肺癌细胞自分泌VEGF对mCRPs的调控及其机制  

Autocrined VEGF regulates mCRPs expressions in lung cancer cells and the related mechanisms

作　　者：赵勇[1] 张玲[1] 李翠玲[1] 阴海鹏[1] 顾洪涛[1] 温培娥[1] 

机构地区：[1]山东省医学科学院基础医学研究所,山东省肿瘤免疫与中药免疫重点实验室,山东省现代医用药物与技术重点实验室,山东济南250062

出　　处：《中国肿瘤生物治疗杂志》2010年第3期308-312,共5页Chinese Journal of Cancer Biotherapy

基　　金：山东省自然科学基金资助项目(No.Y2007C154)~~

摘　　要：目的:研究人非小细胞肺癌A549细胞自分泌VEGF对膜结合补体调节蛋白(membrane-bounal complement regulatory proteins,mCRPs)的调控及其机制。方法:RT-PCR法检测CD46、CD55、CD59、VEGF及其受体(KDR和FLT-1)和IL-8及其受体(CXCR1 CXCR2)mRNA在人非小细胞肺癌A549细胞的表达。MTT法检测抗VEGF抗体和抗IL-8抗体对A549细胞增殖的影响,流式细胞术检测抗VEGF抗体和抗IL-8抗体对A549细胞mCRPs表达水平的影响,Western blotting检测抗VEGF抗体对转录因子KLF2和磷酸化NF-κB p65蛋白表达的影响。结果:A549细胞表达膜结合型CD46、CD55和CD59 mRNA,亦表达VEGF及其受体(KDR和FLT-1)和IL-8及其受体(CXCR1和CXCR2)mRNA。抗VEGF抗体明显抑制A549细胞的增殖(P<0.05)。终质量浓度0.1μg/ml抗VEGF抗体封闭72 h,CD55和CD59 mRNA表达下降,膜结合的CD55和CD59蛋白分子表达降低(均P<0.05);胞质和核KLF2蛋白相对定量值分别从0.63和0.88下降至0.42和0.66,胞质和胞核磷酸化NF-κB p65蛋白相对定量值分别从0.44和0.28下降至0.37和0.19。结论:A549细胞可能通过自分泌VEGF增加NF-κB p65和KLF2转录因子水平,从而上调CD55和CD59的表达。Objective：To explore the regulatory effect of autocrined VEGF on membrane-bound complement regulatory proteins（mCRPs） expression in lung cancer A549 cells and the involved mechanisms.Methods： mRNA expressions of CD46,CD55,CD59,VEGF and their receptors（KDR and FLT-1）,and IL-8 and its receptors（CXCR1 CXCR2） in A549 cells were detected by RT-PCR.The effects of anti-VEGF antibody and anti-IL-8 antibody on the proliferation and mCRPs expression in A549 cells were examined by MTT assay and flow cytometry,respectively.The effects of anti-VEGF antibody on the expression of transcription factor KLF2 and phospho-NF-κB p65 were examined by Western blotting analysis.Results： In addition to membrane CD46,CD55 and CD59 mRNA,both VEGF,IL-8 and their receptors（KDR,FLT-1;CXCR1,CXCR2） mRNA were expressed in A549 cells.Anti-VEGF antibody significantly inhibited the proliferation of A549 cells（P〈0.05）.CD55 and CD59 mRNA and protein levels in A549 cells were decreased after treatment with 0.1 μg/ml anti-VEGF antibody for 72 h（P〈0.05）.The relative values of KLF2 in cytoplasm and nuclear decreased from 0.63 and 0.88 to 0.42 and 0.66 after treatment with anti-VEGF antibody,while those of phospho-NF-κB p65 decreased from 0.44 and 0.28 to 0.32 and 0.19.Conclusion： VEGF may enhance CD55 and CD59 expressions in A549 cells through up-regulating expressions of NF-κB P65 and KLF2 transcription factors in an autocrine manner.

关 键 词：膜结合型补体调节蛋白(mCRPs) 血管内皮细胞生长因子 白细胞介素8 KLF2 NF-ΚB 肺肿瘤 

分 类 号：R730.2[医药卫生—肿瘤] R734.2[医药卫生—临床医学]