].…”