PTEN loss and expression of the TMPRSS2/ERG fusion gene transform prostatic epithelial cells via enhanced FGF signaling. Homo sapiens

To determine if ERG expression and PTEN loss were sufficient to transform prostate epithelial cells we constructed cell lines with expression of the TMPRSS2/ERG fusion gene (TE), stable knockdown of PTEN with shRNA (PTEN KD) or both alterations (PTEN KD/TE) using the PNT1A cell line. To determine what gene expression changes are associated the phenotypic changes between the cell line experimental groups, we carried expression microarray studies using Agilent 60K expression microarrays. RNAs from all four cell lines were analyzed in duplicate and probes with ≥ 1.4-fold or ≤0.7-fold relative to control cells identified. Our results demonstrated that PTEN loss and expression of the TMPRSS2/ERG fusion gene transform prostatic epithelial cells via enhanced FGF signaling. Overall design: Microarray analyses of four cell lines (PA, PA-TE, PA-PTEN KD and PA-TE+PTEN KD).

为明确ERG表达与PTEN缺失是否足以转化前列腺上皮细胞，我们以PNT1A细胞系为基础构建了三类工程细胞株：仅过表达TMPRSS2/ERG融合基因（TE）、通过短发卡RNA（shRNA）稳定敲低PTEN（PTEN KD），以及同时携带两种遗传改变的细胞株（PTEN KD/TE）。为探究与各细胞株实验组间表型变化相关的基因表达变化，我们采用安捷伦(Agilent)60K表达芯片开展了表达微阵列分析。我们对全部四组细胞株的RNA样本进行了重复检测，并筛选出相对对照组表达量≥1.4倍或≤0.7倍的探针。本研究结果表明，PTEN缺失与TMPRSS2/ERG融合基因的表达可通过增强成纤维细胞生长因子(FGF, Fibroblast Growth Factor)信号通路实现前列腺上皮细胞的恶性转化。整体实验设计：对四组细胞株（PA、PA-TE、PA-PTEN KD及PA-TE+PTEN KD）进行表达微阵列分析。