Department of Biochemistry and Biophysics, School of Medicine, University of North Carolina, CB #7260, Genetic Medicine Building, Chapel Hill, NC, 27599-7260, USA.
Cancer Chemother Pharmacol. 2012 Mar;69(3):665-77. doi: 10.1007/s00280-011-1744-3. Epub 2011 Oct 4.
To characterize the cellular action mechanism of Debio 0507, we compared the major DNA adducts formed by Debio 0507- and oxaliplatin-treated HCT116 human colon carcinoma cells by a combination of inductively coupled plasma mass spectrometry (ICP-MS) and ultraperformance liquid chromatography mass spectrometry (UPLC-MS/MS).
HCT116 cells were treated with IC(50) doses of Debio 0507 or oxaliplatin for 3 days. Total cellular Pt-DNA adducts were determined by ICP-MS. The DNA was digested, and the major Pt-DNA adducts formed by both drugs were characterized by UPLC/MS/MS essentially as described previously for cisplatin (Baskerville-Abraham et al. in Chem Res Toxicol 22:905-912, 2009).
The Pt level/deoxynucleotide was 7.4/10(4) for DNA from Debio 0507-treated cells and 5.5/10(4) for oxaliplatin-treated cells following a 3-day treatment at the IC(50) for each drug. UPLC-MS/MS in the positive ion mode confirmed the major Pt-DNA adducts formed by both drugs were dach-Pt-d(GpG) (904.2 m/z → 610 m/z and 904.2 m/z → 459 m/z) and dach-Pt-d(ApG) (888.2 m/z → 594 m/z and 888.2 m/z → 459 m/z).
These data show that the major DNA adducts formed by Debio 0507 are the dach-Pt-d(GpG) and dach-Pt-d(ApG) adducts and at equitoxic doses Debio 0507 and oxaliplatin form similar levels of dach-Pt-d(GpG) and dach-Pt-d(ApG) adducts. This suggests that the action mechanisms of Debio 0507 and oxaliplatin are similar at a cellular level.
为了研究 Debio 0507 的细胞作用机制，我们通过电感耦合等离子体质谱（ICP-MS）和超高效液相色谱-质谱联用（UPLC-MS/MS）相结合的方法比较了 Debio 0507 和奥沙利铂处理的 HCT116 人结肠癌细胞中形成的主要 DNA 加合物。
用 Debio 0507 或奥沙利铂的 IC50 剂量处理 HCT116 细胞 3 天。通过 ICP-MS 测定总细胞内 Pt-DNA 加合物。对 DNA 进行消化，并用 UPLC/MS/MS 对两种药物形成的主要 Pt-DNA 加合物进行了表征，其方法与先前顺铂（Baskerville-Abraham 等人，Chem Res Toxicol 22:905-912, 2009）的方法基本相同。
在每种药物的 IC50 处理 3 天后，Debio 0507 处理组细胞的 Pt/脱氧核苷酸水平为 7.4/10(4)，奥沙利铂处理组细胞的 Pt/脱氧核苷酸水平为 5.5/10(4)。正离子模式下的 UPLC-MS/MS 证实了两种药物形成的主要 Pt-DNA 加合物均为 dach-Pt-d(GpG)（904.2 m/z→610 m/z 和 904.2 m/z→459 m/z）和 dach-Pt-d(ApG)（888.2 m/z→594 m/z 和 888.2 m/z→459 m/z）。
这些数据表明，Debio 0507 形成的主要 DNA 加合物为 dach-Pt-d(GpG)和 dach-Pt-d(ApG)加合物，在等毒性剂量下，Debio 0507 和奥沙利铂形成相似水平的 dach-Pt-d(GpG)和 dach-Pt-d(ApG)加合物。这表明 Debio 0507 和奥沙利铂在细胞水平上的作用机制相似。
