BACKGROUND Dysregulation of splicing variants (SVs) expression has recently emerged as a novel cancer hallmark. Although the generation of aberrant SVs (e.g. AR-v7/sst5TMD4/etc.) is associated to prostate-cancer (PCa) aggressiveness and/or castration-resistant PCa (CRPC) development, whether the molecular reason behind such phenomena might be linked to a dysregulation of the cellular machinery responsible for the splicing process [spliceosome-components (SCs) and splicing-factors (SFs)] has not been yet explored. METHODS Expression levels of 43 key SCs and SFs were measured in two cohorts of PCa-samples: 1) Clinically-localized formalin-fixed paraffin-embedded PCa-samples (n = 84), and 2) highly-aggressive freshly-obtained PCa-samples (n = 42). FINDINGS A profound dysregulation in the expression of multiple components of the splicing machinery (i.e. 7 SCs/19 SFs) were found in PCa compared to their non-tumor adjacent-regions. Notably, overexpression of SNRNP200, SRSF3 and SRRM1 (mRNA and/or protein) were associated with relevant clinical (e.g. Gleason score, T-Stage, metastasis, biochemical recurrence, etc.) and molecular (e.g. AR-v7 expression) parameters of aggressiveness in PCa-samples. Functional (cell-proliferation/migration) and mechanistic [gene-expression (qPCR) and protein-levels (western-blot)] assays were performed in normal prostate cells (PNT2) and PCa-cells (LNCaP/22Rv1/PC-3/DU145 cell-lines) in response to SNRNP200, SRSF3 and/or SRRM1 silencing (using specific siRNAs) revealed an overall decrease in proliferation/migration-rate in PCa-cells through the modulation of key oncogenic SVs expression levels (e.g. AR-v7/PKM2/XBP1s) and alteration of oncogenic signaling pathways (e.g. p-AKT/p-JNK). INTERPRETATION These results demonstrate that the spliceosome is drastically altered in PCa wherein SNRNP200, SRSF3 and SRRM1 could represent attractive novel diagnostic/prognostic and therapeutic targets for PCa and CRPC.

中文翻译：

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剪接机制的失调与前列腺癌的侵袭性直接相关。

背景技术近来，剪接变体（SVs）表达的失调已经成为一种新颖的癌症标志。尽管异常SV的产生（例如AR-v7 / sst5TMD4等）与前列腺癌（PCa）的侵略性和/或去势抵抗性PCa（CRPC）的发展有关，但这种现象背后的分子原因是否可能与尚未探讨负责剪接过程的细胞机械失调[剪接体成分（SCs）和剪接因子（SFs）]。方法在两个队列的PCa样本中测量了43个关键SC和SF的表达水平：1）临床定位的福尔马林固定石蜡包埋的PCa样本（n = 84），以及2）攻击性强的新鲜获得的PCa-样本（n = 42）。发现与非肿瘤相邻区域相比，在PCa中发现了剪接机器多个组件（即7个SC / 19个SF）表达的严重失调。值得注意的是，SNRNP200，SRSF3和SRRM1（mRNA和/或蛋白质）的过表达与相关的临床（例如格里森评分，T阶段，转移，生化复发等）和分子（例如AR-v7表达）的攻击性参数有关在PCa样本中。在正常前列腺细胞（PNT2）和PCa细胞（LNCaP / 22Rv1 / PC-3 /中，进行功能（细胞增殖/迁移）和机制[基因表达（qPCR）和蛋白质水平（western-blot）]检测DU145细胞系）响应SNRNP200，SRSF3和/或SRRM1沉默（使用特定的siRNA）显示，通过调节关键致癌SVs表达水平（例如AR-v7 / PKM2 / XBP1s）和致癌信号通路的改变，PCa细胞的增殖/迁移速率总体下降。 （例如p-AKT / p-JNK）。解释这些结果表明，剪接体在PCa中发生了巨大变化，其中SNRNP200，SRSF3和SRRM1可以代表PCa和CRPC的有吸引力的新颖诊断/预后和治疗靶标。