Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Pathology, Peking University Cancer Hospital & Institute, Beijing, China.
Department of Biochemistry and Biophysics, Beijing Key Laboratory of Protein Posttranslational Modifications and Cell Function, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, China.
Cell Death Dis. 2023 Jul 14;14(7):431. doi: 10.1038/s41419-023-05951-5.
Pancreatic cancer is a leading cause of cancer death due to its early metastasis and limited response to the current therapies. Metastasis is a complicated multistep process, which is determined by complex genetic alterations. Despite the identification of many metastasis-related genes, distinguishing the drivers from numerous passengers and establishing the causality in cancer pathophysiology remains challenging. Here, we established a high-throughput and piggyBac transposon-based genetic screening platform, which enables either reduced or increased expression of chromosomal genes near the incorporation site of the gene search vector cassette that contains a doxycycline-regulated promoter. Using this strategy, we identified YWHAZ as a key regulator of pancreatic cancer metastasis. We demonstrated that functional activation of Ywhaz by the gene search vector led to enhanced metastatic capability in mouse pancreatic cancer cells. The metastasis-promoting role of YWHAZ was further validated in human pancreatic cancer cells. Overexpression of YWHAZ resulted in more aggressive metastatic phenotypes in vitro and a shorter survival rate in vivo by modulating epithelial-to-mesenchymal transition. Hence, our study established a high-throughput screening method to investigate the functional relevance of novel genes and validated YWHAZ as a key regulator of pancreatic cancer metastasis.
胰腺癌是癌症死亡的主要原因，因为它早期转移且对当前疗法的反应有限。转移是一个复杂的多步骤过程，由复杂的遗传改变决定。尽管已经鉴定出许多与转移相关的基因，但区分驱动基因和众多乘客，并在癌症病理生理学中建立因果关系仍然具有挑战性。在这里，我们建立了一种高通量的基于 piggyBac 转座子的遗传筛选平台，该平台能够降低或增加基因搜索载体盒整合部位附近的染色体基因的表达，该载体盒包含一个由强力霉素调控的启动子。利用该策略，我们鉴定出 YWHAZ 是胰腺癌转移的关键调节因子。我们证明，基因搜索载体的功能性激活导致小鼠胰腺癌细胞转移能力增强。YWHAZ 的转移促进作用在人胰腺癌细胞中得到了进一步验证。YWHAZ 的过表达通过调节上皮-间充质转化，导致体外更具侵袭性的转移表型和体内存活率降低。因此，我们的研究建立了一种高通量筛选方法来研究新基因的功能相关性，并验证了 YWHAZ 是胰腺癌转移的关键调节因子。
