Abstract
Membrane-derived extracellular vesicles, referred to as microvesicles (MVs), have been proposed to participate in several cancer diseases. In this study, MV fractions were isolated by differential ultracentrifugation from a metastatic breast cancer (BC) cell line MDA-MB-231 and a non-cancerous breast cell line MCF10A, then analyzed by nano-liquid chromatography coupled to tandem mass spectrometry. A total of 1519 MV proteins were identified from both cell lines. The data obtained were compared to previously analyzed proteins from small extracellular vesicles (sEVs), revealing 1272 proteins present in both MVs and sEVs derived from the MDA-MB-231 cell line. Among the 89 proteins unique to MDA-MB-231 MVs, three enzymes: ornithine aminotransferase (OAT), transaldolase (TALDO1) and bleomycin hydrolase (BLMH) were previously proposed as cancer therapy targets. These proteins were enzymatically validated in cells, sEVs, and MVs derived from both cell lines. The specific activity of OAT and TALDO1 was significantly higher in MDA-MB-231-derived MVs than in MCF10A MVs. BLMH was highly expressed in MDA-MB-231-derived MVs, compared to MCF10A MVs. This study shows that MVs carry functional metabolic enzymes and provides a framework for future studies of their biological role in BC and potential in therapeutic applications.
Highlights
Breast cancer (BC) continues to be one of the leading causes of death for women worldwide, accounting for 25% of all cancers [1]
The results show that the amount of bleomycin hydrolase (BLMH) in MDA-MB-231 cell-free extract (CFE) is higher in comparison to MCF10A CFE (Figure 7)
We utilized proteomicsEVs datatofrom we present proteomics of breast cell line-derived shedpreviously light on analyzed MDA-MB-231 and MCF10A cell line-derived small extracellular vesicles (sEVs) [30]
Summary
Breast cancer (BC) continues to be one of the leading causes of death for women worldwide, accounting for 25% of all cancers [1]. Overall mortality rates of BC are decreasing in developed countries due to increased awareness of self-examination procedures, improvement in early detection methods, and treatment advances [2,3]. The mortality rates of metastatic BC subtypes remain alarming, with an estimated five-year survival rate of 36% [4]. The development of metastatic BC is characterized by certain hallmarks—modulation of microenvironment, angiogenesis, sustained growth and tissue invasion—that rely on cell–cell signaling and communication [5,6]. Extracellular vesicles (EVs) have emerged as a novel mechanism of intercellular communication that does not require direct cellular contact [7]. Encapsulated by a phospholipid bilayer, these vesicles induce biologically relevant changes in recipient cells by transferring bioactive content (nucleic acids, proteins, and lipids) from donor to nearby or distant cells [8,9,10].
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