Impact of splicing factors (ESRP1 and KHDRBS3) on prostate cancer biology. Impact of splicing factors (ESRP1 and KHDRBS3) on prostate cancer biology

Dysregulation of mRNA alternative splicing (AS) has been implicated in development and progression of hematological malignancies. How the global AS dysregulation contributes to the development and progression of solid tumors remains generally unclear. Recently, we show that many splicing factors (such as ESRP1 and KHDRBS3) are overexpressed in human primary prostate cancer (PCa) versus normal tissues. To further determine the biological impact of splicing factors on PCa aggressiveness, we treated AR- PC3 cells with siRNAs against either ESRP1 or KHDRBS3 and both. RNA-seq analysis was done in triplicates of rRNA-depleted total RNAs isolated from PC3 cells 48-72hr after siRNA infection. rMATS and DEseq2 were used to reveal the effect of knocking down ESRP1 and/or KHDRBS3 individually or in combination on mRNA splicing landscape and gene expression change. Overall design: Quantification of changes in RNA splicing and gene expression in prostate cancer cells treated with siRNAs to knockdown either ESRP1 or KHDRBS3 individually and in combination.

mRNA可变剪接（alternative splicing，AS）失调已被证实与血液系统恶性肿瘤的发生发展密切相关。然而，全局AS失调如何推动实体瘤的发生发展，目前仍未得到清晰阐明。近期本研究团队发现，相较于正常组织，多种剪接因子（splicing factor）在人类原发性前列腺癌（prostate cancer，PCa）组织中呈高表达，其中包括ESRP1与KHDRBS3。为进一步明确剪接因子对PCa侵袭性的生物学影响，本研究对雄激素受体阴性（AR-）PC3细胞，分别采用靶向ESRP1、KHDRBS3的小干扰RNA（small interfering RNA，siRNA）及二者联合的siRNA进行处理。本研究对转染siRNA后48至72小时的PC3细胞中提取的去除核糖体RNA的总RNA，设置三次生物学重复进行RNA测序（RNA-seq）分析。采用rMATS与DEseq2两款分析工具，分别解析单独或联合敲低ESRP1、KHDRBS3对mRNA剪接图谱及基因表达变化的影响。实验整体设计：对分别或联合使用靶向ESRP1、KHDRBS3的siRNA处理的前列腺癌细胞中RNA剪接与基因表达的变化进行定量分析。