西安交通大学学报(医学版)
西安交通大學學報(醫學版)
서안교통대학학보(의학판)
Journal of Xi'an Jiaotong University (Medical Sciences)
2015年
6期
739-742
,共4页
血管生成拟态%缺氧%胶质瘤%EGFR/AKT/PI3K 信号通路
血管生成擬態%缺氧%膠質瘤%EGFR/AKT/PI3K 信號通路
혈관생성의태%결양%효질류%EGFR/AKT/PI3K 신호통로
vasculogenic mimicry%hypoxia%glioma%EGFR/AKT/PI3K signaling pathway
目的:探讨表皮生长因子受体(epithelial growth factor receptor,EGFR)对血管生成拟态的促进作用。方法应用 CD34-PAS 双重染色法检测不同级别胶质瘤中血管生成拟态的存在情况,应用免疫组织化学法检测不同级别胶质瘤中 EGFR 蛋白的表达,应用化学缺氧法诱导人脑星形细胞瘤 U87细胞在三维培养体系中形成血管生成拟态, Transwell法检测细胞的迁移及侵袭能力,甲基噻唑基四唑法(methyl thiazolyl tetrazolium,MTT)检测细胞的增殖能力,蛋白质印迹法(Western blot)检测 EGFR、丝氨酸苏氨酸蛋白激酶 AKT 和磷脂酰肌醇3激酶(phosphatidylinositol 3 kinase,PI3K)蛋白的表达,探讨 EGFR/AKT/PI3K 通路对血管生成拟态的诱导作用。结果血管拟态阳性的患者EGFR 阳性率为94.4%(17/18),血管拟态阴性的患者 EGFR 阳性率为71.8%(56/78),两者差异具有统计学意义(P <0.05)。U87细胞三维培养后,缺氧组平均每个视野网状结构数量、迁移能力,侵袭能力和增殖能力均显著高于常氧组(P <0.01)。Western blot 显示缺氧组中 EGFR、AKT、PI3K 蛋白的表达显著高于常氧组。结论缺氧可诱导EGFR/AKT/PI3K 信号通路活化,进而诱导血管生成拟态的形成,EGFR 在血管生成拟态的形成过程中起着十分重要的作用。
目的:探討錶皮生長因子受體(epithelial growth factor receptor,EGFR)對血管生成擬態的促進作用。方法應用 CD34-PAS 雙重染色法檢測不同級彆膠質瘤中血管生成擬態的存在情況,應用免疫組織化學法檢測不同級彆膠質瘤中 EGFR 蛋白的錶達,應用化學缺氧法誘導人腦星形細胞瘤 U87細胞在三維培養體繫中形成血管生成擬態, Transwell法檢測細胞的遷移及侵襲能力,甲基噻唑基四唑法(methyl thiazolyl tetrazolium,MTT)檢測細胞的增殖能力,蛋白質印跡法(Western blot)檢測 EGFR、絲氨痠囌氨痠蛋白激酶 AKT 和燐脂酰肌醇3激酶(phosphatidylinositol 3 kinase,PI3K)蛋白的錶達,探討 EGFR/AKT/PI3K 通路對血管生成擬態的誘導作用。結果血管擬態暘性的患者EGFR 暘性率為94.4%(17/18),血管擬態陰性的患者 EGFR 暘性率為71.8%(56/78),兩者差異具有統計學意義(P <0.05)。U87細胞三維培養後,缺氧組平均每箇視野網狀結構數量、遷移能力,侵襲能力和增殖能力均顯著高于常氧組(P <0.01)。Western blot 顯示缺氧組中 EGFR、AKT、PI3K 蛋白的錶達顯著高于常氧組。結論缺氧可誘導EGFR/AKT/PI3K 信號通路活化,進而誘導血管生成擬態的形成,EGFR 在血管生成擬態的形成過程中起著十分重要的作用。
목적:탐토표피생장인자수체(epithelial growth factor receptor,EGFR)대혈관생성의태적촉진작용。방법응용 CD34-PAS 쌍중염색법검측불동급별효질류중혈관생성의태적존재정황,응용면역조직화학법검측불동급별효질류중 EGFR 단백적표체,응용화학결양법유도인뇌성형세포류 U87세포재삼유배양체계중형성혈관생성의태, Transwell법검측세포적천이급침습능력,갑기새서기사서법(methyl thiazolyl tetrazolium,MTT)검측세포적증식능력,단백질인적법(Western blot)검측 EGFR、사안산소안산단백격매 AKT 화린지선기순3격매(phosphatidylinositol 3 kinase,PI3K)단백적표체,탐토 EGFR/AKT/PI3K 통로대혈관생성의태적유도작용。결과혈관의태양성적환자EGFR 양성솔위94.4%(17/18),혈관의태음성적환자 EGFR 양성솔위71.8%(56/78),량자차이구유통계학의의(P <0.05)。U87세포삼유배양후,결양조평균매개시야망상결구수량、천이능력,침습능력화증식능력균현저고우상양조(P <0.01)。Western blot 현시결양조중 EGFR、AKT、PI3K 단백적표체현저고우상양조。결론결양가유도EGFR/AKT/PI3K 신호통로활화,진이유도혈관생성의태적형성,EGFR 재혈관생성의태적형성과정중기착십분중요적작용。
Objective To investigate the effect and mechanism of EGFR in inducing vasculogenic mimicry (VM)formation.Methods Immunohistochemistry was used to detect the expression of EGFR protein and CD34-PAS double staining was used to detect the VM in gliomas paraffin blocks.The migration and VM formation of U87 cell in hypoxic and normoxic groups were detected by transwell and three-dimensional culture.Then we used Western blot to detect the expressions of EGFR,AKT and PI3K protein.Results The positive rate of EGFR expression in VM positive patients was 94.4%,which was significantly higher than that in VM negative patients (71.8%).In hypoxic group,the abilities of migration and VM formation were significantly higher those in normoxic group.The expressions of EGFR,AKT and PI3K were higher in hypoxic group.Conclusion VM formation can be induced by the activation of EGFR/AKT/PI3K signaling pathway.EGFR is very important for VM formation.