Src 激酶抑制剂 ZD6474 对乳腺癌 MCF-7细胞增殖的影响及机制

摘要

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 .%目的 探讨Src激酶抑制剂ZD6474对乳腺癌MCF-7细胞增殖的影响及机制. 方法 取对数生长期的MCF-7细胞,分别加入1 ×10 -6 mol/L、1 ×10 -5 mol/L、1 ×10 -4 mol/L、1 ×10 -3 mol/L、1 ×10 -2 mol/L的Src激酶抑制剂ZD6474. 采用MTT法测算细胞增殖抑制率. 采用Transwell法检测MCF-7细胞体外侵袭能力. 采用Western blotting法检测E-钙黏素(E-cadherin)及β-连环蛋白(β-catenin)蛋白表达. 采用报告基因技术检测Snail启动子的转录活性. 采用real-time PCR法检测E-cadherin 和β-catenin mRNA表达. 结果 ZD6474 作用浓度为1 ×10 -5 mol/L和1 ×10 -4 mol/L时,细胞增殖抑制率分别为12 .2%和27 .5%. MCF-7细胞经ZD6474处理后,体外侵袭能力明显下降,1 ×10 -6 mol /L、1 ×10 -5 mol/L、1 ×10 -4 mol/L、1 ×10 -3 mol/L、1 ×10 -2 mol /LZD6474作用MCF-7细胞体外侵袭能力分别为8.3%、14.2%、32.6%、51.4%、76.5%. Src激酶抑制剂作用后E-cadherin mRNA和蛋白表达上调,β-catenin mRNA和蛋白表达及Snail启动子转录活性下调. 结论 Src蛋白激酶抑制剂ZD6474可抑制乳腺癌细胞增殖,其机制可能通过上调E-caherin表达,下调β-catenin表达,减弱Snail启动子转录活性,从而抑制乳腺癌细胞的侵袭和转移.