ABSTRACT
OBJECTIVE@#To investigate the effect of metformin on the proliferation and apoptosis of HER-2-positive breast cancer cell line SKBR3 and explore the possible mechanism of its action.@*METHODS@#SKBR3 cells were treated with different concentrations (20-120 μmol/L) of metformin, and the changes in cell proliferation and colony formation ability were assessed using CCK-8 assay and crystal violet staining, respectively. Flow cytometry was performed to analyze cell apoptosis and cell cycle changes. Real-time fluorescent quantitative PCR (qRT-PCR) was used to detect mRNA expressions of YAP, TAZ, EGFR, CTGF, CYR61, E-cadherin, N-cadherin, vimentin and fibronectin in the treated cells, and the protein expressions of YAP and TAZ were detected using Western blotting; immunofluorescence assay was used to observe YAP/TAZ nuclear translocation in the cells.@*RESULTS@#Metformin treatment significantly inhibited the proliferation of SKBR3 cells (P < 0.05) in a concentration- and time-dependent manner. The results of flow cytometry showed that metformin significantly promoted apoptosis and caused cell cycle arrest at G1 phase in SKBR3 cells. Metformin treatment significantly down-regulated the mRNA expressions of YAP, TAZ, EGFR, CTGF and CYR61, N-cadherin, vimentin and fibronectin (P < 0.05) and up-regulated the expression of E-cadherin (P < 0.05); Western blotting results showed that YAP and TAZ protein expressions were significantly down-regulated in the cells after metformin treatment (P < 0.05). Immunofluorescence assay revealed that metformin treatment caused the concentration of YAP and TAZ in the cytoplasm, and significantly reduced their amount in the cell nucleus.@*CONCLUSION@#Metformin can inhibit proliferation and promote apoptosis and epithelal-mesenchymal transition of HER-2 positive breast cancer cells possibly by that inhibing YAP and TAZ expression and their nuclear localization.
Subject(s)
Apoptosis , Cadherins , Cell Proliferation , ErbB Receptors , Fibronectins , Metformin/pharmacology , Neoplasms , Protein Serine-Threonine Kinases , RNA, Messenger , Transcription Factors/metabolism , VimentinABSTRACT
Objective To observe the effect of curcumin on proliferation, apoptosis, Caspase-3 activity and telomerase activity of chronic granulocytic breast cancer SKBR3 cell line, and to investigate the mechanism of curcumin. Methods The cell growth curve was drawn by MTT;the apoptotic cells were detected by Annexin, V and PI double staining; the expression of Caspase-3 was detected by Westernblot; the activity of telomerase was detected by TRAP-PCR silver staining. Results Curcumin could significantly inhibit the proliferation of SKBR3 cells in a time and concentration dependent, has a certain effect on reducing the telomerase activity of SKBR3 cells can enhance the expression of Caspase-3 induced apoptosis. Conclusion Curcumin can effectively inhibit the proliferation of SKBR3 cells, increase the synthesis of Caspase-3 protein, inhibit the activity of telomerase and mediate the apoptosis of SKBR3 cells.
ABSTRACT
Objective To observe the effect of curcumin on proliferation, apoptosis, Caspase-3 activity and telomerase activity of chronic granulocytic breast cancer SKBR3 cell line, and to investigate the mechanism of curcumin. Methods The cell growth curve was drawn by MTT;the apoptotic cells were detected by Annexin, V and PI double staining; the expression of Caspase-3 was detected by Westernblot; the activity of telomerase was detected by TRAP-PCR silver staining. Results Curcumin could significantly inhibit the proliferation of SKBR3 cells in a time and concentration dependent, has a certain effect on reducing the telomerase activity of SKBR3 cells can enhance the expression of Caspase-3 induced apoptosis. Conclusion Curcumin can effectively inhibit the proliferation of SKBR3 cells, increase the synthesis of Caspase-3 protein, inhibit the activity of telomerase and mediate the apoptosis of SKBR3 cells.