冬凌草甲素下调EGFR/ERK/MMP-12和CIP2A/Akt信号通路抑制gefitinib耐受的非小细胞肺癌的转移  

Oridonin Inhibits Gefitinib-resistant Non-small-cell Lung Cancer Cells Metastasis by Down-regulating EGFR/ERK/MMP-12 and CIP2A/Akt Signal Pathways

作　　者：肖香玲 何中维 黄秋月[1] 季娟丽 张云飞[1] 刘莹[1] 

机构地区：[1]湖北医药学院基础医学院,湖北十堰442000 [2]温州医科大学附属第一医院,浙江温州325035

出　　处：《湖北医药学院学报》2016年第6期536-542,共7页Journal of Hubei University of Medicine

基　　金：2014年度湖北医药学院优秀中青年科技创新团队资助计划项目(2014 CXX05);湖北医药学院2015年度研究生启动基金资助计划项目(2015QDJZR16);2016年大学生创新创业训练计划项目(201610929001)

摘　　要：目的:研究天然化合物冬凌草甲素(oridonin,Ori)抑制吉非替尼(gefitinib)药物耐受的非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞转移的作用及其机制。方法:用不同浓度的Ori作用于肺癌H1975细胞,划痕修复实验、Transwell法检测Ori对细胞迁移和侵袭能力的影响;Western blot检测Ori对细胞EGFR及下游信号通路,Akt及相关因子蛋白表达水平的影响;明胶酶谱检测金属基质蛋白蛋白酶12(matrix metalloproteinase-12,MMP-12)活性;荧光素酶报告系统检测Ori对AP-1转录活性的影响;Ch IP实验检测Ori对转录因子c-Fos和MMP-12结合的影响。结果:Ori能显著抑制细胞侵袭、迁移和粘附能力。Ori浓度依赖性抑制EGFR、ERK和Akt磷酸化,抑制MMP-12和癌蛋白(cancerous inhibitor of protein phosphatase 2A,CIP2A)表达,而上调其底物蛋白磷酸酶2A(protein phosphatase 2A,PP2A)的表达及活性。Ori抑制转录因子AP-1的转录活性及c-Fos和MMP-12启动子结合。结论:Ori可以通过抑制EGFR/ERK/MMP-12和CIP2A/PP2A/Akt信号通路来抑制gefitinib耐药的NSCLC细胞的侵袭和迁移。Objective To explore the effects and mechanisms of natural compound oridonin （Ori） inhibit the cells metastasis of gefifinib-resistant non-small cell lung cancer （NSCLC）. Mothods Lung cancer H1975 cells were treated with different concentrations Ori, then the abilities of migration and invasion were detected by wotmd healing assay and transwell assay; Western blot was used to detect the influence of Ori on EGFR and its downstream signal pathway, Akt and correlated protein expression; gelatin zymography was used to exam the gelatinolytie activity of matrix metalloproteinase-12 （MMP-12） ; lueiferase reporter assay was used to detect the transcription aetivity of AP- 1. Chromatin immunopreeipitation （CHIP） assay was used to test the DNA binding of e-Fos （component of AP-1） to MMP-12 promoter. 1：to81.1118 The cells invasion, migration and adhesion were obviously inhibited by Off ; the phoSphoryiation of EGFR, ERK and Akt, expressions of MMP- 12 and CIP2A were inhibited by Ori in a dose-dependent manner; but the expression and ability of PP2A were-down-regulated by Off. The transcription activity of AP-1, DNA binding of c-Fos to MMP-12 promoter were both inhibited by Off. Conclusion Ori could inhibit the invasion and migration of geftinib-resistant NSCLC cells by down-regulating EGFR/ERK/MMP-12 and CIP2A/PP2A/Akt signal pathways.

关 键 词：冬凌草甲素 gefitinib耐药 肺癌 EGFR CIP2A 

分 类 号：R734.2[医药卫生—肿瘤]