We focused on the proteomic characterization of nonmineralizing (NMOBs) and mineralizing (MOBs) human osteoblast (SV-HFOs) EVs and investigated their effect on human prostate cancer (PC3) cells by microscopic, proteomic, and gene expression analyses.
Proteomic analysis showed that 97% of the proteins were shared among NMOB and MOB EVs, and 30% were novel osteoblast-specific EV proteins.
Cysteine-rich protein 61 (Cyr61) was found to be abundantly present in MSCs.
The presence of Cyr61 was confirmed by immunofluorescence staining and immunoblot analysis.
Importantly, addition of recombinant Cyr61 polypeptides restores the angiogenic activity of Cyr61-depleted secretome.
Of these, approximately 90% have somatic mutations in cancer, 20% bear germline mutations that predispose to cancer and 10% show both somatic and germline mutations.
A list of genes strongly implicated in cancer (n=555) has been defined through the cancer Gene Census, catalogue of somatic mutations in cancer (COSMIC). List of different genetic alterations in 555 genes that are implicated in cancer.
This is followed by isotopic labeling, which allows for multiplex comparisons, pre-fractionated and subjected to LC-MS/MS using a state of the art Mass Spectrometer (Thermo Q Exactive Plus), followed by data analysis.
Therefore, we recently devised a method for isolation of interstitial fluid from cancers that is suitable for isotope-labeling with subsequent MS-based quantification and identification of secreted and shed proteins (Gomez-Auli et al. After depletion of highly abundant proteins, for better proteome coverage, proteins are precipitated and digested.