EFEMP1促进宫颈癌微血管生成分子机制研究  

Molecular mechanism through which EFEMP1 promotes the angiogenesis of cervical carcinoma

作　　者：宋恩霖[1] 张文辉[2] 熊秀娟[1] 况晓东[1] 俞薇薇[1] 

机构地区：[1]南昌大学基础医学院病理教研室,江西南昌330006 [2]中山大学附属第一医院病理科,广东广州510080

出　　处：《中华肿瘤防治杂志》2015年第21期1667-1674,共8页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然(青年)科学基金(81202054)

摘　　要：目的前期研究已证实EFEMP1能促进宫颈癌微血管增生,本研究旨在进一步探讨EFEMP1促进宫颈癌微血管生成的分子机制。方法利用Pierce CO-IP试剂盒检测宫颈癌细胞中EFEMP1蛋白和表皮生长因子受体(epidermal growth factor receptor,EGFR)的相互作用,蛋白质印迹法检测蛋白表达。利用MTT实验检测内皮细胞增殖活力,Transwell小室法检测内皮细胞迁移能力,Matrigel胶管腔形成能力实验检测内皮管腔形成能力。构建慢病毒干扰颗粒干扰宫颈癌细胞Jagged1表达。利用免疫组化MaxVisonTM法检测组织蛋白表达,内皮CD34组化标记结合Weinner计数法检测组织中微血管密度(microvsacular density,MVD)。皮下接种Hela-EFEMP1+细胞构建高表达EFEMP1的裸鼠荷瘤模型。结果 p-EGFR和p-MAPK在EFEMP1处理组Hela细胞的表达水平(2.35±0.41和2.56±0.44)分别高于对照组(0.20±0.001,P=0.001;0.64±0.020,P=0.001)和PD153035与EFEMP1联合处理组(1.25±0.33,P=0.004;1.46±0.24,P=0.001)。CO-IP结果显示,Hela细胞中EFEMP1与EGFR相互作用。处理组内皮细胞24h活力(1.78±0.048)、迁移能力(71.7±4.91)和管腔形成能力(19.7±1.75)分别高于对照组(1.38±0.046,P=0.001;30±3.46,P=0.002 3;8.7±1.84,P=0.001 8)和联合处理组(1.51±0.072,P=0.004;37.6±4.98,P=0.008 3;12.6±1.48,P=0.009 3)。处理组癌细胞中p-MAPK、血管内皮生长因子(vascular endothelia growth factor,VEGF)和Jagged1的表达水平(0.96±0.05,1.25±0.031,0.82±0.036)分别高于对照组(0.16±0.006,P=0.028;0.54±0.047,P=0.013;0.42±0.017,P=0.026)和U0126与EFEMP1联合处理组(0.56±0.022,P=0.042;0.29±0.016,P=0.008;0.38±0.037,P=0.024)。EFEMP1处理Hela-Jagged1siRNA细胞后上清液作用下的内皮管腔形成能力(14±1.58,P=0.006)和γ-SI与处理组正常癌细胞上清液联合作用的内皮细胞管腔形成能力(12.6±1.33,P=0.008),分别低于处理组癌细胞上清液作用下的内皮细胞(19.7±0.89)。宫颈癌组织中,EFEMP1表达分别与p-MAPK和Jagged1表达正�OBJECTIVE To investigate the mechanism of EFEMP1 in promoting the angiogenesis of cervical carci- noma. METHODS Pierce CO-IP was used to detect the interaction between EFEMP1 and EGFR in Hela cells. Western blotting was used to detect the protein expression level. The cell proliferative ability was measured by MTT, the endothelial cell migration capacity was assessed with Transwell Chamber method, and the tube formation capacity of endothelial cells was evaluated with Matrigel tube formation assay. A recombinant lentivirus was established to interfere the Jagged1 expression of cervical carcinoma cells. MaxVisonTM was adopted to assess the protein expression in tissues. The nude mice models of cervical carcinoma were established by implanting Hela ceils subcutaneously. RESULTS The expression levels of p-EGFR and p-MAPK （2.35±0.41, 2.56±0.44） in the Hela cells of EFEMP1 treatment group were significantly higher than the levels in the ceils of control group （0.02±0.001,P〈0. 001;0. 64±0. 020,P〈0. 001） and the group of combined treatment with PD153035 and EFEMPI（1.25±0.33,P=0. 004;1.46±0.24,P=0. 001）, respectively. The results of CO-IP indicated that EFEMP1 interacts with EGFR in Hela cells. The endothelial cells （ECs） of treatment group showed a significantly higher proliferative ability （1.78±0. 048）, migration capacity （71.7±4.91） and tube formation capacity （19.7±1.75） than the ECs of control group （1.38±0. 046, P〈0. 001;30-3.46, P=0. 002 3; 8.7± 1.84, P=0. 0018） and the ECs of combined treatment group （1.51±0. 072,P=0. 004;37.6±4.98,P=0. 008 3;12.6± 1.48,P=0. 009 3）, respectively. The expression levels of p-MAPK （0.96±0.05）, VEGF （1.25±0. 031） and Jagged1 （0.82±0. 036） in the Hela cells of treatment group were clearly higher than the levels in the cells of control group （0.16±0. 006,P=0. 028; 0.54±0. 047, P=0. 013; 0.42±0. 017, P=0. 026） and the ceils of the combined treatment group （0.56±0. 022, P=0. 042; 0.29±0. 016, P=0. 008; 0.3

关 键 词：宫颈癌 EFEMP1 微血管生成 分子机制 印迹法 蛋白质 

分 类 号：R737.33[医药卫生—肿瘤]