Topotecan synergizes with CHEK1 (CHK1) inhibitor to induce apoptosis in ovarian cancer cells

BMC Cancer

Marianne K Kim¹  Jana James¹  Christina M Annunziata¹ 

[1] Women’s Malignancies Branch, Center for Cancer Research, National Cancer Institute, NIH, Bethesda 20892, MD, USA

关键词: Ovarian cancer;    CHEK1 (CHK1);    Topotecan;   

Others  :  1161234
DOI  :  10.1186/s12885-015-1231-z

 received in 2014-09-19, accepted in 2015-03-19,  发布年份 2015

【 摘 要 】

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.

【 授权许可】

   
2015 Kim et al.; licensee BioMed Central.

【 图 表 】

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【 参考文献 】

• [1]McCluggage WG: Morphological subtypes of ovarian carcinoma: a review with emphasis on new developments and pathogenesis. Pathol 2011, 43(5):420-32.
• [2]Cancer Genome Atlas Research Network: Integrated genomic analyses of ovarian carcinoma Nature 2011, 474(7353):609-15.
• [3]Dasari S, Bernard Tchounwou P: Cisplatin in cancer therapy: molecular mechanisms of action. Eur J Pharmacol 2014, 740C:364-78.
• [4]Slichenmyer WJ, Rowinsky EK, Donehower RC, Kaufmann SH: The current status of camptothecin analogues as antitumor agents. J Natl Cancer Inst 1993, 85(4):271-91.
• [5]Lorusso D, Pietragalla A, Mainenti S, Masciullo V, Di Vagno G, Scambia G: Review role of topotecan in gynaecological cancers: current indications and perspectives. Crit Rev Oncol Hematol 2010, 74(3):163-74.
• [6]Kim MK, Min DJ, Wright G, Goldlust I, Annunziata CM: Loss of compensatory pro-survival and anti-apoptotic modulator, IKKepsilon, sensitizes ovarian cancer cells to CHEK1 loss through an increased level of p21. Oncotarget 2014, 5(24):12788-802.
• [7]Goto H, Izawa I, Li P, Inagaki M: Novel regulation of checkpoint kinase 1: Is checkpoint kinase 1 a good candidate for anti-cancer therapy? Cancer Sci 2012, 103(7):1195-200.
• [8]Rawlinson R, Massey AJ: GammaH2AX and Chk1 phosphorylation as predictive pharmacodynamic biomarkers of Chk1 inhibitor-chemotherapy combination treatments. BMC Cancer 2014, 14:483. BioMed Central Full Text
• [9]Parsels LA, Qian Y, Tanska DM, Gross M, Zhao L, Hassan MC, et al.: Assessment of chk1 phosphorylation as a pharmacodynamic biomarker of chk1 inhibition. Clin Cancer Res 2011, 17(11):3706-15.
• [10]Dent P, Tang Y, Yacoub A, Dai Y, Fisher PB, Grant S: CHK1 inhibitors in combination chemotherapy: thinking beyond the cell cycle. Mol Interv 2011, 11(2):133-40.
• [11]Tao ZF, Lin NH: Chk1 inhibitors for novel cancer treatment. Anticancer Agents Med Chem 2006, 6(4):377-88.
• [12]Domcke S, Sinha R, Levine DA, Sander C, Schultz N: Evaluating cell lines as tumour models by comparison of genomic profiles. Nat Commun 2013, 4:2126.
• [13]Langdon SP, Lawrie SS, Hay FG, Hawkes MM, McDonald A, Hayward IP, et al.: Characterization and properties of nine human ovarian adenocarcinoma cell lines. Cancer Res 1988, 48(21):6166-72.
• [14]Barretina J, Caponigro G, Stransky N, Venkatesan K, Margolin AA, Kim S, et al.: The Cancer Cell Line Encyclopedia enables predictive modelling of anticancer drug sensitivity. Nature 2012, 483(7391):603-7.
• [15]Blasina A, Hallin J, Chen E, Arango ME, Kraynov E, Register J, et al.: Breaching the DNA damage checkpoint via PF-00477736, a novel small-molecule inhibitor of checkpoint kinase 1. Mol Cancer Ther 2008, 7(8):2394-404.
• [16]Aris SM, Pommier Y: Potentiation of the novel topoisomerase I inhibitor indenoisoquinoline LMP-400 by the cell checkpoint and Chk1-Chk2 inhibitor AZD7762. Cancer Res 2012, 72(4):979-89.
• [17]Huntoon CJ, Flatten KS, Wahner Hendrickson AE, Huehls AM, Sutor SL, Kaufmann SH, et al.: ATR inhibition broadly sensitizes ovarian cancer cells to chemotherapy independent of BRCA status. Cancer Res 2013, 73(12):3683-91.
• [18]Xiao Y, Ramiscal J, Kowanetz K, Del Nagro C, Malek S, Evangelista M, et al.: Identification of preferred chemotherapeutics for combining with a CHK1 inhibitor. Mol Cancer Ther 2013, 12(11):2285-95.
• [19]Walton MI, Eve PD, Hayes A, Valenti MR, De Haven Brandon AK, Box G, et al.: CCT244747 is a novel potent and selective CHK1 inhibitor with oral efficacy alone and in combination with genotoxic anticancer drugs. Clin Cancer Res 2012, 18(20):5650-61.
• [20]Daud AI, Ashworth MT, Strosberg J, Goldman JW, Mendelson D, Springett G, et al.: Phase I dose-escalation trial of checkpoint kinase 1 inhibitor MK-8776 as Monotherapy and in combination with gemcitabine in patients with advanced solid tumors. J Clin Oncol 2015, 33(9):1060-6.
• [21]Pujade-Lauraine E, Hilpert F, Weber B, Reuss A, Poveda A, Kristensen G, et al.: Bevacizumab combined with chemotherapy for platinum-resistant recurrent ovarian cancer: the AURELIA open-label randomized phase III trial. J Clin Oncol 2014, 32(13):1302-8.
• [22]Vergote I, Schilder RJ, Pippitt CH Jr, Wong S, Gordon AN, Scudder S, et al.: A phase 1b study of trebananib in combination with pegylated liposomal doxorubicin or topotecan in women with recurrent platinum-resistant or partially platinum-sensitive ovarian cancer. Gynecol Oncol 2014, 135(1):25-33.
• [23]Lee JM, Hays JL, Annunziata CM, Noonan AM, Minasian L, Zujewski JA, et al.: Phase I/Ib study of olaparib and carboplatin in BRCA1 or BRCA2 mutation-associated breast or ovarian cancer with biomarker analyses. J Natl Cancer Inst 2014, 106(6):dju089.
• [24]Bryant C, Rawlinson R, Massey AJ: Chk1 inhibition as a novel therapeutic strategy for treating triple-negative breast and ovarian cancers. BMC Cancer 2014, 14:570. BioMed Central Full Text