摘要
目的:研究JAK激酶抑制剂AG490对乳腺癌细胞MDA-MB-231细胞外信号调节蛋白激酶(ERK)磷酸化的影响,探讨JAK激酶对ERK信号转导通路的作用以及在乳腺癌细胞侵袭转移中的调控意义。方法:以JAK激酶抑制剂AG490处理乳腺癌细胞MDA-MB-231,MTT法检测AG490对细胞的生长抑制作用;MTT法检测AG490处理后细胞对人工基底膜Matrigel的黏附能力变化;Transwell小室进行人工重组基底膜侵袭和迁移实验,检测AG490处理后细胞侵袭、迁移能力变化;Western印迹法检测AG490处理后细胞中磷酸化ERK(P-ERK)、基质金属蛋白酶9(MMP-9)蛋白水平变化。结果:应用JAK酶抑制剂AG490后MDA-MB-231细胞生长受到抑制,作用呈时效-量效依赖关系;MDA-MB-231细胞黏附、侵袭、迁移能力降低,同时细胞中P-ERK、MMP-9蛋白减少。结论:JAK激酶可以通过改变ERK的磷酸化水平影响ERK信号转导通路的活化。JAK激酶抑制对ERK信号转导通路的激活有阻断作用,可以抑制MMP-9的表达及乳腺癌细胞的侵袭转移。
Objective：To explore the influence of Janus kinase inhibitor AG490 on phosphorylation of extracellular signal-regulated protein kinase （ERK） in the human breast cancer cell line MDA-MB-231, so as to discuss the effect of JAK on ERK signal transduction pathway and on invasion/metastasis of breast cancer cells. Methods： MDA-MB-231 cells were treated with Janus kinase inhibitor AG490;MTT assay was used to examine the proliferation of MDA-MB-231 cells and the adhesion of MDA MB-231 cells to artificial basement membrane matrigel after AG490 treatment. Invasion and metastasis of MDA-MB-231 cells were evaluated with transwell chamber after treated with AG490. The expression of P ERK and MMP 9 protein was determined by Western blotting assay. Results： AG490 inhibited the proliferation of MDA MB-231 cells in a time- and dose-dependent manner; AG490 also depressed the adhesion, invasion and metastasis of MDA-MB-231 cells. Meanwhile, the expression of P-ERK and MMP 9 protein was decreased after treatment with AG490. Conclusion： Our study indicates that JAK kinase can affect the activity of ERK signal transduction pathway through the phosphorylation of ERK. The inhibitory effects of JAK kinase on MMP-9 expression and invasion of breast cancer cells are associated with the blocking of the ERK signaling pathway.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2008年第8期900-903,共4页
Academic Journal of Second Military Medical University
