The PI3K/mTOR dual inhibitor NVP-BEZ235 reduces the growth of ovarian clear cell carcinoma

Oishi,Tetsuro; Itamochi,Hiroaki; Kudoh,Akiko; Nonaka,Michiko; Kato,Misaki; Nishimura,Mayumi; Oumi,Nao; Sato,Seiya; Naniwa,Jun; Sato,Shinya; Shimada,Muneaki; Kigawa,Junzo; Harada,Tasuku

doi:10.3892/or.2014.3268

The PI3K/mTOR dual inhibitor NVP-BEZ235 reduces the growth of ovarian clear cell carcinoma

Authors:
- Tetsuro Oishi
- Hiroaki Itamochi
- Akiko Kudoh
- Michiko Nonaka
- Misaki Kato
- Mayumi Nishimura
- Nao Oumi
- Seiya Sato
- Jun Naniwa
- Shinya Sato
- Muneaki Shimada
- Junzo Kigawa
- Tasuku Harada
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Affiliations: Department of Obstetrics and Gynecology, Tottori University School of Medicine, Yonago, Tottori 683-8504, Japan, Tottori University Hospital Cancer Center, Yonago, Tottori 683-8504, Japan
Published online on: June 13, 2014 https://doi.org/10.3892/or.2014.3268
Pages: 553-558

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Abstract

Patients with clear cell carcinoma of the ovary (OCCC) have poor survival due to resistance to standard chemotherapy. OCCC has frequent activating mutations of the PIK3CA gene. The present study was conducted to clarify the efficacy of the inhibition of the PI3K-AKT-mTOR pathway in OCCC. We used 8 OCCC cell lines and 5 ovarian serous adenocarcinoma (OSAC) cell lines. The mutation status of the PIK3CA and KRAS genes was examined by direct sequencing. The IC50 values of NVP-BEZ235 (BEZ235) and temsirolimus were determined by WST-8 assay. Protein expression levels of PI3K-AKT-mTOR pathway molecules were examined by western blotting. Cell cycle distribution was analyzed by flow cytometry. Annexin V staining was used for detecting apoptosis. We also investigated the effects of BEZ235 on OCCC tumor growth in a nude mouse xenograft model. Four of the 8 OCCC cell lines showed a PIK3CA mutation while none of the 5 OSAC cell lines showed a mutation. The IC50 values of BEZ235 for the OCCC cell lines were lower than these values for the OSAC cell lines. The IC50 value of temsirolimus was higher than BEZ235 in the OCCC cell lines. The PIK3CA mutation was more frequently noted in OCCC than OSAC cells, but the sensitivity of these cell lines to BEZ235 or temsirolimus was not related to the mutation status. pHER3 and pAkt proteins were expressed more frequently in OCCC compared with OSAC. However, protein expression levels were distributed widely, and were not related to the sensitivity. Treatment with BEZ235 suppressed expression of pAkt, although treatment with temsirolimus did not. OCCC cells exhibited G1 phase arrest after treatment with BEZ235 and apoptosis with a higher concentration of the agent. BEZ235 significantly inhibited tumor growth in mice bearing OVISE and TU-OC-1 cell tumors. The present study indicated that the PI3K-AKT-mTOR pathway is a potential target for OCCC, and that BEZ235 warrants investigation as a therapeutic agent.

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1	Scully RE: World Health Organization classification and nomenclature of ovarian cancer. J Natl Cancer Inst Monogr. 42:5–7. 1975.PubMed/NCBI
2	McGuire V, Jesser CA and Whittemore AS: Survival among U.S. women with invasive epithelial ovarian cancer. Gynecol Oncol. 84:399–403. 2002. View Article : Google Scholar : PubMed/NCBI
3	Kennedy AW, Biscotti CV, Hart WR and Webster KD: Ovarian clear cell adenocarcinoma. Gynecol Oncol. 32:342–349. 1989. View Article : Google Scholar
4	Sugiyama T, Kamura T, Kigawa J, et al: Clinical characteristics of clear cell carcinoma of the ovary: a distinct histologic type with poor prognosis and resistance to platinum-based chemotherapy. Cancer. 88:2584–2589. 2000. View Article : Google Scholar : PubMed/NCBI
5	Courtney KD, Corcoran RB and Engelman JA: The PI3K pathway as drug target in human cancer. J Clin Oncol. 28:1075–1083. 2010. View Article : Google Scholar : PubMed/NCBI
6	Nakayama K, Nakayama N, Kurman RJ, et al: Sequence mutations and amplification of PIK3CA and AKT2 genes in purified ovarian serous neoplasms. Cancer Biol Ther. 5:779–785. 2006. View Article : Google Scholar : PubMed/NCBI
7	Zhang L, Huang J, Yang N, et al: Integrative genomic analysis of phosphatidylinositol 3′-kinase family identifies PIK3R3 as a potential therapeutic target in epithelial ovarian cancer. Clin Cancer Res. 13:5314–5321. 2007.
8	Campbell IG, Russell SE, Choong DYH, et al: Mutation of the PIK3CA gene in ovarian and breast cancer. Clin Cancer Res. 64:7678–7681. 2004.
9	Altomare DA, Wang HQ, Skele KL, et al: AKT and mTOR phosphorylation is frequently detected in ovarian cancer and can be targeted to disrupt ovarian tumor cell growth. Oncogene. 23:5853–5857. 2004. View Article : Google Scholar : PubMed/NCBI
10	Mabuchi S, Kawase C, Altomare DA, et al: mTOR is a promising therapeutic target both in cisplatin-sensitive and cisplatin-resistant clear cell carcinoma of the ovary. Clin Cancer Res. 15:5404–5413. 2009. View Article : Google Scholar : PubMed/NCBI
11	Kuo KT, Mao TL, Jones S, et al: Frequent activating mutations of PIK3CA in ovarian clear cell carcinoma. Am J Pathol. 174:1597–1601. 2009. View Article : Google Scholar : PubMed/NCBI
12	Maira SM, Stauffer F, Brueggen J, et al: Identification and characterization of NVP-BEZ235, a new orally available dual phosphatidylinositol 3-kinase/mammalian target of rapamycin inhibitor with potent in vivo antitumor activity. Mol Cancer Ther. 7:1851–1863. 2008. View Article : Google Scholar : PubMed/NCBI
13	Brachmann S, Hofmann I, Schnell C, et al: Specific apoptosis induction by the dual PI3K/mTOR inhibitor NVP-BEZ235 in HER2 amplified and PIK3CA mutant breast cancer cells. Proc Natl Acad Sci. 106:22299–22304. 2009. View Article : Google Scholar : PubMed/NCBI
14	Cho DC, Cohen MB, Panka DJ, et al: The efficacy of the novel dual PI3-kinase/mTOR inhibitor NVP-BEZ235 compared with rapamycin in renal cell carcinoma. Clin Cancer Res. 16:3628–3638. 2010. View Article : Google Scholar : PubMed/NCBI
15	Itamochi H, Kato M, Nishimura M, et al: Establishment and characterization of a novel ovarian serous adenocarcinoma cell line, TU-OS-4, that overexpresses EGFR and HER2. Hum Cell. 25:111–115. 2012. View Article : Google Scholar : PubMed/NCBI
16	Itamochi H, Kato M, Nishimura M, et al: Establishment and characterization of a novel ovarian clear cell carcinoma cell line, TU-OC-1, with a mutation in the PIK3CA gene. Hum Cell. 26:121–127. 2013. View Article : Google Scholar : PubMed/NCBI
17	Itamochi H, Oishi T, Shimada M, et al: Inhibiting the mTOR pathway synergistically enhances cytotoxicity in ovarian cancer cells induced by etoposide through upregulation of c-Jun. Clin Cancer Res. 17:4742–4750. 2011. View Article : Google Scholar : PubMed/NCBI
18	McIntyre AJ, Summersgill BM, Spendlove HE, et al: Activating mutations and/or expression levels of tyrosine kinase receptors GRB7, RAS, and BRAF in testicular germ cell tumors. Neoplasia. 7:1047–1052. 2005. View Article : Google Scholar : PubMed/NCBI
19	Li VSW, Wong CW, Chan TL, et al: Mutations of PIK3CA in gastric adenocarcinoma. BMC Cancer. 5:292005. View Article : Google Scholar
20	Rauh-Hein AJ, Winograd D, Growdon WB, et al: Prognostic determinants in patients with uterine and ovarian clear carcinoma. Gynecol Oncol. 125:376–380. 2012. View Article : Google Scholar : PubMed/NCBI
21	Pectasides D, Fountzilas G, Aravantinos G, et al: Advanced stage clear-cell epithelial ovarian cancer: the Hellenic Cooperative Oncology Group experience. Gynecol Oncol. 102:285–291. 2006. View Article : Google Scholar : PubMed/NCBI
22	Miyazawa M, Yasuda M, Fujita M, et al: Therapeutic strategy targeting the mTOR-HIF-1α-VEGF pathway in ovarian clear cell adenocarcinoma. Pathol Int. 59:19–27. 2009.
23	Itamochi H and Kigawa J: Clinical trials and future potential of targeted therapy for ovarian cancer. Int J Clin Oncol. 17:430–440. 2012. View Article : Google Scholar : PubMed/NCBI
24	Efeyan A and Sabatini DM: mTOR and cancer: many loops in one pathway. Curr Opin Cell Biol. 22:169–176. 2010. View Article : Google Scholar : PubMed/NCBI
25	Harrington LS, Findlay GM, Gray A, et al: The TSC1-2 tumor suppressor controls insulin-PI3K signaling via regulation of IRS proteins. J Cell Biol. 166:213–223. 2004. View Article : Google Scholar : PubMed/NCBI
26	Hartley D and Cooper GM: Role of mTOR in the degradation of IRS-1: regulation of PP2A activity. J Cell Biochem. 85:304–314. 2002. View Article : Google Scholar : PubMed/NCBI
27	Shoji K, Oda K, Kashiyama T, et al: Genotype-dependent efficacy of a dual PI3K/mTOR inhibitor, NVP-BEZ235, and an mTOR inhibitor, RAD001, in endometrial carcinomas. PLoS One. 7:e374312012. View Article : Google Scholar : PubMed/NCBI
28	Montero JC, Chen X, Ocaria A, et al: Predominance of mTORC1 over mTORC2 in the regulation of proliferation of ovarian cancer cells: therapeutic implications. Mol Cancer Ther. 11:1342–1352. 2012. View Article : Google Scholar : PubMed/NCBI
29	Santiskulvong C, Konecny GE, Fekete M, et al: Dual targeting of phosphoinositide 3-kinase and mammalian target of rapamycin using NVP-BEZ235 as a novel therapeutic approach in human ovarian carcinoma. Clin Cancer Res. 17:2373–2384. 2011. View Article : Google Scholar : PubMed/NCBI
30	Rahman M, Nakayama K, Rahman MT, et al: Clinicopathologic and biological analysis of PIK3CA mutation in ovarian clear cell carcinoma. Hum Pathol. 43:2197–2206. 2012.PubMed/NCBI