雄激素非依赖性前列腺癌细胞雄激素受体靶基因的筛选与初步鉴定  

The Screening and Identifi cation of Androgen Receptor Target Genes in Androgen-independent Prostate Cancer Cell

作　　者：徐思琪[1,2] 廖昭平 刘春华[3,2] 程玥[2] 季丽丽[1,2] 段秀枝[2] 陈玉华[2] 陶志华[1,2] 

机构地区：[1]温州医科大学附属第一医院实验诊断中心,温州325000 [2]浙江大学医学院附属第二医院临床检验中心,杭州310009 [3]温州医科大学检验医学院生命科学学院,温州325000

出　　处：《中国细胞生物学学报》2014年第7期941-948,共8页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81271917)资助的课题~~

摘　　要：采用染色质免疫共沉淀技术在全基因组水平筛选雄激素非依赖性前列腺癌细胞LNCaP-AI的雄激素受体结合位点,行高通量测序及生物信息学分析共得到2 876个peak(pvalue<1×10–5),peak平均长度为673 bp;将peak序列定位到Hg19基因组,共有1 865个靶基因,其中fold enrichment≥10的基因有425个。对peak相关基因进行GO分析发现,与细胞、细胞组分、细胞过程、结合、细胞器相关的基因位列前五位;对peak相关基因进行通路分析发现,与黏着斑、代谢通路、癌症中的转录错误调控、嘌呤代谢等信号通路相关的基因占大多数。筛选出7个候选AR靶基因,采用Real-time qPCR技术分析它们在LNCaP-AI细胞和雄激素依赖性前列腺癌细胞LNCaP中对DTH刺激的反应性,发现DHT刺激可改变7个候选AR靶基因在LNCaP-AI细胞中的表达,为进一步研究雄激素依赖性前列腺癌向非依赖性前列腺癌发展的过程中雄激素受体及其调控的下游靶基因功能起着至关重要的作用。Chromatin immunoprecipitation assay was performed to screen the androgen receptor binding sites in androgen-independent prostate cancer cell LNCaP-AI in the whole genomic. Using high-throughput sequencing and bioinformatic analysis, there were 2 876 peaks （p-value〈1× 10^-5）, and the average length of peaks was 673 bp; locating each peak in the Hg19 genome, 1 865 genes were founded. There were 425 genes which the fold enrichment was more than 10. It could be founded that the top five genes were associated with cell, cell part, cellular process, binding and organelle by GO analysis of peak related genes. Genes associated with focal adhesion, metabolic pathways, transcriptional misregulation in cancer and purine metabolism were the majority by pathway analysis of peak related genes. Seven candidated AR target genes were selected to analyze the reactivity to DHT stimulation in LNCaP-AI cell and LNCaP cell by Real-time qPCR. The result showed that DHT stimulation could change the expression of seven candidated target genes in LNCaP-AI cell. Our data play a vital role in the further study onandrogen receptor and it＇s regulated target genes in the process of androgen-dependent prostate cancer progress to androgen-independent prostate cancer.

关 键 词：前列腺癌 雄激素非依赖 染色质免疫共沉淀 雄激素受体 靶基因 

分 类 号：R737.25[医药卫生—肿瘤]