Journal: Technology in Cancer Research & Treatment
Article Title: Combined DNA-PK and PARP Inhibition as a Therapeutic Strategy in BRCA-Mutated Prostate Cancer: An in Vitro Pilot Study
doi: 10.1177/15330338251394948
Figure Lengend Snippet: BRCA1/2 Mutations and Deep Deletions Link to Poor Outcomes. (A-B) Frequency of BRCA1 (A) and BRCA2 (B) Alterations, Including Mutations (red) and Deep Deletions (Blue), Across the cBioPortal for Cancer Genomics Database for Prostate Cancer. The Patient-Weighted Average Alteration Frequency is Indicated by a Dashed Vertical Line. (C) Kaplan–Meier Analysis of Overall Survival Comparing Patients with BRCA1/2 Alterations (Red) to Those Without (Blue), Demonstrating Significantly Reduced Survival in the Altered Group. Δ Median Survival = 27.89 Months. n = 308 in BRCA1/2 Altered Group; n = 3717 in the Unaltered Group. (D) Schematic of CRISPR-Cas9-Mediated Knockout (KO) of BRCA1 or BRCA2 in Prostate Cancer Cell Lines LNCaP. (E) Representative Virtual Lanes of Simple Western Validation of BRCA1 and BRCA2 Knockout Efficiency. (F) Representative Lane View and Densitometric Quantification Normalized to Vinculin are Shown. Graphs Represent Mean ± SEM from Three-Four Independent Experiments.
Article Snippet: Primary antibodies were used at a 1:100 dilution and included mouse anti-Vinculin (MAB6896, Bio-Techne, Minneapolis, MN, USA), rabbit anti- BRCA2 (10741S, Cell Signaling Technology, Danvers, MA, USA), mouse anti-BRCA1 (NB100-598, Bio-Techne, Minneapolis, MN, USA), and rabbit anti- DNA PKcs (ab32566, Abcam, Cambridge, UK).
Techniques: CRISPR, Knock-Out, Simple Western, Biomarker Discovery
