非小细胞肺癌中PAX6表达及其对肿瘤细胞增殖与侵袭作用机制研究  

Expression and interaction mechanism of PAX6 in human non-small cell lung cancer

作　　者：熊安洲[1] 陈菁[1] 曹艳[1] 胡小萍[1] 

机构地区：[1]武汉市第一医院呼吸科,湖北武汉430022

出　　处：《中国癌症杂志》2014年第8期604-609,共6页China Oncology

基　　金：武汉市卫计委科研课题(No:WX14B03)

摘　　要：背景与目的:转录因子PAX6主要表达于胚胎期,不同肿瘤中PAX6呈现高表达,通过不同的信号通路发挥肿瘤抑制或促进作用。检测PAX6在非小细胞肺癌(non-small cell lung cancer,NSCLC)中的表达,通过RNAi下调其表达,研究PAX6对肿瘤细胞侵袭和增殖力以及cyclin E、p38表达水平的影响。方法:应用蛋白印记检测86例NSCLC肿瘤组织及癌旁配对组织以及2例细胞系中PAX6的表达情况。应用PAX6特异性siRNA序列,转染NSCLC细胞系A549,MTT法、Transwell小室、划痕实验检测转染前后细胞增殖、侵袭和迁移能力的对比。蛋白质印记法(Western blot)检测转染前后cyclin E及p38表达情况。结果:对比癌旁组织及正常人支气管上皮细胞16HBE,PAX6在NSCLC组织及A549细胞系中显著高表达。选取高效siRNA序列转染细胞系后,PAX6表达的下调抑制了肿瘤细胞的增殖及集落形成,G1期细胞比例增加,细胞侵袭及迁移力均下降。PAX6基因敲低细胞cyclin E及p38活性受到抑制,表达下调。结论:PAX6通过调控MAPK通路及cyclin E的表达加速肿瘤细胞的增殖、侵袭及迁移,是潜在的NSCLC诊疗靶点。Background and purpose：The transcription factor PAX6 is primarily expressed in embryos. PAX6 is also expressed in several tumors and plays oncogenic or tumor suppressor role. This study aimed to investigate the expression of PAX 6 in non-small cell lung cancer （NSCLC） tumor samples and cell lines; and evaluate the effects of PAX6 on the proliferation and invasion of tumor cells and the expression level of cyclin E and p38.Methods：Western blot was carried out to detect the PAX6 protein level in 86 NSCLC tumor tissues, paired adjacent normal tissues and 2 cell lines. PAX6 siRNA was transfected into human lung cancer A549 cell line. Anchorage-independent growth and invasiveness of tumor cells were measured by MTT and transwell cell invasion assay, respectively. Cyclin E and p38 protein level before and after transfection were detected by western blot.Results：Comparison with tumor adjacent tissues and normal human bronchial epithelial cells 16HBE, PAX6 in NSCLC tissues and A549 cell lines was signiifcantly higher expression. After transfection with efifcient sequence of PAX6 siRNA for A549 cell lines, the down expression of PAX6 inhibits tumor cell proliferation, the proportion of cells in G1 phase increased, the cell invasion and migration decreased remarkably. Cyclin E and p38 activity was inhibited inPAX6 knockdown cells.Conclusion：PAX6 accelerate cell cycle progression by activating cyclin E and p38. PAX6 is a potential target for diagnosis and therapy.

关 键 词：非小细胞肺癌 PAX6 SIRNA P38 CYCLIN E 

分 类 号：R734.2[医药卫生—肿瘤]