摘要
目的探讨类表皮生长因子域8（EGFL8）基因沉默对非小细胞肺癌裸鼠模型瘤内血管生成的影响。方法构建EGFL8短发夹状RNA表达质粒并转染NCI-H358细胞（psh EGFL8组）,以空质粒转染为对照组,观察两组皮下种植瘤生长曲线及体积;免疫组织化学法检测抗CD34、血管内皮生长因子（VEGF）、血小板反应蛋白（TSP1）表达,RT-PCR检测MMP-2和TIMP2表达。结果 psh EGFL8组种植瘤体积为（1.75±0.75）cm3,微血管密度（MVD）为（20.14±5.18）,分级为1~2级;对照组种植瘤体积为（4.16±1.05）cm3,MVD为（40.14±5.18）,分级为3~4级;两组种植瘤体积、MVD和分级的差异有显著性,P值均〈0.05;EGFL8组TSP1蛋白阳性表达,而VEGF蛋白为弱阳性或阴性表达,MMP-2 mRNA表达下调,而TIMP2 mRNA表达上调,与对照组比较差异有显著性,P值均〈0.01。结论 EGFL8基因沉默可降低非小细胞癌癌种植瘤血管生成,与调节MMP-2/TIMP2表达,影响VEGF/TSP1平衡有关。
【Objective】To investigate the effect of epidermal growth factor-like domain 8（EGFL8） gene silencing on angiogenesis of non-small cell lung cancer in nude mice and its mechanism. 【Methods】A short hairpin RNA（shRNA） vector targeting at EGFL8 genewas constructed and transfected into NCI-H358 cell（pshEGFL8group）, and cells transfected with blank plasmids served as control group. Positive clones were selected and transplanted in nude mice to produce tumor animal mode. Growth curves and volumes of tumors were observed. After 8weeks, EGFL8 mRNA and protein in transplanted tumor tissues were detected and graded, and CD34, VEGF and TSP1 tested with immunochemistry method,and MMP-2 and TIMP2 mRNA detected with RT-PCR. 【Results】In pshEGFL8 group, the tumor volume was（1.75±0.75） cm3, and MVD was（20.14±5.18）, while in control group, tumor volume was（4.16±1.05） cm3, MVD was（40.14±5.18）. There were significant differences in two groups（P〈0.05）. TSP1 protein expressed positively, and VEGF protein presented weakly or negatively in pshEGFL8 group. The expression of MMP-2 mRNA decreased, while TIMP2 mRNA increased in pshEGFL8 group, and there were significant differences in two groups（P〈0.05）.【Conclusions】Silencing of EGFL8 gene can decrease angiogenesis of non-small cell lung cancer in nude mice, which might relate to regulation of expression of MMP-2 and TIMP2 and influence on balancing of TSP1/VEGF.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2014年第24期21-25,共5页
China Journal of Modern Medicine
基金
湖南省社会发展科技支撑计划重点项目(No:2013SK3198)
关键词
非小细胞肺癌
表皮生长因子
血管
non-small cell lung cancer epidermal growth factor blood vessels
