摘要
目的研究表皮生长因子 (EGF)在子宫内膜癌细胞信号转导中 ,激活细胞外信号调节激酶 (ERK)的情况 ,探讨抑癌蛋白PTEN在EGF诱导的对ERK的活化作用。方法构建野生型PTEN和突变型PTEN(G12 9E)编码基因的逆转录病毒载体 ,体外转染子宫内膜癌细胞Ishikawa ,用G4 18筛选稳定表达的细胞系。用Westernblot检测PTEN基因在Ishikawa细胞中的表达 ,以及EGF诱导Ishikawa EGFP、Ishikawa PTEN和Ishikawa PTEN(G12 9E)中活化ERK的浓度效应和时相特点。结果EGF可激活ERK ,但以激活ERK2为主。以不同浓度的EGF作用于Ishikawa EGFP和Ishikawa PTEN时 ,10 μg/L的EGF可使ERK达到最大活化 ,刺激 5min活化最明显。与Ishikawa EGFP相比较 ,EGF对Ishikawa PTEN中ERK的活化作用明显减弱 ,但未降低到阴性对照的水平。 10 μg/L的EGF刺激Ishikawa PTEN(G12 9E) 5min ,与Ishikawa EGFP相比较无明显改变。结论EGF可刺激子宫内膜癌细胞ERK活化 。
Aim To study the activation of extracellular signal regulated kinase(ERK) induced by EGF on endometrial carcinoma cell line Ishikawa and to explore the role of suppressor oncoprotein PTEN in activation of ERKs. Methods Wild type PTEN and mutant PTENG (G129E) gene were subcloned into retroviral vector pLTSN and endometrial carcinoma cell line Ishikawa was transfected. Stable expression of suppressor oncoproteins in Ishikawa were screened by G418. Optimal concentration and induction time of EGF which activated ERKs in Ishikawa PTEN and Ishikawa PTEN (G129E) cells were detected by Western blot. Results EGF could activate ERKs. The activation of ERK reached maximal level at 5 min after stimulation with EGF of 10 μg/L. The activation of ERK was inhibited obviously by PTEN. Conclusion EGF can activate ERKs in Ishikawa cells. Lipid phosphatase PTEN can suppress activation of ERKs induced by EGF.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2002年第4期369-372,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金资助
No.3 0 0 0 0 178
