Exponentially growing A2780 cells are plated in 96 well microtitre plates. For experiments studying the effect of sequence of exposure to 17-AAG or Carboplatin, cells are exposed to increasing concentrations of 17-AAG or Carboplatin for 24 h. A period of 24-h exposure to the first agent is chosen so that the A2780 cells will be exposed to the first drug for at least one doubling time (18-24 h). The cells are then washed with sterile phosphate buffered saline and the medium is replenished. Following this, the second drug (to which the cells are not exposed to in the first 24 h) or medium is added for 96 h. SRB assays are carried out. All experiments are carried out in triplicate^[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Mice^[2]
The A2780 human ovarian cancer cell line is grown as a subcutaneous xenograft in female athymic NCr nude mice (nu/nu) by injecting 4 × 10⁶ cells in each flank. Mice with established tumors corresponding to a mean volume of 0.69 mm³ are randomized into groups (six animals each) for treatment with either control vehicle (43% ethanol, 33% polypropylene glycol and 24% cremaphor diluted 1:7 with sterile water) days 1-4, 17-AAG (80 mg/kg intraperitonially, days 1-4), Carboplatin (60 mg/kg IP day 0) or a combination of 17-AAG (80 mg/kg IP days 1-4) and Carboplatin (60 mg/kg IP day 0). Tumor growth is assessed three times weekly and tumor volumes are calculated according to a validated formula: volume = 4.19 × (a/4 + b/4)³, where a is the longer and b the shorter diameter. Tumor volumes are then expressed as a proportion of the volume at the start of treatment (relative tumor volume)^[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

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  [2]. Banerji U, et al. An in vitro and in vivo study of the combination of the heat shock protein inhibitor 17-allylamino-17-demethoxygeldanamycin and carboplatin in human ovarian cancer models. Cancer Chemother Pharmacol. 2008 Oct;62(5):769-78.  [Content Brief]

  [3]. Clark CC, et al. Enhancement of synthetic lethality via combinations of ABT-888, a PARP inhibitor, and carboplatin in vitro and in vivo using BRCA1 and BRCA2 isogenic models. Mol Cancer Ther. 2012 Sep;11(9):1948-58.  [Content Brief]

  [4]. Dela Cruz FS, et al. A case study of an integrative genomic and experimental therapeutic approach for rare tumors: identification of vulnerabilities in a pediatric poorly differentiated carcinoma. Genome Med. 2016 Oct 31;8(1):116.  [Content Brief]

  [5]. Hall MD, Telma KA, Chang KE, et al. Say no to DMSO: dimethylsulfoxide inactivates cisplatin, carboplatin, and other platinum complexes. Cancer Res. 2014;74(14):3913-3922.  [Content Brief]