摘要
背景与目的:雌激素和胰岛素样生长因子1(insulinlikegrowthfactor1,IGF-1)在子宫内膜癌细胞中的信号转导机制尚不清楚。本文拟研究雌激素和IGF-1在子宫内膜癌细胞信号转导中激活细胞外信号调节激酶(extracellularsignal-regulatedkinase,ERK)的情况,探讨与细胞骨架蛋白同源的磷酸酶(phosphataseandtensinhomologue,PTEN)在雌激素和IGF-1诱导的ERKs活化中的作用。方法:构建野生型PTEN和突变型PTEN(G129E)的逆转录病毒载体,体外转染Ishikawa细胞,用G418筛选稳定表达的细胞系。Westernblot检测PTEN基因在Ishikawa细胞中的表达,并检测和观察17-β-雌二醇和IGF-1对细胞系Ishikawa-EGFP(对照)、Ishikawa-PTEN和Ishikawa-PTEN(G129E)ERKs活化的浓度效应和时相特点,以及17-β-雌二醇在瞬时转染pCXN2hERα和pCXN2hERβ的NIH3T3细胞中激活ERK的情况。结果:IGF-1可激活ERK,以ERK2为主。不同浓度IGF-1作用于Ishikawa-EGFP和Ishikawa-PTEN时,对ERK2活化无明显区别。40μg/L的IGF-1作用显示5min时ERK2活化最明显。PTEN在IGF-1作用Ishikawa30min、2h和24h时均能抑制ERKs活化,而在5min时没有明显作用。40μg/LIGF-1刺激Ishikawa-PTEN(G129E)5min,和Ishikawa-EGFP相比,无明显改变。17-β-雌二醇可激活ERK,以ERK2为主。不?
Background &Objectives:The machanism of signal transductio n of insulin-like growth factor(IGF-1)in endometrial carcinoma is still unknow.The objective of this paper was to study extracellular signal-regula ted kinase(ERK)activation in endometrial carcinom a cell line Ishikawa under the stimul ation of IGF-1,and to elucidate the r ole of suppressor encoprotein phosphatase and tensin homologue(PTEN)in activation of ERK.Methods:Retrovirus vector of PTEN and PTEN(G129E)was constructed.Ishikawa was transfected in vitro.Expressionstable c ell line was screened by G418.Western blot was applied to examine PTEN expression in Ishikawa cells after transfection.Optimal concen tration and time of IGF-1and17-β-estrodial which activated ERK in Is hikawa-PTEN and Ishikawa-PTEN(G129E)cells were detected.Western blot wa s applied to examine ERK activation un der the stimulation of17-β-estrodial in NIH3T3fibroblasts after transient transfection of pCXN2hERαand pCXN2hERβ.Results:IGF-1activated ERK cascades(mainly ERK 2 )in Ishikawa cells.There was no obvious difference in ERK activatio n among different doses of IGF-1(20,40,and 80μg /L),But the maximal activations of ERK 2 took place at 5min after stimulation with IGF-1.The activation of ERK 2 was inhibited obviously by PTEN at 30min,2h,and 24h.There was no obvious difference in ERK activation between Ish ikawa-PTEN(G129E)and Ishikawa-EGFP.17-β-estrodial activated ERK cascades(mainly ERK 2 )in Ishikawa cells.The activation of ERK was increased with increasing of concentration.The ma ximal activations of ERK 2 took place at 5min after stimulation with 17-β-estrodial.The activation of ERK 2 was inhibited obviously by PTEN,not by PTEN(G129E).17-β-estrodial activated ERKs cascades in NIH3T3fibroblasts after transient transduction of pCXN2h-ERα.Conclusions :17-β-estrodial and IGF-1activated ERK cascades in Ishikawa cells.Lipid phosphatase of PTEN had an inhibitory r ole in the activation of ERK under the stimulation of 17-β-estrodial and I
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2002年第7期724-730,共7页
Chinese Journal of Cancer
基金
国家自然科学基金项目(No.30000178)
