本文選題：宮頸癌　切入點：EFEMP　出處：《中華腫瘤防治雜志》2015年21期

【摘要】：目的前期研究已證實EFEMP1能促進宮頸癌微血管增生,本研究旨在進一步探討EFEMP1促進宮頸癌微血管生成的分子機制。方法利用Pierce CO-IP試劑盒檢測宮頸癌細胞中EFEMP1蛋白和表皮生長因子受體(epidermal growth factor receptor,EGFR)的相互作用,蛋白質(zhì)印跡法檢測蛋白表達。利用MTT實驗檢測內(nèi)皮細胞增殖活力,Transwell小室法檢測內(nèi)皮細胞遷移能力,Matrigel膠管腔形成能力實驗檢測內(nèi)皮管腔形成能力。構建慢病毒干擾顆粒干擾宮頸癌細胞Jagged1表達。利用免疫組化MaxVisonTM法檢測組織蛋白表達,內(nèi)皮CD34組化標記結合Weinner計數(shù)法檢測組織中微血管密度(microvsacular density,MVD)。皮下接種Hela-EFEMP1+細胞構建高表達EFEMP1的裸鼠荷瘤模型。結果 p-EGFR和p-MAPK在EFEMP1處理組Hela細胞的表達水平(2.35±0.41和2.56±0.44)分別高于對照組(0.20±0.001,P=0.001;0.64±0.020,P=0.001)和PD153035與EFEMP1聯(lián)合處理組(1.25±0.33,P=0.004;1.46±0.24,P=0.001)。CO-IP結果顯示,Hela細胞中EFEMP1與EGFR相互作用。處理組內(nèi)皮細胞24h活力(1.78±0.048)、遷移能力(71.7±4.91)和管腔形成能力(19.7±1.75)分別高于對照組(1.38±0.046,P=0.001;30±3.46,P=0.002 3;8.7±1.84,P=0.001 8)和聯(lián)合處理組(1.51±0.072,P=0.004;37.6±4.98,P=0.008 3;12.6±1.48,P=0.009 3)。處理組癌細胞中p-MAPK、血管內(nèi)皮生長因子(vascular endothelia growth factor,VEGF)和Jagged1的表達水平(0.96±0.05,1.25±0.031,0.82±0.036)分別高于對照組(0.16±0.006,P=0.028;0.54±0.047,P=0.013;0.42±0.017,P=0.026)和U0126與EFEMP1聯(lián)合處理組(0.56±0.022,P=0.042;0.29±0.016,P=0.008;0.38±0.037,P=0.024)。EFEMP1處理Hela-Jagged1siRNA細胞后上清液作用下的內(nèi)皮管腔形成能力(14±1.58,P=0.006)和γ-SI與處理組正常癌細胞上清液聯(lián)合作用的內(nèi)皮細胞管腔形成能力(12.6±1.33,P=0.008),分別低于處理組癌細胞上清液作用下的內(nèi)皮細胞(19.7±0.89)。宮頸癌組織中,EFEMP1表達分別與p-MAPK和Jagged1表達正相關,p-MAPK表達與Jagged1表達正相關,且p-MAPK和Jagged1的表達分別與宮頸癌MVD呈正相關。動物實驗結果顯示,LV-JAG1-RNAi慢病毒處理組腫瘤平均體積和質(zhì)量〔(0.160±0.007)cm3;(0.43±0.043)g〕分別明顯小于對照組〔(0.25±0.015)cm3,P=0.001;(0.65±0.027)g,P=0.002〕。此外,處理組平均MVD(5.0±1.23)也低于對照組(8.7±1.15,P=0.014)。結論 EFEMP1促進宮頸癌微血管生成的分子機制是通過與EGFR相互作用,激活MAPK-VEGF/Jagged1信號通路,上調(diào)宮頸癌VEGF和Jagged1的表達而旁分泌促進內(nèi)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.澶勭悊緇勭檶緇嗚優(yōu)涓璸-MAPK,琛，

本文編號：1730747