Gao Ruru, Luo Qiong, Li Yang, Song Liming, Cai Junnan Stephen, Xiong Ying, Yan Fei, Liu Jianhua
Department of Medical Ultrasound, The Second Affiliated Hospital, School of Medicine, South China University of Technology, Guangzhou 510180, China.
Department of Gastrointestinal Surgery, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University), Shenzhen 518020, China.
Pharmaceutics. 2022 Jun 30;14(7):1382. doi: 10.3390/pharmaceutics14071382.
The epithelial-mesenchymal transition (EMT), a process in which epithelial cells undergo a series of biochemical changes to acquire a mesenchymal phenotype, has been linked to tumor metastasis. Here, we present a novel strategy for knocking out the EMT-related Cdh2 gene, which encodes N-cadherin through CRISPR/Cas9-mediated gene editing by an ultrasound combined with biosynthetic nanobubbles (Gas Vesicles, GVs). Polyethyleneimine were employed as a gene delivery vector to deliver sgRNA into 4T1 cells that stably express the Cas9 protein, resulting in the stable Cdh2 gene- knockout cell lines. The Western blotting assay confirmed the absence of an N-cadherin protein in these Cdh2 gene-knockout 4T1 cell lines. Significantly reduced tumor cell migration was observed in the Cdh2 gene-knockout 4T1 cells in comparison with the wild-type cells. Our study demonstrated that an ultrasound combined with GVs could effectively mediate CRISPR/Cas9 gene editing of a Cdh2 gene to inhibit tumor invasion and metastasis.
上皮-间质转化（EMT）是上皮细胞经历一系列生化变化以获得间质表型的过程，与肿瘤转移有关。在此，我们提出了一种通过超声联合生物合成纳米气泡（气体囊泡，GVs）进行CRISPR/Cas9介导的基因编辑来敲除与EMT相关的Cdh2基因（该基因编码N-钙黏蛋白）的新策略。使用聚乙烯亚胺作为基因递送载体，将sgRNA递送至稳定表达Cas9蛋白的4T1细胞中，从而获得稳定的Cdh2基因敲除细胞系。蛋白质免疫印迹分析证实这些Cdh2基因敲除的4T1细胞系中不存在N-钙黏蛋白。与野生型细胞相比，在Cdh2基因敲除的4T1细胞中观察到肿瘤细胞迁移显著减少。我们的研究表明，超声联合GVs可有效介导Cdh2基因的CRISPR/Cas9基因编辑，以抑制肿瘤侵袭和转移。
