The dual PI3K/mTOR inhibitor NVP-BEZ235 inhibits proliferation and induces apoptosis of burkitt lymphoma cells

Cancer Cell International

Xiongpeng Zhu¹  Yuanling Huang¹  Yan Zheng¹  Huifang Xiao¹  Pengliang Xin¹  Chuntuan Li¹ 

[1]Department of Haematology, First Hospital of Quanzhou Affiliated to Fujian Medical University, 248 East Street, Licheng District, Quanzhou 362000, Fujian Province, China

关键词: Apoptosis;    Cell proliferation;    NVP-BEZ235;    Phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin pathway;    Burkitt lymphoma;   

Others  :  1219328
DOI  :  10.1186/s12935-015-0213-1

 received in 2015-04-07, accepted in 2015-06-03,  发布年份 2015

【 摘 要 】

Background

Phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway is a therapy target of cancer. We aimed to confirm the effect of dual PI3K/mTOR inhibitor NVP-BEZ235 on cell proliferation and apoptosis in Burkitt lymphoma (BL) cells.

Methods

Two human BL cell lines, CA46 and RAJI were used in this study. The proliferation of BL cells was detected by manganese tricarbonyl transfer (MTT) assay. Cell cycle and apoptosis assay were examined by flow cytometric analysis. The phosphorylation levels of AKT (Thr308), AKT (Ser473), and RPS6 were evaluated by western blot analysis.

Results

NVP-BEZ235 significantly inhibited the proliferation of BL cells (CA46 and RAJI) and the inhibition effect was time and dose-dependent. Cell cycle analysis indicated that the cells (CA46 and RAJI) were mostly arrested in G1/G0 phase. Cell apoptosis assay showed that the late apoptotic cells were significantly increased after 72 h treatment by 100 nmol/L of NVP-BEZ235. In addition, results also found that NVP-BEZ235 reduced the phosphorylation levels of AKT (Thr308), AKT (Ser473), and PRS6 in BL cells (CA46 and RAJI). Moreover, this inhibition effect on phosphorylation was dose-dependent.

Conclusions

NVP-BEZ235 effectively inhibited cell proliferation by G0/G1 cell-cycle arrest and induced apoptosis through deregulating PI3K/Akt/mTOR pathway in BL cells.

【 授权许可】

   
2015 Li et al.

【 图 表 】

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【 参考文献 】

• [1]Leoncini L, Stein H. Definition of Burkitt Lymphoma. In: Burkitt’s Lymphoma. Springer, New York; 2013: p.81-93.
• [2]Perkins AS, Friedberg JW. Burkitt lymphoma in adults. ASH Education Program Book. 2008; 2008:341-8.
• [3]Sekiguchi N, Nishimoto J, Tanosaki R, Kubota N, Yokota Y, Kobayashi Y et al.. EBV-positive Burkitt lymphoma as a late-onset posttransplantion lymphoproliferative disorder after allogeneic stem cell transplantation. Int J Hematol. 2004; 79:387-9.
• [4]Molyneux EM, Rochford R, Griffin B, Newton R, Jackson G, Menon G et al.. Burkitt’s lymphoma. Lancet. 2012; 379:1234-44.
• [5]Mutalima N, Molyneux E, Jaffe H, Kamiza S, Borgstein E, Mkandawire N et al.. Associations between Burkitt lymphoma among children in Malawi and infection with HIV, EBV and malaria: results from a case–control study. PloS ONE. 2008; 3: Article ID e2505
• [6]Cairo MS, Gerrard M, Sposto R, Auperin A, Pinkerton CR, Michon J et al.. Results of a randomized international study of high-risk central nervous system B non-Hodgkin lymphoma and B acute lymphoblastic leukemia in children and adolescents. Blood. 2007; 109:2736-43.
• [7]Mbulaiteye SM, Anderson WF, Ferlay J, Bhatia K, Chang C, Rosenberg PS et al.. Pediatric, elderly, and emerging adult-onset peaks in Burkitt’s lymphoma incidence diagnosed in four continents, excluding Africa. Am J Hematol. 2012; 87:573-8.
• [8]Aka P, Kawira E, Masalu N, Emmanuel B, Brubaker G, Magatti J et al.. Incidence and trends in Burkitt lymphoma in northern Tanzania from 2000 to 2009. Pediatr Blood Cancer. 2012; 59:1234-8.
• [9]Mbulaiteye SM, Clarke CA, Morton LM, Gibson TM, Pawlish K, Weisenburger DD et al.. Burkitt lymphoma risk in US solid organ transplant recipients. Am J Hematol. 2013; 88:245-50.
• [10]Richter-Larrea JA, Robles EF, Fresquet V, Beltran E, Rullan AJ, Agirre X et al.. Reversion of epigenetically mediated BIM silencing overcomes chemoresistance in Burkitt lymphoma. Blood. 2010; 116:2531-42.
• [11]Sander S, Rajewsky K. Burkitt lymphomagenesis linked to MYC plus PI3K in germinal center B cells. Oncotarget. 2012; 3:1066-7.
• [12]Sharp ZD, Bartke A. Evidence for down-regulation of phosphoinositide 3-kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR)-dependent translation regulatory signaling pathways in Ames dwarf mice. J Gerontol A Biol Sci Med Sci. 2005; 60:293-300.
• [13]Morgensztern D, Mcleod HL. PI3K/Akt/mTOR pathway as a target for cancer therapy. Anticancer Drugs. 2005; 16:797-803.
• [14]Brown RE. Morphoproteomics: exposing protein circuitries in tumors to identify potential therapeutic targets in cancer patients. Expert Rev Proteomics. 2005; 2(3):337-48.
• [15]Yip PY. Phosphatidylinositol 3-kinase-AKT-mammalian target of rapamycin (PI3K-Akt-mTOR) signaling pathway in non-small cell lung cancer. Translational Lung Cancer Research. 2015; 4(2):165.
• [16]Domina AM, Van Doorn C-C. The effects of (−)-epigallocatechin-3-gallate (EGCG) and phosphatidylinositol 3-kinase (PI (3) K) inhibition on Burkitt lymphoma cells. FASEB J. 2013; 27:1167.1163.
• [17]Huang Y, Hu J, Zheng J, Li J, Wei T, Zheng Z et al.. Down-regulation of the PI3K/Akt signaling pathway and induction of apoptosis in CA46 Burkitt lymphoma cells by baicalin. J Exp Clin Cancer Res. 2012; 31:48. BioMed Central Full Text
• [18]Skomra A, Czuczman NM, Mavis C, Rolland DC, Lim M, Awasthi A et al.. The Acquirement Of Rituximab Resistance Is Associated With De-Regulation Of The PI3K/Akt/mTOR Signaling Pathway Leading To Chemotherapy Resistance In Burkitt Lymphoma Pre-Clinical Models. Blood. 2013; 122:5144.
• [19]Maira S-M, Stauffer F, Brueggen J, Furet P, Schnell C, Fritsch C 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. 2008; 7:1851-63.
• [20]Mukherjee B, Tomimatsu N, Amancherla K, Camacho CV, Pichamoorthy N, Burma S. The dual PI3K/mTOR inhibitor NVP-BEZ235 is a potent inhibitor of ATM-and DNA-PKCs-mediated DNA damage responses. Neoplasia. 2012; 14:34-IN38.
• [21]Del Alcazar CRG, Hardebeck MC, Mukherjee B, Tomimatsu N, Gao X, Yan J et al.. Inhibition of DNA double-strand break repair by the dual PI3K/mTOR inhibitor NVP-BEZ235 as a strategy for radiosensitization of glioblastoma. Clin Cancer Res. 2014; 20:1235-48.
• [22]Cerniglia GJ, Karar J, Tyagi S, Christofidou-Solomidou M, Rengan R, Koumenis C et al.. Inhibition of autophagy as a strategy to augment radiosensitization by the dual phosphatidylinositol 3-kinase/mammalian target of rapamycin inhibitor NVP-BEZ235. Mol Pharmacol. 2012; 82:1230-40.
• [23]Shortt J, Martin BP, Newbold A, Hannan KM, Devlin JR, Baker AJ et al.. Combined inhibition of PI3K-related DNA damage response kinases and mTORC1 induces apoptosis in MYC-driven B-cell lymphomas. Blood. 2013; 121:2964-74.
• [24]Baumann P, Mandl-Weber S, Oduncu F, Schmidmaier R. The novel orally bioavailable inhibitor of phosphoinositol-3-kinase and mammalian target of rapamycin, NVP-BEZ235, inhibits growth and proliferation in multiple myeloma. Exp Cell Res. 2009; 315:485-97.
• [25]Shoji K, Oda K, Kashiyama T, Ikeda Y, Nakagawa S, Sone K et al.. Genotype-dependent efficacy of a dual PI3K/mTOR inhibitor, NVP-BEZ235, and an mTOR inhibitor, RAD001, in endometrial carcinomas. PloS ONE. 2012; 7: Article ID e37431
• [26]Venkannagari S, Fiskus W, Peth K, Atadja P, Hidalgo M, Maitra A et al.. Superior efficacy of co-treatment with dual PI3K/mTOR inhibitor NVP-BEZ235 and pan-histone deacetylase inhibitor against human pancreatic cancer. Oncotarget. 2012; 3:1416-27.
• [27]Lin S-F, Huang Y-Y, Lin J-D, Chou T-C, Hsueh C, Wong RJ. Utility of a PI3K/mTOR inhibitor (NVP-BEZ235) for thyroid cancer therapy. PloS ONE. 2012; 7: Article ID e46726
• [28]Gobin B, Battaglia S, Lanel R, Chesneau J, Amiaud J, Rédini F et al.. NVP-BEZ235, a dual PI3K/mTOR inhibitor, inhibits osteosarcoma cell proliferation and tumor development in vivo with an improved survival rate. Cancer Lett. 2014; 344:291-8.
• [29]Pajic A, Spitkovsky D, Christoph B, Kempkes B, Schuhmacher M, Staege MS et al.. Cell cycle activation by c-myc in a Burkitt lymphoma model cell line. Int J Cancer. 2000; 87:787-93.
• [30]Muise-Helmericks RC, Grimes HL, Bellacosa A, Malstrom SE, Tsichlis PN, Rosen N. Cyclin D expression is controlled post-transcriptionally via a phosphatidylinositol 3-kinase/Akt-dependent pathway. J Biol Chem. 1998; 273:29864-72.
• [31]Bouchard C, Marquardt J, Bras A, Medema RH, Eilers M. Myc-induced proliferation and transformation require Akt-mediated phosphorylation of FoxO proteins. EMBO J. 2004; 23:2830-40.
• [32]Schmidt M, De Mattos SF, Van Der Horst A, Klompmaker R, Kops GJL, Lam EW-F et al.. Cell cycle inhibition by FoxO forkhead transcription factors involves downregulation of cyclin D. Mol Cell Biol. 2002; 22:7842-52.
• [33]Wan X, Helman LJ. Effect of insulin-like growth factor II on protecting myoblast cells against cisplatin-induced apoptosis through p70 S6 kinase pathway. Neoplasia. 2002; 4:400-8.
• [34]Padmanabhan J, Brown KR, Padilla A, Shelanski ML. Functional role of RNA polymerase II and P70 S6 kinase in KCl withdrawal-induced cerebellar granule neuron apoptosis. J Biol Chem. 2015; 290(9):5267-79.
• [35]Dhar R, Persaud SD, Mireles JR, Basu A. Proteolytic Cleavage of p70 Ribosomal S6 Kinase by Caspase-3 during DNA Damage-Induced Apoptosis†. Biochemistry. 2009; 48:1474-80.
• [36]Sarbassov DD, Guertin DA, Ali SM, Sabatini DM. Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex. Science. 2005; 307:1098-101.
• [37]Brian M, Bilgen E, Diane CF. Regulation and function of ribosomal protein S6 kinase (S6K) within mTOR signalling networks. Biochem J. 2012; 441:1-21.
• [38]Kim JH, Jeong SJ, Kim B, Yun SM, Choi DY, Kim SH. Melatonin synergistically enhances cisplatin‐induced apoptosis via the dephosphorylation of ERK/p90 ribosomal S6 kinase/heat shock protein 27 in SK‐OV‐3 cells. J Pineal Res. 2012; 52:244-52.
• [39]Neise D, Sohn D, Stefanski A, Goto H, Inagaki M, Wesselborg S et al.. The p90 ribosomal S6 kinase (RSK) inhibitor BI-D1870 prevents gamma irradiation-induced apoptosis and mediates senescence via RSK-and p53-independent accumulation of p21WAF1/CIP1. Cell Death Dis. 2013; 4:e859.