摘要
目的 :研究MEK 1在MCF 7乳腺癌细胞对αV整合素 (αVintegrin)和上皮钙粘素 (E cadherin)调节的表达。方法 :应用细胞转化技术把含有点突变的基因MEK 1整合进MCF 7乳腺癌细胞 ,使之成为含有整合基因MEK 1 GFP、Ala2 2 2 MEK 1 GFP、Asp2 2 2 MEK GFP的MCF 7乳腺癌细胞 ,通过免疫杂交和免疫荧光技术检测其表达水平的变化和在细胞内的定位。结果 :无活性的Ala2 2 2 MEK 1 GFP抑制MAPK的活性 ,增加上皮钙粘素的表达 ;活化的Asp2 2 2 MEK GFP增加MAPK的活性 ,不影响上皮钙粘素的表达 ;MEK 1 GFP和Asp2 2 2 MEK GFP上调αV整合素的表达 ,增加FAK磷酸化 ,Ala2 2 2 MEK 1 GFP不增加αV整合素的表达 ,抑制FAK的磷酸化。结论 :MEK调节αV整合素和上皮钙粘素的表达 ,而FAK是诱导整合素信号通路的重要因子 ,MEK可能通过调节整合素和上皮钙粘素的表达来调节FAK的磷酸化。
Purpose:Expression of MEK-1 was studied by regulating αV integrin and E-cadherin on MCF-7 breast cancer cells.Methods:Using cell transfected techniques site-mutated genes were inserted into MCF-7 cells and made MCF-7 cells containing MEK-1-GFP,Ala 222 MEK-1-GFP,Asp 222 MEK-1-GFP ,respectively. These cells were tested with immunoblot analysis and immunofluoresence technique to determined the change of MEK-1 expression and location of MEK-1 in cells.Results:Deactivated Ala 222 MEK-1-GFP inhibited the activity of MAPK,active Asp 222 MEK-1-GFP increased the activity of MAPK. Ala 222 MEK-1-GFP increased the expression of E-cadherin,did not increase the expression of αV integrin and inhibited FAK phosphorylation. MEK-1-GFP and Asp 222 MEK-1-GFP did not affect the expression of E-cadherin ,but upregulated the expression of αV integrin,increased FAK phosphorylation.Conclusions:MEK-1 regulated the expression of αV integrin and E-cadherin,FAK is the key factor that induced signal pathway of αV integrin. It is possible that MEK-1 regulated FAK phosphorglation by regulating expression of αV integrin and E-cadherin.
出处
《中国癌症杂志》
CAS
CSCD
2003年第6期511-514,共4页
China Oncology
