Mammalian target of rapamycin inhibitor RAD001 sensitizes endometrial cancer cells to paclitaxel-induced apoptosis via the induction of autophagy

Wang,Huan; Li,Dandan; Li,Xiaomao; Ou,Xueling; Liu,Suiling; Zhang,Yu; Ding,Jie; Xie,Bo

doi:10.3892/ol.2016.5338

Mammalian target of rapamycin inhibitor RAD001 sensitizes endometrial cancer cells to paclitaxel-induced apoptosis via the induction of autophagy

Authors:
- Huan Wang
- Dandan Li
- Xiaomao Li
- Xueling Ou
- Suiling Liu
- Yu Zhang
- Jie Ding
- Bo Xie
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Affiliations: Department of Gynecology, Third Affiliated Hospital of Sun Yat‑sen University, Guangzhou, Guangdong 510630, P.R. China, Sun Yat‑sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou, Guangdong 510060, P.R. China, Department of Forensic Medicine, Zhongshan School of Medicine, Sun Yat‑sen University, Guangzhou, Guangdong 510089, P.R. China, Department of Pharmacology, Zhongshan School of Medicine, Sun Yat‑sen University, Guangzhou, Guangdong 510089, P.R. China
Published online on: November 2, 2016 https://doi.org/10.3892/ol.2016.5338
Pages: 5029-5035
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to investigate the effects of the mammalian target of rapamycin (mTOR) inhibitor, RAD001, on the growth of human endometrial cancer cells. The effects of RAD001 on human endometrial cancer Ishikawa and HEC-1A cell proliferation were determined by MTT assay. Green fluorescent protein microtubule-associated protein 1 light chain 3α (GFP-LC3) protein aggregates were observed under a confocal microscope, and Ishikawa and HEC‑1A cell apoptosis was detected using flow cytometry. The expression levels of LC3‑I, LC3-II and mTOR proteins were detected by western blot analysis. The results showed that RAD001 effectively inhibited human endometrial cancer Ishikawa and HEC‑1A cell proliferation via downregulation of AKT/mTOR phosphorylation. Moreover, RAD001 induced autophagic cell death and a higher sensitivity to paclitaxel-induced apoptosis. These results indicate that RAD001 could have therapeutic potential in human endometrial cancer with hyperactivated AKT/mTOR signaling.

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1	Jemal A, Siegel R, Xu J and Ward E: Cancer statistics, 2010. CA Cancer J Clin. 60:277–300. 2010. View Article : Google Scholar : PubMed/NCBI
2	Siegel R, Naishadham D and Jemal A: Cancer statistics, 2012. CA Cancer J Clin. 62:10–29. 2012. View Article : Google Scholar : PubMed/NCBI
3	Sankaranarayanan R and Ferlay J: Worldwide burden of gynaecological cancer: The size of the problem. Best Pract Res Clin Obstet Gynaecol. 20:207–225. 2006. View Article : Google Scholar : PubMed/NCBI
4	Boya P, González-Polo RA, Casares N, Perfettini JL, Dessen P, Larochette N, Métivier D, Meley D, Souquere S, Yoshimori T, et al: Inhibition of macroautophagy triggers apoptosis. Mol Cell Biol. 25:1025–1040. 2005. View Article : Google Scholar : PubMed/NCBI
5	Qi XF, Kim DH, Lee KJ, Kim CS, Song SB, Cai DQ and Kim SK: Autophagy contributes to apoptosis in A20 and EL4 lymphoma cells treated with fluvastatin. Cancer Cell Int. 13:1112013. View Article : Google Scholar : PubMed/NCBI
6	Kondo Y, Kanzawa T, Sawaya R and Kondo S: The role of autophagy in cancer development and response to therapy. Nat Rev Cancer. 5:726–734. 2005. View Article : Google Scholar : PubMed/NCBI
7	Mathew R, Karantza-Wadsworth V and White E: Role of autophagy in cancer. Nat Rev Cancer. 7:961–967. 2007. View Article : Google Scholar : PubMed/NCBI
8	Jung CH, Ro SH, Cao J, Otto NM and Kim DH: mTOR regulation of autophagy. FEBS Lett. 584:1287–1295. 2010. View Article : Google Scholar : PubMed/NCBI
9	Ganley IG, Lam du H, Wang J, Ding X, Chen S and Jiang X: ULK1 ATG13. FIP200 complex mediates mTOR signaling and is essential for autophagy. J Biol Chem. 284:12297–12305. 2009. View Article : Google Scholar : PubMed/NCBI
10	Guertin DA and Sabatini DM: Defining the role of mTOR in cancer. Cancer Cell. 12:9–22. 2007. View Article : Google Scholar : PubMed/NCBI
11	Hippert MM, O'Toole PS and Thorburn A: Autophagy in cancer: Good, bad, or both? Cancer Res. 66:9349–9351. 2006. View Article : Google Scholar : PubMed/NCBI
12	Lévy A, Sauvin LA, Massard C and Soria JC: Everolimus (RAD001) and solid tumours: A 2008 summary. Bull Cancer. 95:1205–1211. 2008.(In French). PubMed/NCBI
13	Agarwala SS and Case S: Everolimus (RAD001) in the treatment of advanced renal cell carcinoma: A review. Oncologist. 15:236–245. 2010. View Article : Google Scholar : PubMed/NCBI
14	Beuvink I, Boulay A, Fumagalli S, Zilbermann F, Ruetz S, O'Reilly T, Natt F, Hall J, Lane HA and Thomas G: The mTOR inhibitor RAD001 sensitizes tumor cells to DNA-damaged induced apoptosis through inhibition of p21 translation. Cell. 120:747–759. 2005. View Article : Google Scholar : PubMed/NCBI
15	Weigelt B and Downward J: Genomic determinants of PI3K pathway inhibitor response in cancer. Front Oncol. 2:1092012. View Article : Google Scholar : PubMed/NCBI
16	Dengler J, von Bubnoff N, Decker T, Peschel C and Duyster J: Combination of imatinib with rapamycin or RAD001 acts synergistically only in Bcr-Abl-positive cells with moderate resistance to imatinib. Leukemia. 19:1835–1838. 2005. View Article : Google Scholar : PubMed/NCBI
17	Cao C, Subhawong T, Albert JM, Kim KW, Geng L, Sekhar KR, Gi YJ and Lu B: Inhibition of mammalian target of rapamycin or apoptotic pathway induces autophagy and radiosensitizes PTEN null prostate cancer cells. Cancer Res. 66:10040–10047. 2006. View Article : Google Scholar : PubMed/NCBI
18	Shinohara ET, Cao C, Niermann K, Mu Y, Zeng F, Hallahan DE and Lu B: Enhanced radiation damage of tumor vasculature by mTOR inhibitors. Oncogene. 24:5414–5422. 2005. View Article : Google Scholar : PubMed/NCBI
19	Haritunians T, Mori A, O'Kelly J, Luong QT, Giles FJ and Koeffler HP: Antiproliferative activity of RAD001 (everolimus) as a single agent and combined with other agents in mantle cell lymphoma. Leukemia. 21:333–339. 2007. View Article : Google Scholar : PubMed/NCBI
20	Panner A, Parsa AT and Pieper RO: Use of APO2L/TRAIL with mTOR inhibitors in the treatment of glioblastoma multiforme. Expert Rev Anticancer Ther. 6:1313–1322. 2006. View Article : Google Scholar : PubMed/NCBI
21	Mayer IA, Burris H, Bendel J, Means-Powell J, Arteaga C, Shyr Y and Pietenpol J: A phase Ib trial of RAD001, an mTOR inhibitor, with weekly cisplatin and paclitaxel in patients with HER2-negative metastatic breast cancer. Abstract 3093. 32nd Annual San Antonio Breast Cancer Symposium. San Antonio, TX. December 9–13, 2009;
22	Mason RL, Gunst RF and Hess JJ: Statistical Design and Analysis of ExperimentsWith Applications to Engineering and Science. 2nd. John Wiley and Sons Inc.; Hoboken, NJ: 2003
23	Tanida I, Ueno T and Kominami E: LC3 and autophagy. Methods Mol Biol. 445:77–88. 2008. View Article : Google Scholar : PubMed/NCBI
24	Sever R and Brugge JS: Signal transduction in cancer. Cold Spring Harb Perspect Med. 5:a0060982015. View Article : Google Scholar : PubMed/NCBI
25	Kolch W, Halasz M, Granovskaya M and Kholodenko BN: The dynamic control of signal transduction networks in cancer cells. Nat Rev Cancer. 15:515–527. 2015. View Article : Google Scholar : PubMed/NCBI
26	Ranjan A, Fofaria NM, Kim SH and Srivastava SK: Modulation of signal transduction pathways by natural compounds in cancer. Chin J Nat Med. 13:730–742. 2015.PubMed/NCBI
27	Zhou HY and Huang SL: Current development of the second generation of mTOR inhibitors as anticancer agents. Chin J Cancer. 31:8–18. 2012.PubMed/NCBI
28	Tan HK, Moad AI and Tan ML: The mTOR signalling pathway in cancer and the potential mTOR inhibitory activities of natural phytochemicals. Asian Pac J Cancer Prev. 15:6463–6475. 2014. View Article : Google Scholar : PubMed/NCBI
29	Ozols RF: Paclitaxel (Taxol)/carboplatin combination chemotherapy in the treatment of advanced ovarian cancer. Semin Oncol 27 (Suppl 7). 3–7. 2000.
30	Okamoto I, Moriyama E, Fujii S, Kishi H, Nomura M, Goto E, Kiyofuji C, Imamura F, Mori T and Matsumoto M: Phase II study of carboplatin-paclitaxel combination chemotherapy in elderly patients with advanced non-small cell lung cancer. Jpn J Clin Oncol. 35:188–194. 2005. View Article : Google Scholar : PubMed/NCBI
31	Pentheroudakis G, Razis E, Athanassiadis A, Pavlidis N and Fountzilas G: Paclitaxel-carboplatin combination chemotherapy in advanced breast cancer: Accumulating evidence for synergy, efficacy, and safety. Med Oncol. 23:147–160. 2006. View Article : Google Scholar : PubMed/NCBI
32	Yardley DA, Brufsky A, Coleman RE, Conte PF, Cortes J, Glück S, Nabholtz JM, O'Shaughnessy J, Beck RM, Ko A, et al: Phase II/III weekly nab-paclitaxel plus gemcitabine or carboplatin versus gemcitabine/carboplatin as first-line treatment of patients with metastatic triple-negative breast cancer (the tnAcity study): Study protocol for a randomized controlled trial. Trials. 16:5752015. View Article : Google Scholar : PubMed/NCBI
33	Yamazaki S, Sekine I and Saijo N: Paclitaxel (taxol): A review of its antitumor activity and toxicity in clinical studies. Gan To Kagaku Ryoho. 25:605–615. 1998.(In Japanese). PubMed/NCBI
34	Liu H, Chen X, Sun J, Gao P, Song Y, Zhang N, Lu X, Xu H and Wang Z: The efficacy and toxicity of paclitaxel plus S-1 compared with paclitaxel plus 5-FU for advanced gastric cancer: A PRISMA systematic review and meta-analysis of randomized controlled trials. Medicine (Baltimore). 93:e1642014. View Article : Google Scholar : PubMed/NCBI
35	Dufour M, Dormond-Meuwly A, Demartines N and Dormond O: Targeting the Mammalian Target of Rapamycin (mTOR) in Cancer Therapy: Lessons from Past and Future Perspectives. Cancers (Basel). 3:2478–2500. 2011. View Article : Google Scholar : PubMed/NCBI
36	Mizushima N: Autophagy: Process and function. Genes Dev. 21:2861–2873. 2007. View Article : Google Scholar : PubMed/NCBI
37	Berry DL and Baehrecke EH: Autophagy functions in programmed cell death. Autophagy. 4:359–360. 2008. View Article : Google Scholar : PubMed/NCBI
38	Alonso S, Pethe K, Russell DG and Purdy GE: Lysosomal killing of Mycobacterium mediated by ubiquitin-derived peptides is enhanced by autophagy. Proc Natl Acad Sci USA. 104:6031–6036. 2007. View Article : Google Scholar : PubMed/NCBI
39	Lum JJ, Bauer DE, Kong M, Harris MH, Li C, Lindsten T and Thompson CB: Growth factor regulation of autophagy and cell survival in the absence of apoptosis. Cell. 120:237–248. 2005. View Article : Google Scholar : PubMed/NCBI
40	Park HJ, Lee SJ, Kim SH, Han J, Bae J, Kim SJ, Park CG and Chun T: IL-10 inhibits the starvation induced autophagy in macrophages via class I phosphatidylinositol 3-kinase (PI3K) pathway. Mol Immunol. 48:720–727. 2011. View Article : Google Scholar : PubMed/NCBI
41	Pattingre S, Tassa A, Qu X, Garuti R, Liang XH, Mizushima N, Packer M, Schneider MD and Levine B: Bcl-2 antiapoptotic proteins inhibit Beclin 1-dependent autophagy. Cell. 122:927–939. 2005. View Article : Google Scholar : PubMed/NCBI