【摘要】 目的:探讨在ErbB2非过表达乳腺癌细胞MDA-MB-231中,神经调节因子(neuregulins,NRGs)对ErbB2受体信号转导活化和侵袭转移能力的作用。方法:免疫细胞化学法和Western blot检测MDA-MB-231细胞中neuregulin(NRG)的表达o应用ErbB2受体功能抑制剂AG825处理MDA-MB-231,MTT法检测细胞的增殖抑制作用:Transwell小室进行人工重组基底膜侵袭和迁移实验;明胶酶谱实验检测MMP-2、MMP—9的活性。Western blot检测MMP-2、HIF-1α蛋白表达。结果:神经调节因子在乳腺癌细胞MDA-MB—231呈显著表达,Western blot实验可见分子量44KD的NRG抗体阳性反应条带。应用ErbB2受体功能抑制剂AG825后,MDA-MB-231细胞生长受到抑制,作用呈时效量效依赖关系(P<0.01);细胞的侵袭、迁移能力均明显下降(P<0.05),MMP-2、MMP—9的活性降低(P<0.05);MMP-2、HIF-1α蛋白表达减少(P<0.05)。结论:ErbB2非过表达乳腺癌细胞MDA-MB-231存在神经调节因子分泌,可能通过神经调节因子自分泌或旁分泌作用方式使ErbB2受体信号转导处于激活状态,从而影响细胞侵袭转移能力。更多还原

【Abstract】 Objective:To explore the effect of neuregulins on ErbB2 receptor signal transduction pathway activation,and invasion and metastasis of none-overexpression ErbB2 breast cancer cell MDA-MB-231.Methods: The expressions of neuregulin were detected by immunocytochemistry and Western blot.MDA-MB-231 cells were treated with ErbB2 kinase inhibitor AG825.Proliferation were measured with MTT assay.Invasion and metastasis of MDA-MB-231 cells were evaluated with transwell chamber.The expression of MMP-2,MMP-9 enzymatic activity was detected by gelatin zymography.The expressions of MMP-2 and HIF-1α were detected by Western blot.Results: MDA-MB-231 cells expressed a relative higher level of neuregulin.In the western blot,the positive reaction band has been found in the location where the molecular weight of the NRG is 44KD.When MDA-MB-231 cells were treated with AG825,the proliferation was inhibited on time-dose-depending manner(P<0.01),invasion and metastasis were also depressed(P<0.05).The expression of MMP-2 and MMP9 enzymatic activity was lower(P<0.05).The expression level of MMP-2 and HIF-1 α were decreased(P<0.05).Conclusion: Our study indicates that neuregulins were synthesized in MDA-MB-231 cells as transmembrane proteins,neuregulins could active ErbB2 receptor signal transduction pathway by autocrine or paracrine action,and induced invasion and metastasis of MDA-MB-231 cells.更多还原