前列腺癌细胞核内雄激素受体靶基因的筛选

Screening of androgen receptor target genes in the nucleus of prostate cancer

目的筛选雄激素依赖性前列腺癌(PC)和雄激素非依赖性PC细胞核内的雄激素受体靶基因。方法取雄激素依赖性PC细胞株LNCaP和雄激素非依赖性PC细胞株LNCaP-AI进行培养,使用染色质免疫共沉淀技术在全基因组水平上筛选LNCaP、LNCaP-AI细胞中的雄激素受体结合位点,联合高通量测序技术寻找雄激素受体靶基因。采用RT-PCR法检测并比较LNCaP、LNCaP-AI细胞中雄激素受体差异靶基因表达水平。结果LNCaP、LNCaP-AI细胞中分别发现9021、4949个基因组测序数据富集区域,其中4143、2380个落在雄激素受体结合位点,将这些位点定位到基因组后分别得到2796、1854个雄激素受体相关基因(相同的基因有789个)。多数靶基因集中在肌动蛋白细胞骨架的调节、紧密连接、血管平滑肌收缩信号通路;LNCaP与LNCaP-AI细胞的差异靶细胞通路为黏着斑通路。从LNCaP与LNCaP-AI细胞测序得到的雄激素受体差异靶基因中筛选出富集程度较高的雄激素受体差异靶基因2个,即LIFR、PSMA6。LNCaP-AI细胞中LIFR、PSMA6 mRNA表达水平均明显高于LNCaP细胞(均P<0.05)。结论LIFR、PSMA6为PC细胞核内雄激素受体靶基因。

Objective To screen androgen receptor target genes in nuclear of androgen dependent prostate cancer(PC)and androgen independent PC.Methods Androgen dependent PC cell line LNCaP and androgen independent PC cell line LNCaP-AI were cultured,and the androgen receptor binding sites in LNCaP and LNCaP-AI cells were screened at the whole genome level by chromatin immunoprecipitation technique,combined with high-throughput sequencing technology to find androgen receptor target genes.RT-PCR was used to detect and compare the expression levels of androgen receptor differential target genes in LNCaP and LNCaP-AI cells.Results Nine thousand and twenty-one and four thousand nine hundred and forty-nine genome sequencing data rich regions were found in LNCaP and LNCaP-AI cells respectively,of which 4143 and 2380 were located at androgen receptor binding sites.After these sites were located in the genome,2796 and 1854 androgen receptor related genes were obtained respectively(789 of the same genes).Most of the target genes are concentrated in the regulation of actin cytoskeleton,tight junction and vascular smooth muscle contraction signal pathway;the different target cell pathway between LNCaP and LNCaP-AI cells is Adhesion patch pathway.From the androgen receptor differential target genes sequenced by LNCaP and LNCaP-AI cells,two androgen receptor differential target genes with high enrichment were screened,namely LIFR and PSMA6.The expression levels of LIFR and PSMA6 mRNA in LNCaP AI cells were significantly higher than those in LNCaP cells(all P<0.05).Conclusion LIFR and PSMA6 are androgen receptor target genes in PC nuclear.