伏立诺他协同PI3K抑制剂NVP-BEZ235诱导Jurkat细胞凋亡
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摘要
目的探讨伏立诺他联合PI3K抑制剂NVP-BEZ235对Jurkat细胞凋亡的影响,并探讨其作用机制。方法体外培养Jurkat细胞,用不同浓度伏立诺他或(和)NVP-BEZ235孵育后,采用MTS观察两者单独用药或联合用药对细胞的增殖的影响,流式细胞术检测细胞凋亡;Western blot检测PI3K/Akt磷酸化及Bim1表达。结果0.12、0.24、0.36、0.48和0.60μmol/L伏立诺他和6、12、18、24、30 nmol/L NVP-BEZ235对Jurkat细胞的生长增殖具有协同作用。伏立诺他或(和)NVP-BEZ235也可促进Jurkat细胞凋亡,并抑制PI3K/Akt磷酸化,上调Bim1的表达。结论伏立诺他对PI3K抑制剂NVP-BEZ235抑制Jurkat细胞生长具有协同作用,并能诱导Jurkat细胞凋亡,其机制与影响Bim1有关。
Objective To investigate the effect of vorinostat combined with PI3 K inhibitor NVP-BEZ235 on apoptosis of Jurkat cells and to explore its mechanism.Methods Jurkat cells were cultured in vitro,and after incubation with different concentrations of vorinostat or(and) NVP-BEZ235,MTS was used to observe the effect of the two drugs alone or in combination on cell proliferation.And apoptosis was detected by flow cytometry.The PI3 K/Akt phosphorylation and Bim1 expression were detected by Western blot.Results The 0.12,0.24,0.36,0.48 and 0.60 μmol/L vorinostat and 6,12,18,24,30 nmol/L NVP-BEZ235 had synergistic effects on the growth and proliferation of Jurkat cells.Vorinostat or(and) NVP-BEZ235 also promoted Jurkat cell apoptosis and inhibited PI3 K/Akt phosphorylation,up-regulated Bim1 expression.Conclusion Vorinostat has a synergistic effect on the inhibition of Jurkat cell growth by PI3 K inhibitor NVP-BEZ235,and can induce apoptosis of Jurkat cells.The mechanism is related to the influence of Bim1.
Objective To investigate the effect of vorinostat combined with PI3 K inhibitor NVP-BEZ235 on apoptosis of Jurkat cells and to explore its mechanism.Methods Jurkat cells were cultured in vitro,and after incubation with different concentrations of vorinostat or(and) NVP-BEZ235,MTS was used to observe the effect of the two drugs alone or in combination on cell proliferation.And apoptosis was detected by flow cytometry.The PI3 K/Akt phosphorylation and Bim1 expression were detected by Western blot.Results The 0.12,0.24,0.36,0.48 and 0.60 μmol/L vorinostat and 6,12,18,24,30 nmol/L NVP-BEZ235 had synergistic effects on the growth and proliferation of Jurkat cells.Vorinostat or(and) NVP-BEZ235 also promoted Jurkat cell apoptosis and inhibited PI3 K/Akt phosphorylation,up-regulated Bim1 expression.Conclusion Vorinostat has a synergistic effect on the inhibition of Jurkat cell growth by PI3 K inhibitor NVP-BEZ235,and can induce apoptosis of Jurkat cells.The mechanism is related to the influence of Bim1.
引文
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[13] Sunters A,de Mattos SF,Stahl M,et al. FoxO3a transcriptional regulation of Bim controls apoptosis in paclitaxel-treated breast cancer cell lines[J]. J Biol Chem,2003,278(50):49795-49805.
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[15] Lee J,Bartholomeusz C,Mansour O,et al. A classⅠhistone deacetylase inhibitor,entinostat,enhances lapatinib efficacy in HER2-overexpressing breast cancer cells through FOXO3-mediated Bim1 expression[J]. Breast Cancer Res Treat,2014,146(2):259-272.
[2] Choi BH,Kim CG,Lim Y,et al. Curcumin down-regulates the multidrug-resistance mdr1b gene by inhibiting the PI3K/Akt/NF kappa B pathway[J]. Cancer Lett,2008,259(1):111-118,
[3] Pereira JK,Machado-Neto JA,Lopes MR,et al.Molecular effects of the phosphatidylinositol-3-kinase inhibitor NVP-BKM120 on T and B-cell acute lymphoblastic leukaemia[J]. Eur J Cancer,2015,51(14):2076-2085.
[4] Ohta H,Aoyagi K,Fukaya M,et al.Cross talk between hedgehog and epithelial-mesenchymal transition pathways in gastric pit cells and in diffuse-type gastric cancers[J]. Br J Cancer,2009,100(2):389-398.
[5] Dummer R,Beyer M,Hymes K,et al.Vorinostat combined with bexarotene for treatment of cutaneous T-cell lymphoma:In vitro and phase I clinical evidence supporting augmentation of retinoic acid receptor/retinoid X receptor activation by histone deacetylase inhibition[J].Leuk Lymphoma,2012,53(8):1501-1508.
[6] Chou TC,Talalay P. Quantitative analysis of dose-effect relationships:The combined effects of multiple drugs or enzyme inhibitors[J]. Adv Enzyme Regul,1984,22:27-55.
[7] Morishita N,Tsukahara H,Chayama K,et al,Activation of Akt is associated with poor prognosis and chemotherapeutic resistance in pediatric B-precursor acute lymphoblastic leukemia[J]. Pediatr Blood Cancer,2012,59(1):83-89.
[8] Baumann P,Mandl-Weber S,Oduncu F,et al. The novel orally bioavailable inhibitor of phosphoinositol-3-kinase and mammalian target of rapamycin,NVP-BEZ235,inhibits growth and proliferation in multiple myeloma[J].Exp Cell Res,2009,315(3):485-497.
[9] Schult C,Dahlhaus M,Glass A,et al. The dual kinase inhibitor NVP-BEZ235 in combination with cytotoxic drugs exerts anti-proliferative activity towards acute lymphoblastic leukemia cells[J]. Anticancer Res,2012,32(2):463-474.
[10] Kim A,Park S,Lee JE,et al,The dual PI3K and mTOR inhibitor NVP-BEZ235 exhibits anti-proliferative activity and overcomes bortezomib resistance in mantle cell lymphoma cells[J]. Leuk Res,2012,36(7):912-920.
[11] Zhao L,Au JL,Wientjes MG. Comparison of methods for evaluating drug-drug interaction[J]. Front Biosci(Elite Ed),2010,2:241-249.
[12] Gillings AS,Balmanno K,Wiggins CM,et al. Apoptosis and autophagy:BIM as a mediator of tumour cell death in response to oncogene-targeted therapeutics[J]. FEBS J,2009,276(21):6050-6062.
[13] Sunters A,de Mattos SF,Stahl M,et al. FoxO3a transcriptional regulation of Bim controls apoptosis in paclitaxel-treated breast cancer cell lines[J]. J Biol Chem,2003,278(50):49795-49805.
[14] Ajabnoor GM,Crook T,Coley HM. Paclitaxel resis tance is associated with switch from apoptotic to autophagic cell death in MCF-7 breast cancer cells[J]. Cell Death Dis,2012,3:e260.
[15] Lee J,Bartholomeusz C,Mansour O,et al. A classⅠhistone deacetylase inhibitor,entinostat,enhances lapatinib efficacy in HER2-overexpressing breast cancer cells through FOXO3-mediated Bim1 expression[J]. Breast Cancer Res Treat,2014,146(2):259-272.