Therefore, inhibition of BRCA2 may be a useful strategy to decrease metastatic burden in patients treated with cisplatin. Kif15-IN-2 Funding This work is supported by the Ontario Center of Excellence C2C award #20159. Conflict of interest The authors have no conflict of interest to declare. Supporting information The following are the supplementary data related to this article: Supplementary Physique?1 BRCA2 knockdown modulates cisplatin\ or melphalan\induced A549 cell proliferation. ASO using a Student’s t\test (p?Rabbit polyclonal to BMPR2 than cells pre\treated with control ASO. Furthermore, in BRCA2 ASO\treated cells, the respiration decrease was evident 10?h after addition of cisplatin and 15?h earlier than in cells treated with control ASO (Figure?5a). Respiration began to decrease in response to cisplatin in BRCA2\treated cells 10?h prior to observable reduction in adhesion, suggesting that respiration reduction occurred independent of changes in cell number or viability (Physique?5b). However, no difference in acidification (a?measure of glycolysis) was observed between the BRCA2 ASO and control ASO Kif15-IN-2 groups treated with cisplatin (Physique?5c). Open in a separate window Physique 5 BRCA2 modulates tumor cell metabolic response following cisplatin treatment. A549 cells were exposed to cisplatin (6?M, 24?h) following 6?h of incubation in medium to determine baseline metabolic levels. At 24?h after addition of cisplatin, medium was exchanged for medium without cisplatin. Measurements of oxygen consumption (a), impedance (b), and changes in medium pH (c) were conducted. Magenta = Control ASO, Blue = BRCA2 ASO, Green = Control ASO + Cisplatin, Red = BRCA2 ASO + cisplatin. A549 cells were transfected with control or BRCA2 ASO and then treated with cisplatin (6?M). Mitotracker staining (d) and glucose uptake (e) were determined using flow cytometry. Unstained control = Red, Control ASO = blue, BRCA2 ASO = orange, Control ASO + Cisplatin = light green, BRCA2 ASO + cisplatin = dark green. *Different from cells treated with control ASO using a Student’s t\test (p?