Combination of β-lapachone and BEZ235 inhibits proliferation and migration of breast cancer cells / 肿瘤

ABSTRACT

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.