Triple-negative breast cancer cells treated with photodynamic therapy express differential proteins.
•Calreticulin and HSP90 were identified as differential proteins in MDA-MB-231 cell line.
•Peptides obtained from differential proteins showed immunogenic properties in silico.
•Peptides induce macrophages activation and TNF cytokine release in vitro.
AbstractBackgroundPhotodynamic therapy (PDT) is used to treat tumors through selective cytotoxic effects. PDT induces damage-associated molecular patterns (DAMPs) expression, which can cause an immunogenic death cell (IDC). In this study we identified potential immunogenic epitopes generated by PDT on triple-negative breast cancer cell line (MDA-MB-231).
MethodsMDA-MB-231 cells were exposed to PDT using ALA (160 µg/mL)/630 nm at 8 J/cm2. Membrane proteins were extracted and separated by 2D PAGE. Proteins overexpressed were identified by LC-MS/MS and analyzed in silico through a peptide-HLA docking in order to identify the epitopes with more immunogenicity and antigenicity properties, as well as lower allergenicity and toxicity activity. The selected peptides were evaluated in response to macrophage activation and cytokine release by flow cytometry.
ResultsDifferential proteins were overexpressed in the cells treated with PDT. A group of 16 peptides were identified from them, established in a rigorous selection by measuring antigenicity, immunogenicity, allergenicity, and toxicity in silico. The final selection was based on molecular dynamics, where 2 peptides showed the highest stability regarding to the RMSD value. These peptides were obtained from the proteins calreticulin and HSP90. The cytokine analysis evidenced macrophage activation by the releasing of TNF.
ConclusionTwo peptides were identified from calreticulin and HSP90; proteins induced by PDT in MDA-MB-231 cells. Both epitopes showed immunogenic potential as a peptide-based vaccine for triple-negative breast cancer.
KeywordsPhotodynamic therapy
Calreticulin
HSP90
Peptide-based vaccine
Triple negative breast cancer
AbbreviationsDAMPsdamage-associated molecular patterns
IDCimmunogenic death cell
MAPspeptides in multiple antigenic peptide
TNBCtriple-negative breast cancer
CTLcytotoxic T lymphocytes
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