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OBJECTIVE To investigate the inhibitory effects of Spatholobus Suberectus Column Extract(SSCE)on estrogen receptor positive(ER+)breast cancer cel MCF-7and its possible molecular mechanism.METHODS MCF-7cells were cultured without estrogen and with 17-β-estrogen(10-8mol·L-1),respectively,then treated with SSCE(0,40,80,160,320μg·m L-1).MTT assay was employed to evaluate cell viability.Flow cytometry assays were performed to underlying apoptosis and detecting cel cycle of MCF-7 cells treated with SSCE(0,80,160,320μg·mL-1).Wound healing assays was conducted to detect the migration ability.Dual luciferase reporter system was used to detect the activity of p-ERα,p-ERβpresented in intra-nuclear estrogen response element(ERE).Western blotting assay was employed to identify the expression of protein such as Bax,Bcl-2,p-ERα,p-ERβ,ERK1/2,p-ERK1/2,AKT,p-AKT,m TOR,p-m TOR,PI3K,p-PI3K.RESULTS It showed that SSCE(80,160and 320μg·mL-1)significantly decreased the viability of MCF-7.SSCE also triggered apoptosis,arrested cell cycle at G0/G1phase,inhibited migration.Dual luciferase reporter system showed that SSCE suppressed intra-nuclear p-ER activity,Western blotting analysis confirmed that SSCE did repress the expression of phosphorylated-ER alpha(p-ERα),ERK1/2,p-ERK1/2,AKT,p-AKT,pmT OR,PI3K,p-PI3K,which indicate that SSCE suppress MAPK PI3K/AKT signal pathway.CONCLUSION Our result showed that SSCE cause ER+MCF-7 cells apoptosis,G0/G1phase arresting,migration decreasing,via hypo-active of ER,suppress MAPK PI3K/AKT pathway.
OBJECTIVE To investigate the inhibitory effects of Spatholobus Suberectus Column Extract (SSCE) on estrogen receptor positive (ER +) breast cancer cel MCF-7 and its possible molecular mechanism. METHODS MCF-7 cells were cultured without estrogen and with 17-beta-estrogen 8mol·L-1), respectively, then treated with SSCE (0, 40,80,160,320μg · m L-1) .MTT assay was employed to evaluate cell viability. Flow cytometry assays were performed to the underlying apoptosis and detecting cel cycle of MCF -7 cells treated with SSCE (0, 80, 160, 320 μg · mL -1) .Wound healing assays was conducted to detect the migration ability. Dual luciferase reporter system was used to detect the activity of p-ERα, p-ERβpresented in intra-nuclear Western blotting assay was employed to identify the expression of protein such as Bax, Bcl-2, p-ERα, p-ERβ, ERK1 / 2, p-ERK1 / 2, AKT, p-AKT, m TOR, pm TOR, PI3K, p-PI3K.RESULTS It showed that SSCE (80,160 and 320μg · mL-1) significantly decreased the viability of MCF-7.SSCE also triggered apoptosis, arrested cell cycle at G0 / G1phase, inhibited migration. Dual luciferase reporter system showed that SSCE suppressed intra-nuclear p-ER activity, Western blotting analysis confirmed that SSCE did repress the expression of phosphorylated-ER alpha (p-ERα) , ERK1 / 2, p-ERK1 / 2, AKT, p-AKT, pmT OR, PI3K, p-PI3K, which indicates that SSCE suppress MAPK PI3K / AKT signal pathway. CONCLUSION Our result showed that SSCE cause ER + MCF-7 cells apoptosis, G0 / G1phase arresting, migration decreasing, via hypo-active of ER, suppress MAPK PI3K / AKT pathway.