EFEMP1促进宫颈癌微血管生成分子机制研究

发布时间：2018-04-10 10:02

  本文选题：宫颈癌　切入点：EFEMP　出处：《中华肿瘤防治杂志》2015年21期

【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表达正相关,p-MAPK表达与Jagged1表达正相关,且p-MAPK和Jagged1的表达分别与宫颈癌MVD呈正相关。动物实验结果显示,LV-JAG1-RNAi慢病毒处理组肿瘤平均体积和质量〔(0.160±0.007)cm3;(0.43±0.043)g〕分别明显小于对照组〔(0.25±0.015)cm3,P=0.001;(0.65±0.027)g,P=0.002〕。此外,处理组平均MVD(5.0±1.23)也低于对照组(8.7±1.15,P=0.014)。结论 EFEMP1促进宫颈癌微血管生成的分子机制是通过与EGFR相互作用,激活MAPK-VEGF/Jagged1信号通路,上调宫颈癌VEGF和Jagged1的表达而旁分泌促进内皮血管生成。
[Abstract]:Objective to investigate the molecular mechanism of EFEMP1 promoting microangiogenesis in cervical cancer.Methods the interaction of EFEMP1 protein and epidermal growth factor receptor (EGFR) in cervical cancer cells was detected by Pierce CO-IP kit, and protein expression was detected by Western blot.MTT assay was used to detect endothelial cell proliferation activity and transwell chamber method was used to detect the migration ability of endothelial cells.Lentivirus interference particles were constructed to interfere with the expression of Jagged1 in cervical cancer cells.The expression of tissue protein was detected by immunohistochemical MaxVisonTM method, and the microvascular density (MVD) was detected by CD34 histochemistry and Weinner counting.Hela-EFEMP1 cells were inoculated subcutaneously to construct a tumor-bearing model of nude mice with high expression of EFEMP1.澶勭悊缁勭檶缁嗚優涓璸-MAPK,琛，

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