title: Topotecan synergizes with CHEK1 (CHK1) inhibitor to induce apoptosis in ovarian cancer cells.
abstract: BACKGROUND
  Topotecan (TPT) is a therapeutic option for women with platinum-resistant or -refractory ovarian cancer. However, the dose-limiting toxicity of TPT is myelosuppression. This led us to seek a combination treatment to augment TPT anti-cancer activity in a cancer-targeted manner. Ovarian serous cancers, a major subtype, show dysregulated DNA repair pathway and often display a high level of CHEK1 (CHK1), a cell cycle regulator and DNA damage sensor. CHEK1 inhibitors are a novel approach to treatment, and have been used as single agents or in combination chemotherapy in many cancers.
  
  
  METHODS
  We evaluated the cellular effects of TPT in a panel of high grade serous (HGS) and non-HGS ovarian cancer cells. We then determined IC50s of TPT in the absence and presence of CHEK1 inhibitor, PF477736. Synergism between TPT and PF477736 was calculated based on cellular viability assays. Cytotoxic effect of the combined treatment was compared with apoptotic activities by Caspase3/7 activity assay and Western blotting of cleaved-PARP1 and γH2AX.
  
  
  RESULTS
  Non-HGS ovarian cancer cells were generally more sensitive to TPT treatment compared to HGS ovarian cancer cells. When combined with CHEK1 inhibitor, TPT potently and synergistically inhibited the proliferation of HGS ovarian cancer cells. This dramatic synergism in cellular toxicity was consistent with increases in markers of apoptosis.
  
  
  CONCLUSIONS
  Our findings suggest that the addition of CHEK1 inhibitor increases the response of ovarian cancer cells to TPT. Furthermore, reduced dosages of both drugs achieved maximal cytotoxic effects by combining TPT with CHEK1 inhibitor. This strategy would potentially minimize side effects of the drugs for extended clinical benefit.
authors.affiliation: Women's Malignancies Branch, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD, 20892, USA. marianne.kim@nih.gov.
authors.forename: Marianne K
authors.initials: MK
authors.lastname: Kim
authors.affiliation: Women's Malignancies Branch, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD, 20892, USA. jana.james@nih.gov.
authors.forename: Jana
authors.initials: J
authors.lastname: James
authors.affiliation: Women's Malignancies Branch, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD, 20892, USA. annunzic@mail.nih.gov.
authors.forename: Christina M
authors.initials: CM
authors.lastname: Annunziata
mesh_terms: D000971:Antineoplastic Combined Chemotherapy Protocols; D017209:Apoptosis / Q000187:drug effects; D001570:Benzodiazepinones / Q000008:administration & dosage*; D000077216:Carcinoma, Ovarian Epithelial; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D000071877:Checkpoint Kinase 1; D002945:Cisplatin / Q000008:administration & dosage / Q000009:adverse effects; D004249:DNA Damage / Q000187:drug effects; D004260:DNA Repair / Q000187:drug effects; D004357:Drug Synergism; D005260:Female; D006801:Humans; D009375:Neoplasms, Glandular and Epithelial / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D010051:Ovarian Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D011494:Protein Kinases / Q000235:genetics*; D011720:Pyrazoles / Q000008:administration & dosage*; D059004:Topoisomerase I Inhibitors / Q000008:administration & dosage* / Q000009:adverse effects; D019772:Topotecan / Q000008:administration & dosage* / Q000009:adverse effects
chemical_list: D001570:Benzodiazepinones; C532366:PF 00477736; D011720:Pyrazoles; D059004:Topoisomerase I Inhibitors; D019772:Topotecan; D011494:Protein Kinases; C000605867:CHEK1 protein, human; D000071877:Checkpoint Kinase 1; D002945:Cisplatin