Pharmacologic inhibition of ATR and ATM offers clinically important distinctions to enhancing platinum or radiation response in ovarian, endometrial, and cervical cancer cells
Section snippets
Background
Gynecologic (GYN) cancers, specifically cancers of the uterine cervix, uterine corpus, and ovary, represent a significant worldwide health problem with 1,085,948 new diagnoses and 493,713 deaths observed annually (globocan.iarc.fr). Primary cytoreductive surgery with platinum- and paclitaxel-based chemotherapy, either in the adjuvant or neoadjuvant setting, are the standards of care for ovarian cancer [1]. Surgical staging plays a central role in endometrial cancer management, with
Cell culture
A2780 and A2780-CP20 (Sigma, St. Louis, MO) and maintained in RPMI medium supplemented with 10% fetal bovine serum (FBS, ATCC, Manassas, VA). A2780-CP20 cells were treated with 1 μM cisplatin (Sigma) every other passage to maintain resistance to cisplatin. OVCAR3 cells maintained in RPMI medium supplemented with 20% FBS and 0.01 mg/mL insulin (Sigma) or DMEM:F12 with 10% FBS and 0.01 mg/mL insulin. KLE cells were maintained in DMEM:F12 medium supplemented with 10% FBS. HELA, SIHA, and HEC1B cells
Analysis of DDR protein expression across representative cell line models of gynecologic cancer
Protein abundances of ATR, ATM, Chk1, Chk2, and p53 were analyzed by immunoblot in seven cell line models of GYN carcinoma, including three models of ovarian [A2780, A2780-CP20 (a syngeneic model of cisplatin resistance derived from A2780), and OVCAR3], two models of cervical (HELA and SIHA) and two models of endometrial (KLE and HEC1B) carcinoma (Supplementary Fig. 1). Total ATR levels were largely invariant in these cells, with the exception of OVCAR3 where a greater level of ATR abundance
Discussion
Here we show that pharmacological inhibition of ATR, but not ATM, sensitizes ovarian, endometrial and cervical cancer cells to cisplatin and carboplatin, and that the combination of either platinum agent with the ATRi was synergistic. We present data utilizing a syngeneic model of platinum-resistant ovarian cancer, A2780 (platinum-sensitive) versus A2780-CP20 (platinum-resistant), showing that inhibition of ATR produces marked enhancement in response when combined with cisplatin or carboplatin.
Funding
This study was funded by an award from the United States Army Medical Research and Materiel Command (W81XWH-11-2-0131, all authors but CJB) and CA148644 (CJB) from the National Cancer Institute (CJB).
Competing interests
The opinions or assertions contained herein are the private views of the authors and are not to be construed as official or as reflecting the views of the Department of the Army, Department of the Air Force, Department of the Navy, or the Department of Defense. The authors have no financial disclosures.
Conflict of interest statement
The authors declare that there are no conflicts of interest.
Acknowledgments
The authors thank Darcy Franicola for her assistance in generating the IR clonogenic plots.
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These authors contributed equally to this work.