Division of Biosciences, College of Health, Medicine and Life Sciences, Brunel University London, Uxbridge UB8 3PH, UK.
Mount Vernon Cancer Centre, Rickmansworth Road, Northwood HA6 2RN, UK.
Cells. 2021 Sep 15;10(9):2434. doi: 10.3390/cells10092434.
Approximately 50% of ovarian cancer patients harbour homologous recombination repair deficiencies. These deficiencies have been successfully targeted using poly (ADP-ribose) polymerase inhibitors (PARPi) particularly for patients harbouring BRCA1/2 mutations. The aim of this study is to assess the effects of the PARPi rucaparib in vitro using cell lines with BRCA2 mutations in comparison to those with BRCA2 wild type.
Cell proliferation assays, RT-qPCR, immunofluorescence, annexin V/PI assays were used to assess the effects of rucaparib in vitro.
The BRCA2 mutant ovarian cancer cell line PEO1 exhibited higher PARP1 activity when treated with HO compared to wild type cell lines. The migratory and proliferative capacity of PEO1 cells was compromised following treatment with rucaparib 10 µM compared to BRCA2 wild-type cell lines via a mechanism involving the mTOR pathway. Rucaparib treatment significantly increased DNA damage primarily in PEO1 cells and SKOV3 cells compared with wild type.
Appropriate identification of robust predictive biomarkers for homologous recombination deficiency using 'liquid' biopsies would facilitate the identification of patients suitable for PARPi therapy. Preliminary efforts to undertake such testing are described here. This study also demonstrates the mechanisms of action of rucaparib (PARPi) which may involve elements of the mTOR pathway.
约 50%的卵巢癌患者存在同源重组修复缺陷。这些缺陷已成功地通过聚（ADP-核糖）聚合酶抑制剂（PARPi）进行靶向治疗，特别是针对携带 BRCA1/2 突变的患者。本研究旨在评估 PARPi 鲁卡帕尼在体外对携带 BRCA2 突变的细胞系的影响，并与携带 BRCA2 野生型的细胞系进行比较。
采用细胞增殖试验、RT-qPCR、免疫荧光、膜联蛋白 V/PI 法评估鲁卡帕尼在体外的作用。
与野生型细胞系相比，BRCA2 突变卵巢癌细胞系 PEO1 在用 HO 处理时表现出更高的 PARP1 活性。与 BRCA2 野生型细胞系相比，鲁卡帕尼 10μM 处理后，PEO1 细胞的迁移和增殖能力受损，这一机制涉及 mTOR 通路。与野生型相比，鲁卡帕尼处理后，PEO1 细胞和 SKOV3 细胞的 DNA 损伤明显增加。
通过“液体”活检适当识别同源重组缺陷的稳健预测生物标志物，将有助于确定适合 PARPi 治疗的患者。这里描述了初步进行此类检测的努力。本研究还证明了鲁卡帕尼（PARPi）的作用机制，可能涉及 mTOR 通路的某些元素。
