Collak Filiz Kisaayak, Demir Ummuhan, Sagir Fatma
Faculty of Engineering and Natural Sciences, Molecular Biology and Genetics Department, Istanbul Medeniyet University, 34700, Uskudar, Istanbul, Turkey.
Pathol Oncol Res. 2020 Apr;26(2):867-876. doi: 10.1007/s12253-019-00634-z. Epub 2019 Mar 11.
The Yes Associated Protein 1 (YAP1) is a transcriptional cofactor negatively regulated by Hippo Pathway. The dysregulation of the pathway has been shown to have a role in tumorigenesis and metastasis in several cancers including prostate cancer (PCa). In this study, YAP1 expression was upregulated in the whole cell lysates and cytoplasmic/nuclear extracts of AR negative (PC3) compared to AR positive (LNCaP) prostate cancer cell lines and primary prostate epithelial cells (PrePEC). pYAP1 expression elevated in LNCaP compared to PC3 and PrePEC in whole cell lysates and cytoplasmic extracts. The treatment of LNCaP and PC3 with YAP1-targeting siRNA oligonucleotides (YAP1 siRNA) significantly reduced their proliferation in vitro. Furthermore, treatment with YAP1 siRNA diminished the clonogenicity, anchorage-independent growth on soft agar, migration and invasion of PC3 cells. Co-IP/WB experiments revealed that YAP1 and AR formed a complex and ChIP/PCR results confirmed that YAP1 was bound to androgen response elements (ARE) core region of the prostate specific antigen (PSA) promoter. The loss of function experiments in LNCaP and PC3 revealed that YAP1 regulates proliferation, colony formation as well as anchorage-independent growth and potentially plays an important role in migration and invasion. Finally, analysis of publicly available data sets indicated that LNCaP had no YAP1 copy number alteration whereas PC3 had gain of YAP1 which was also reflected as an increase in the mRNA level. Moreover, YAP1 copy number gain and elevated YAP1 mRNA expression were detected in clinical samples analyzed in publicly available data sets. Taken together, these results suggested that YAP1 has a role in PCa tumorigenesis.
Yes相关蛋白1（YAP1）是一种转录辅因子，受Hippo通路负调控。该通路的失调已被证明在包括前列腺癌（PCa）在内的几种癌症的肿瘤发生和转移中起作用。在本研究中，与雄激素受体（AR）阳性（LNCaP）前列腺癌细胞系和原代前列腺上皮细胞（PrePEC）相比，AR阴性（PC3）的全细胞裂解物以及细胞质/细胞核提取物中YAP1表达上调。在全细胞裂解物和细胞质提取物中，与PC3和PrePEC相比，LNCaP中的pYAP1表达升高。用靶向YAP1的小干扰RNA寡核苷酸（YAP1 siRNA）处理LNCaP和PC3可显著降低其体外增殖。此外，用YAP1 siRNA处理可降低PC3细胞的克隆形成能力、在软琼脂上的非锚定依赖性生长、迁移和侵袭能力。免疫共沉淀/蛋白质印迹（Co-IP/WB）实验表明YAP1和AR形成复合物，染色质免疫沉淀/聚合酶链反应（ChIP/PCR）结果证实YAP1与前列腺特异性抗原（PSA）启动子的雄激素反应元件（ARE）核心区域结合。在LNCaP和PC3中的功能缺失实验表明，YAP1调节增殖、集落形成以及非锚定依赖性生长，并可能在迁移和侵袭中起重要作用。最后，对公开可用数据集的分析表明，LNCaP没有YAP1拷贝数改变，而PC3有YAP1拷贝数增加，这也反映为mRNA水平的升高。此外，在公开可用数据集中分析的临床样本中检测到YAP1拷贝数增加和YAP1 mRNA表达升高。综上所述，这些结果表明YAP1在PCa肿瘤发生中起作用。
