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Objective: To investigate the effects of β-lapachone combined with BEZ235 on the proliferation and migration of breast cancer cells, and to explore its possible mechanism. Methods: The effects of β-lapachone and BEZ235 alone or combination on the proliferation of breast cancer MCF-7 and MDA-MB-468 cells were detected by MTT assay, and the combination index (CI) was calculated. The expression level of proliferation-related protein Ki67 in MCF-7 and MDA-MB-468 cells was detected by Western blotting. The colony formation ability of MCF-7 and MDA-MB-468 cells was detected by plate cloning assay. The lateral migration, vertical migration and invasion abilities of MCF-7 and MDA-MB-468 cells were detected by scratch healing experiment and Transwell chamber assay, respectively. The expression levels of epithelialmesenchymal transition (EMT)-related proteins in MCF-7 and MDA-MB-468 cells were detected by Western blotting. The angiogenesis assay was used to detect the angiogenic ability of human umbilical vein endothelial cells (HUVECs) cultured with the supernatants of MCF-7 and MDAMB- 468 cells treated with β-lapachone, BEZ235 and their combination. The effects of β-lapachone and BEZ235 alone or combination on the expression of vascular endothelial growth factor (VEGF) in breast cancer MCF-7 and MDA-MB-468 cells were detected by Western blotting. Results: The proliferation abilities of breast cancer MCF-7 and MDA-MB-468 cells in β-lapachone, BEZ235 alone or combination treatment groups were lower than those in the control group (without any treatment) (all P < 0.05). The two drugs had synergistic effect with CI < 1. The expression level of Ki67 protein in MCF-7 and MDA-MB-468 cells in β-lapachone combined with BEZ235 group was lower than that in the control and single drug groups (all P < 0.05). The number of colony formation of MCF-7 and MDA-MB-468 cells in β-lapachone combined with BEZ235 group was less than that in the control and single drug groups (all P < 0.05). The abilities of lateral migration, vertical migration and invasion of MCF-7 and MDA-MB-468 cells in β-lapachone combined with BEZ235 group were lower than those in the control and single drug groups (all P < 0.05). The expression level of EMTrelated protein E-cadherin in MCF-7 and MDA-MB-468 cells treated with β-lapachone and BEZ235 was higher than that in the control and single drug groups (all P < 0.05), whereas the expression levels of Vimentin and Twist were opposite (all P < 0.05). The vascularization number of HUVECs, which cultured in the supernatants of MCF-7 and MDA-MB-468 cells treated with β-lapachone and BEZ235, was significantly lower than that in the control and single drug groups (all P < 0.01). The expression level of VEGF in MCF-7 and MDA-MB-468 cells treated with β-lapachone and BEZ235 was lower than that in the control and single drug groups (all P < 0.05). Conclusion: Combination of β-lapachone and BEZ235 can synergistically inhibit the proliferation of breast cancer cells in vitro, and suppress cell invasion and metastasis through angiogenesis and EMT pathway.
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Aim To evaluate the efficacy of NVP-BEZ235 on the proliferation and migration of HepG2 human hepatocellular carcinoma cell in vitro, and to investigate the molecular mechanism. Methods The cytotoxicity of NVP-BEZ235 on HepG2 cells treated with NVP-BEZ235(0, 25, 50, 100, 250, 500 nmol·L-1) was detected by MTT assay. The ability of NVP-BEZ235 to modulate HepG2 cells proliferation and apoptosis was detected by morphology assay and Hoechst 33342 staining. The motility of NVP-BEZ235 treatment on HepG2 cells was determined by wound healing assay and Transwell assay.The expression levels of the related protein including PI3K/Akt/mTOR signal pathway, epithelial-mesenchymal transition(EMT) process and Six1/Ezrin signal axis were detected by Western blot. The expression levels of EMT markers were also detected by immunofluorescence staining. Results The cell viability significantly decreased in HepG2 cells treated with NVP-BEZ235 compared with control group, which showed a dose-dependent manner. NVP-BEZ235 could induce apoptosis and inhibit HepG2 cell migration. E-cadherin protein expression significantly increased; however, the expressions of p-AktSer473, p-S6Thr389, p-4E-BP1Thr37/46, Vimentin, Snail, Six1 and p-EzrinTyr353 decreased in NVP-BEZ235 treated HepG2 cells compared with those in control group. Conclusions NVP-BEZ235 could effectively suppress the proliferation and migration of HepG2 cells, which may be related to the down-regulation of p-AktSer473, p-S6Thr389, p-4E-BP1Thr37/46 and modulating both Six1/Ezrin signal axis and the EMT process by NVP-BEZ235 treatment.
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NVP-BEZ235 is a newly developed dual phosphatidyl linositol kinase 3-kinase (PI3K) and mammalian target of rapamycin (mTOR) inhibitor.Studies have revealed that NVP-BEZ235 combinate with other drugs were effective in the treatment of lung cancer.NVP-BEZ235 might have a potential application value for the treatment of lung cancer of EGFR mutation subtype and KRAS mutation subtype.
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Objective To investigate the effects of NVP-BEZ235 on proliferation and apoptosis of human cholangiocarcinoma(CCA) cell QBC939 in vitro and to reveal the antineoplastic mechanisms of NVP-BEZ235.Methods Human CCA cell line QBC939 was used in this study.Cell apoptosis by NVP-BEZ235 was analyzed using the flow cytometry.Cell growth inhibition by NVP-BEZ235 for 24h 48h 72h by MTT assay.The antineoplastic mechanisms of NVP-BEZ235 for 48h were assessed by western blotting for PARP、Bcl-2、Akt、p-Akt、c-FLIPL and Mcl-1 assay in QBC939 cell.Results NVP-BEZ235 treatment inhibited the proliferation and induced apoptosis of human CCA cell QBC939,which appeared time-dependent and concentration-dependent effects.NVP-BEZ235 reduced protein levels of Mcl-1、c-FLIPL and Bcl-2 and downregulate protein level of p-Akt significantly.Conclusion NVP-BEZ235 inhibited the phosphorylation of Akt.NVP-BEZ235 downregulated Bcl-2、c-FLIPL、Mcl-1 protein level via PI3K/AKT signaling pathway to induce apoptosis and inhibit the proliferation of human CCA cell QBC939.
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Objective:To determine effects of BEZ235,an inhibitor of phosphoionsitol-3-kinase (PI3K)/ mTOR,on the cell proliferation and migration in human prostate carcinoma lines including RWPE-1,PC3,and DU 145 cells.Methods:Viability of RWPE-1,PC3,and DU145 cells was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay,while cell migration was analyzed by wound healing assay.Western blot and immunofluorescence were used to examine the changes of relevant protein expression.Results:The proliferation of PC3 and DU145 cells was effectively inhibited by BEZ235 (P<0.01),whereas RWPE-1 was not obviously inhibited.Invasion and migration of PC3 and DU145 cells were attenuated by BEZ235 via EMT pathway.Conclusion:The PI3K/mTOR dual inhibitor BEZ235 shows substantial anti-tumor activity in human prostate carcinoma lines of PC3 and DU145 cells,which may be involved in the EMT pathway.