酪氨酸激酶抑制剂tyrphostin AG1478对乳腺癌细胞作用的机制探讨  

The action mechanism underlying the effect of epidermal growth factor receptor inhibitor tyrphostin AG1478 on human breast cancer cells

作　　者：李萍[1] 傅深[1] 章青[1] 俞立萍[1] 徐文才[1] 孙宜[1] 

机构地区：[1]上海交通大学附属第六人民医院肿瘤放疗科,上海200233

出　　处：《肿瘤》2010年第6期481-485,共5页Tumor

基　　金：上海市科学技术委员会科研计划项目(编号:08411960600;09411951100)

摘　　要：目的:探讨表皮生长因子受体(epithelial growth factor receptor,EGFR)的小分子酪氨酸激酶抑制剂tyrphostin AG1478对乳腺癌细胞的作用机制。方法:以乳腺癌细胞MCF-7(EGFR-/+)和MDA-MB-468(EGFR+++)为研究对象,采用MTT及克隆形成实验检测AG1478对乳腺癌MCF-7和MDA-MB-468细胞增殖和生长效应的影响;Western印迹法测定EGFR、Akt和丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)的表达及活化水平;进一步应用FCM检测AG1478对MDA-MB-468细胞早期凋亡的影响。结果:AG1478可抑制MDA-MB-468细胞的增殖和生长(P<0.05),对MCF-7细胞增殖则无明显的抑制效应(P>0.05);同时,MDA-MB-468细胞经AG1478处理后,细胞中磷酸化Akt(p-Akt)和磷酸化MAPK(p-MAPK)的表达水平明显降低(P<0.05);FCM检测结果表明,AG1478可增加MDA-MB-468细胞的早期凋亡水平(P<0.05)。结论:AG1478可下调PI3K/Akt及Ras/Raf/MAPK信号通路,从而抑制乳腺癌MDA-MB-468细胞增殖及生长,并促进细胞凋亡;但其对MCF-7细胞无明显作用。Objective：To investigate the effect of small-molecule tyrosine kinase inhibitor of epidermal growth factor receptor （EGFR）, tyrphostin AG1478, on human breast cancer cells in vitro and elucidate the action mechanism.Methods：Human breast cancer MCF-7 （EGFR-/＋） and MDA-MB-468 （EGFR＋＋＋） cell lines were used in this study. Cell proliferation and growth capacity were assayed by MTT assay and colony formation assay. The expression and activation of EGFR and the downstream proteins Akt and mitogen-activated protein kinase （MAPK） were detected by Western blotting. The effect of AG1478 on apoptosis of MDA-MB-468 cells was detected by flow cytometry.Results：The MTT and colony formation assay showed that the AG1478 significantly inhibited the proliferation and growth of MDA-MB-468 cells compared with the control group （P〈0.05）, but had no significant effect on MCF-7 cells （P〈0.05）. The expressions of p-Akt and p-MAPK were down-regulated in MDA-MB-468 cells after treatment with AG1478 （P〈0.05）. Flow cytometry analysis showed that AG1478 increased the apoptotic rate of MDA-MB-468 at early stage （P〈0.05）. Conclusion：AG1478 inhibits the proliferation and growth and induced apoptosis of breast cancer MDA-MB-468 cells by down-regulation of PI3K/Akt and Ras/Raf/MAPK signaling pathway. But it had no effects on MCF-7 cells.

关 键 词：乳腺肿瘤 受体 表皮生长因子 蛋白酶抑制药 细胞 培养的 TYRPHOSTIN AG1478 

分 类 号：R737.9[医药卫生—肿瘤]