Dihydrocelastrol exerts potent antitumor activity in mantle cell lymphoma cells via dual inhibition of mTORC1 and mTORC2

Xie,Yongsheng; Li,Bo; Bu,Wenxuan; Gao,Lu; Zhang,Yong; Lan,Xiucai; Hou,Jun; Xu,Zhijian; Chang,Shuaikang; Yu,Dandan; Xie,Bingqian; Wang,Yingcong; Wang,Houcai; Zhang,Yiwen; Wu,Xiaosong; Zhu,Weiliang; Shi,Jumei

doi:10.3892/ijo.2018.4438

Dihydrocelastrol exerts potent antitumor activity in mantle cell lymphoma cells via dual inhibition of mTORC1 and mTORC2

Authors:
- Yongsheng Xie
- Bo Li
- Wenxuan Bu
- Lu Gao
- Yong Zhang
- Xiucai Lan
- Jun Hou
- Zhijian Xu
- Shuaikang Chang
- Dandan Yu
- Bingqian Xie
- Yingcong Wang
- Houcai Wang
- Yiwen Zhang
- Xiaosong Wu
- Weiliang Zhu
- Jumei Shi
View Affiliations

Affiliations: Department of Hematology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai 200072, P.R. China, CAS Key Laboratory of Receptor Research, Drug Discovery and Design Center, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, P.R. China
Published online on: June 12, 2018 https://doi.org/10.3892/ijo.2018.4438
Pages: 823-834

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Mantle cell lymphoma (MCL) is a distinct and highly aggressive subtype of B-cell non-Hodgkin lymphoma. Dihydrocelastrol (DHCE) is a dihydro-analog of celastrol, which is isolated from the traditional Chinese medicinal plant Tripterygium wilfordii. The present study aimed to investigate the effects of DHCE treatment on MCL cells, and to determine the mechanism underlying its potent antitumor activity in vitro and in vivo using the Cell Counting kit-8 assay, clonogenic assay, apoptosis assay, cell cycle analysis, immunofluorescence staining, western blotting and tumor xenograft models. The results demonstrated that DHCE treatment exerted minimal cytotoxic effects on normal cells, but markedly suppressed MCL cell proliferation by inducing G0/G1 phase cell cycle arrest, and inhibited MCL cell viability by stimulating apoptosis via extrinsic and intrinsic pathways. In addition, the results revealed that DHCE suppressed cell growth and proliferation by inhibiting mammalian target of rapamycin complex (mTORC)1-mediated phosphorylation of ribosomal protein S6 kinase and eukaryotic initiation factor 4E binding protein. Simultaneously, DHCE induced apoptosis and inhibited cell survival by suppressing mTORC2-mediated phosphorylation of protein kinase B and nuclear factor-κB activity. In addition to in vitro findings, DHCE treatment reduced the MCL tumor burden in a xenograft mouse model, without indications of toxicity. Furthermore, combined treatment with DHCE and bortezomib, a proteasome inhibitor, induced a synergistic cytotoxic effect on MCL cells. These findings indicated that DHCE may have the potential to serve as a novel therapeutic agent for the treatment of MCL through dually inhibiting mTORC1 and mTORC2.

View Figures

View References

1	Spurgeon SE, Till BG, Martin P, Goy AH, Dreyling MP, Gopal AK, LeBlanc M, Leonard JP, Friedberg JW, Baizer L, et al: Recommendations for clinical trial development in mantle cell lymphoma. J Natl Cancer Inst. 109:1092016.
2	Zhou Y, Wang H, Fang W, Romaguer JE, Zhang Y, Delasalle KB, Kwak L, Yi Q, Du XL and Wang M: Incidence trends of mantle cell lymphoma in the United States between 1992 and 2004. Cancer. 113:791–798. 2008. View Article : Google Scholar : PubMed/NCBI
3	Bosch F, Jares P, Campo E, Lopez-Guillermo A, Piris MA, Villamor N, Tassies D, Jaffe ES, Montserrat E, Rozman C, et al: PRAD-1/cyclin D1 gene overexpression in chronic lymphoproliferative disorders: A highly specific marker of mantle cell lymphoma. Blood. 84:2726–2732. 1994.PubMed/NCBI
4	Cheah CY, Seymour JF and Wang ML: Mantle cell lymphoma. J Clin Oncol. 34:1256–1269. 2016. View Article : Google Scholar : PubMed/NCBI
5	Hamad N, Armytage T, McIlroy K, Singh N and Ward C: Primary cutaneous mantle-cell lymphoma: A case report and literature review. J Clin Oncol. 33:e104–e108. 2015. View Article : Google Scholar
6	Campo E and Rule S: Mantle cell lymphoma: Evolving management strategies. Blood. 125:48–55. 2015. View Article : Google Scholar
7	Lee JS, Vo TT and Fruman DA: Targeting mTOR for the treatment of B cell malignancies. Br J Clin Pharmacol. 82:1213–1228. 2016. View Article : Google Scholar : PubMed/NCBI
8	Strimpakos AS, Karapanagiotou EM, Saif MW and Syrigos KN: The role of mTOR in the management of solid tumors: An overview. Cancer Treat Rev. 35:148–159. 2009. View Article : Google Scholar
9	Baldo P, Cecco S, Giacomin E, Lazzarini R, Ros B and Marastoni S: mTOR pathway and mTOR inhibitors as agents for cancer therapy. Curr Cancer Drug Targets. 8:647–665. 2008. View Article : Google Scholar : PubMed/NCBI
10	Inoki K, Corradetti MN and Guan KL: Dysregulation of the TSC-mTOR pathway in human disease. Nat Genet. 37:19–24. 2005. View Article : Google Scholar
11	Lee DF, Kuo HP, Chen CT, Hsu JM, Chou CK, Wei Y, Sun HL, Li LY, Ping B, Huang WC, et al: IKK beta suppression of TSC1 links inflammation and tumor angiogenesis via the mTOR pathway. Cell. 130:440–455. 2007. View Article : Google Scholar : PubMed/NCBI
12	Eyre TA, Collins GP, Goldstone AH and Cwynarski K: Time now to TORC the TORC? New developments in mTOR pathway inhibition in lymphoid malignancies. Br J Haematol. 166:336–351. 2014. View Article : Google Scholar : PubMed/NCBI
13	Peponi E, Drakos E, Reyes G, Leventaki V, Rassidakis GZ and Medeiros LJ: Activation of mammalian target of rapamycin signaling promotes cell cycle progression and protects cells from apoptosis in mantle cell lymphoma. Am J Pathol. 169:2171–2180. 2006. View Article : Google Scholar : PubMed/NCBI
14	Saxton RA and Sabatini DM: mTOR signaling in growth, metabolism, and disease. Cell. 168:960–976. 2017. View Article : Google Scholar : PubMed/NCBI
15	Dancey J: mTOR signaling and drug development in cancer. Nat Rev Clin Oncol. 7:209–219. 2010. View Article : Google Scholar : PubMed/NCBI
16	Sarbassov DD, Guertin DA, Ali SM and Sabatini DM: Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex. Science. 307:1098–1101. 2005. View Article : Google Scholar : PubMed/NCBI
17	Gupta M, Hendrickson AE, Yun SS, Han JJ, Schneider PA, Koh BD, Stenson MJ, Wellik LE, Shing JC, Peterson KL, et al: Dual mTORC1/mTORC2 inhibition diminishes Akt activation and induces Puma-dependent apoptosis in lymphoid malignancies. Blood. 119:476–487. 2012. View Article : Google Scholar :
18	Witzig TE, Geyer SM, Ghobrial I, Inwards DJ, Fonseca R, Kurtin P, Ansell SM, Luyun R, Flynn PJ, Morton RF, et al: Phase II trial of single-agent temsirolimus (CCI-779) for relapsed mantle cell lymphoma. J Clin Oncol. 23:5347–5356. 2005. View Article : Google Scholar : PubMed/NCBI
19	Ansell SM, Inwards DJ, Rowland KM Jr, Flynn PJ, Morton RF, Moore DF Jr, Kaufmann SH, Ghobrial I, Kurtin PJ, Maurer M, et al: Low-dose, single-agent temsirolimus for relapsed mantle cell lymphoma: A phase 2 trial in the North Central Cancer Treatment Group. Cancer. 113:508–514. 2008. View Article : Google Scholar : PubMed/NCBI
20	Liu J, Lee J, Salazar Hernandez MA, Mazitschek R and Ozcan U: Treatment of obesity with celastrol. Cell. 161:999–1011. 2015. View Article : Google Scholar : PubMed/NCBI
21	Kannaiyan R, Shanmugam MK and Sethi G: Molecular targets of celastrol derived from Thunder of God Vine: Potential role in the treatment of inflammatory disorders and cancer. Cancer Lett. 303:9–20. 2011. View Article : Google Scholar
22	Sethi G, Ahn KS, Pandey MK and Aggarwal BB: Celastrol, a novel triterpene, potentiates TNF-induced apoptosis and suppresses invasion of tumor cells by inhibiting NF-kappaB-regulated gene products and TAK1-mediated NF-kappaB activation. Blood. 109:2727–2735. 2007.
23	Jang SY, Jang SW and Ko J: Celastrol inhibits the growth of estrogen positive human breast cancer cells through modulation of estrogen receptor α. Cancer Lett. 300:57–65. 2011. View Article : Google Scholar
24	Fribley AM, Miller JR, Brownell AL, Garshott DM, Zeng Q, Reist TE, Narula N, Cai P, Xi Y, Callaghan MU, et al: Celastrol induces unfolded protein response-dependent cell death in head and neck cancer. Exp Cell Res. 330:412–422. 2015. View Article : Google Scholar
25	Klaić L, Trippier PC, Mishra RK, Morimoto RI and Silverman RB: Remarkable stereospecific conjugate additions to the Hsp90 inhibitor celastrol. J Am Chem Soc. 133:19634–19637. 2011. View Article : Google Scholar
26	Hu L, Wu H, Li B, Song D, Yang G, Chen G, Xie B, Xu Z, Zhang Y, Yu D, et al: Dihydrocelastrol inhibits multiple myeloma cell proliferation and promotes apoptosis through ERK1/2 and IL-6/STAT3 pathways in vitro and in vivo. Acta Biochim Biophys Sin (Shanghai). 49:420–427. 2017. View Article : Google Scholar
27	Ashton JC: Drug combination studies and their synergy quantification using the Chou-Talalay method - letter. Cancer Res. 75:24002015. View Article : Google Scholar
28	Rudelius M, Rosenfeldt MT, Leich E, Rauert-Wunderlich H, Solimando AG, Beilhack A, Ott G and Rosenwald A: Inhibition of focal adhesion kinase overcomes resistance of mantle cell lymphoma to ibrutinib in the bone marrow microenvironment. Haematologica. 103:116–125. 2018. View Article : Google Scholar :
29	Zhou J, Tiemann K, Chomchan P, Alluin J, Swiderski P, Burnett J, Zhang X, Forman S, Chen R and Rossi J: Dual functional BAFF receptor aptamers inhibit ligand-induced proliferation and deliver siRNAs to NHL cells. Nucleic Acids Res. 41:4266–4283. 2013. View Article : Google Scholar : PubMed/NCBI
30	Sherr CJ and Roberts JM: CDK inhibitors: Positive and negative regulators of G1-phase progression. Genes Dev. 13:1501–1512. 1999. View Article : Google Scholar : PubMed/NCBI
31	Hassan M, Watari H, AbuAlmaaty A, Ohba Y and Sakuragi N: Apoptosis and molecular targeting therapy in cancer. BioMed Res Int. 2014:1508452014. View Article : Google Scholar : PubMed/NCBI
32	Taylor RC, Cullen SP and Martin SJ: Apoptosis: Controlled demolition at the cellular level. Nat Rev Mol Cell Biol. 9:231–241. 2008. View Article : Google Scholar
33	Davids MS: Targeting BCL-2 in B-cell lymphomas. Blood. 130:1081–1088. 2017. View Article : Google Scholar : PubMed/NCBI
34	Dal Col J, Zancai P, Terrin L, Guidoboni M, Ponzoni M, Pavan A, Spina M, Bergamin S, Rizzo S, Tirelli U, et al: Distinct functional significance of Akt and mTOR constitutive activation in mantle cell lymphoma. Blood. 111:5142–5151. 2008. View Article : Google Scholar : PubMed/NCBI
35	Madrid LV, Mayo MW, Reuther JY and Baldwin AS Jr: Akt stimulates the transactivation potential of the RelA/p65 Subunit of NF-kappa B through utilization of the Ikappa B kinase and activation of the mitogen-activated protein kinase p38. J Biol Chem. 276:18934–18940. 2001. View Article : Google Scholar : PubMed/NCBI
36	Ahmad A, Biersack B, Li Y, Kong D, Bao B, Schobert R, Padhye SB and Sarkar FH: Targeted regulation of PI3K/Akt/mTOR/NF-κB signaling by indole compounds and their derivatives: Mechanistic details and biological implications for cancer therapy. Anticancer Agents Med Chem. 13:1002–1013. 2013. View Article : Google Scholar : PubMed/NCBI
37	Tanaka K, Babic I, Nathanson D, Akhavan D, Guo D, Gini B, Dang J, Zhu S, Yang H, De Jesus J, et al: Oncogenic EGFR signaling activates an mTORC2-NF-κB pathway that promotes chemotherapy resistance. Cancer Discov. 1:524–538. 2011. View Article : Google Scholar : PubMed/NCBI
38	Lee K, Gudapati P, Dragovic S, Spencer C, Joyce S, Killeen N, Magnuson MA and Boothby M: Mammalian target of rapamycin protein complex 2 regulates differentiation of Th1 and Th2 cell subsets via distinct signaling pathways. Immunity. 32:743–753. 2010. View Article : Google Scholar : PubMed/NCBI
39	Stewart ZA, Westfall MD and Pietenpol JA: Cell-cycle dysregulation and anticancer therapy. Trends Pharmacol Sci. 24:139–145. 2003. View Article : Google Scholar : PubMed/NCBI
40	Halazonetis TD, Gorgoulis VG and Bartek J: An oncogene-induced DNA damage model for cancer development. Science. 319:1352–1355. 2008. View Article : Google Scholar : PubMed/NCBI
41	Riedl SJ and Salvesen GS: The apoptosome: Signalling platform of cell death. Nat Rev Mol Cell Biol. 8:405–413. 2007. View Article : Google Scholar : PubMed/NCBI
42	LoPiccolo J, Blumenthal GM, Bernstein WB and Dennis PA: Targeting the PI3K/Akt/mTOR pathway: Effective combinations and clinical considerations. Drug Resist Updat. 11:32–50. 2008. View Article : Google Scholar : PubMed/NCBI
43	Sabatini DM: mTOR and cancer: Insights into a complex relationship. Nat Rev Cancer. 6:729–734. 2006. View Article : Google Scholar : PubMed/NCBI
44	Gingras AC, Gygi SP, Raught B, Polakiewicz RD, Abraham RT, Hoekstra MF, Aebersold R and Sonenberg N: Regulation of 4E-BP1 phosphorylation: A novel two-step mechanism. Genes Dev. 13:1422–1437. 1999. View Article : Google Scholar : PubMed/NCBI
45	Hsu PP, Kang SA, Rameseder J, Zhang Y, Ottina KA, Lim D, Peterson TR, Choi Y, Gray NS, Yaffe MB, et al: The mTOR-regulated phosphoproteome reveals a mechanism of mTORC1-mediated inhibition of growth factor signaling. Science. 332:1317–1322. 2011. View Article : Google Scholar : PubMed/NCBI
46	Zhang HT, Wang WW, Ren LH, Zhao XX, Wang ZH, Zhuang DL and Bai YN: The mTORC2/Akt/NFκB pathway-mediated activation of TRPC6 participates in adriamycin-induced podocyte apoptosis. Cell Physiol Biochem. 40:1079–1093. 2016. View Article : Google Scholar
47	Lee K, Nam KT, Cho SH, Gudapati P, Hwang Y, Park DS, Potter R, Chen J, Volanakis E and Boothby M: Vital roles of mTOR complex 2 in Notch-driven thymocyte differentiation and leukemia. J Exp Med. 209:713–728. 2012. View Article : Google Scholar : PubMed/NCBI
48	Bassères DS and Baldwin AS: Nuclear factor-kappaB and inhibitor of kappaB kinase pathways in oncogenic initiation and progression. Oncogene. 25:6817–6830. 2006. View Article : Google Scholar : PubMed/NCBI
49	Karin M: Nuclear factor-kappaB in cancer development and progression. Nature. 441:431–436. 2006. View Article : Google Scholar : PubMed/NCBI
50	Yun S, Vincelette ND, Knorr KL, Almada LL, Schneider PA, Peterson KL, Flatten KS, Dai H, Pratz KW, Hess AD, et al: 4EBP1/c-MYC/PUMA and NF-κB/EGR1/BIM pathways underlie cytotoxicity of mTOR dual inhibitors in malignant lymphoid cells. Blood. 127:2711–2722. 2016. View Article : Google Scholar : PubMed/NCBI