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Induction of Apoptosis by Peptide J18 in Ovarian Cancer Cells
Author(s) -
Oldenburg Kimberly,
Wieskamp Matthew,
Soendergaard Mette
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.782.19
Subject(s) - ovarian cancer , apoptosis , peptide , dimethyl sulfoxide , chemistry , microbiology and biotechnology , cytotoxic t cell , gentamicin , fetal bovine serum , cancer cell , andrology , medicine , pharmacology , cell , cancer , biology , biochemistry , in vitro , antibiotics , organic chemistry
Ovarian cancer is the fifth leading cause of cancer‐related deaths for women, which is correlated to a low rate of early‐stage detection. Currently, standard treatment involves cytoreductive surgery and chemotherapy. However, patients often present with recurrent and resistant disease leading to overall low 5‐year survival rates of approximately 40%. Bacteriophage (phage) display technology was previously used to identify peptide J18 that binds specifically to human ovarian adenocarcinoma (SKOV‐3) cells. In addition, J18 was shown to reduce cell viability via the 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) reduction assay. In order to further investigate cytotoxic effects of peptide J18, induction of apoptosis was measured by a caspase 3/7 assay. Briefly, SKOV‐3 cells were grown to 90% confluency on 96‐well plates in McCoy's 5A cell medium supplemented with 10% fetal bovine serum and 50 μg/mL gentamicin at 37°C and 5% CO2. Cells were then treated with 0.25 mg/mL peptide J18, 0.25 mg/mL peptide N35 (negative control), 100 nM paclitaxel, or dimethyl sulfoxide (DMSO; vehicle) for 12 h, after which 100 μL Apo‐ONE Caspase‐3/7 Reagent (Promega, WI) was added and incubated at 37°C for 3 h. The activity of caspase 3/7 was then measured fluorescently (λex=499 nm, λem=521 nm) using a microplate reader (SpectraMax Gemini EM, Molecular Devices, CA). The relative fluorescent units (RFU) was measured to be 409.7±96.0, 560.6±84.1, 652.5±149.1, and 648.0±47.5 (mean±standard deviation) for DMSO, N35, J18, and paclitaxel, respectively. Results showed that J18 and paclitaxel significantly (p<0.05) increased caspase 3/7 activity compared to DMSO (control), while N35 (negative control) was unchanged. Further, there was no significant difference (p>0.5) between J18 and paclitaxel at the concentrations measured. These results indicate that J18 reduces cell viability in human ovarian adenocarcinoma cells by apoptosis, and that the peptide may be utilized in the treatment of the disease. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .