Influence of Src kinase inhibitor ZD6474 on breast cancer MCF-7 cells and its mechanism

ZHAO Yu-tao; YANG Ying-hua; ZHAO Jian-ping

doi:10.3969/j.issn.1002-266X.2015.45.005

Abstract：Objective To investigate the effects of Src kinase inhibitor ZD 6474 on the proliferation of human breast cancer MCF-7 cells and its mechanism .Methods MCF-7 cells in logarithmic phase were treated with different concentra-tions of Src kinase inhibitor ZD6474 (1 ×10 -6 mol/L, 1 ×10 -5 mol /L, 1 ×10 -4 mol /L, 1 ×10 -3 mol /L and 1 ×10 -2 mol /L) .The cell growth inhibition rate was calculated by MTT method .Transwell assay was used to detect the invasion a-bility of MCF-7 cells in vitro.Western blotting was used to detect the protein expression of Src , E-cadherin andβ-catenin. The activity of Snail promoter was detected by reporter gene technology .Real-time PCR was used to detect the mRNA ex-pression of E-cadherin and β-catenin.Results When the MCF-7 cells were treated with ZD6474 at the concentrations of 1 ×10 -5 mol/L and 1 ×10 -4 mol/L, the inhibition rates were 12.2% and 27.5%.The invasion ability of MCF-7 cells in vitro was significantly decreased after being treated with ZD 6474 .The invasion abilities of MCF-7 cells treated with 1 × 10 -6 mol/L, 1 ×10 -5 mol/L,1 ×10 -4 mol/L,1 ×10 -3 mol/L and 1 ×10 -2 mol/L ZD6474 were 8.3%, 14.2%, 32.6%, 51.4%and 76.5%, respectively.Src tyrosine kinase inhibitor significantly up-regulated the expression of E-cad-herin at protein and mRNA levels , and down-regulated the expression of β-catenin at protein and mRNA levels as well as promoter activity of Snail .Conclusion Src kinase inhibitor ZD6474 could inhibit the proliferation of breast cancer cells , up-regulate the activity of E-caherin, down-regulate the expression of β-catenin and decrease the activity of Snail promoter , and thus it inhibits the invasion and metastasis of breast cancer cells .