Gogoi Radhika P, Galoforo Sandra, Fox Alexandra, Morris Colton, Ramos Harry, Gogoi Vir K, Chehade Hussein, Adzibolosu Nicholas K, Shi Chenjun, Zhang Jitao, Tedja Roslyn, Morris Robert, Alvero Ayesha B, Mor Gil
Karmanos Cancer Institute, Wayne State University, 4100 John R St, Detroit, MI 48202, USA.
C.S. Mott Center for Human Growth and Development, Department of Obstetrics and Gynecology, Wayne State University, Detroit, MI 48202, USA.
Cancers (Basel). 2023 Oct 2;15(19):4834. doi: 10.3390/cancers15194834.
Epithelial-mesenchymal transition (EMT) is a biological process where epithelial cells lose their adhesive properties and gain invasive, metastatic, and mesenchymal properties. Maintaining the balance between the epithelial and mesenchymal stage is essential for tissue homeostasis. Many of the genes promoting mesenchymal transformation have been identified; however, our understanding of the genes responsible for maintaining the epithelial phenotype is limited. Our objective was to identify the genes responsible for maintaining the epithelial phenotype and inhibiting EMT.
RNA seq was performed using an vitro model of EMT. CTGF expression was determined via qPCR and Western blot analysis. The knockout of CTGF was completed using the CTGF sgRNA CRISPR/CAS9. The tumorigenic potential was determined using NCG mice.
The knockout of CTGF in epithelial ovarian cancer cells leads to the acquisition of functional characteristics associated with the mesenchymal phenotype such as anoikis resistance, cytoskeleton remodeling, increased cell stiffness, and the acquisition of invasion and tumorigenic capacity.
We identified CTGF is an important regulator of the epithelial phenotype, and its loss is associated with the early cellular modifications required for EMT. We describe a novel role for CTGF, regulating cytoskeleton and the extracellular matrix interactions necessary for the conservation of epithelial structure and function. These findings provide a new window into understanding the early stages of mesenchymal transformation.
上皮-间质转化（EMT）是一个生物学过程，在此过程中上皮细胞失去其黏附特性，获得侵袭性、转移性和间质特性。维持上皮和间质阶段之间的平衡对于组织内稳态至关重要。许多促进间质转化的基因已被确定；然而，我们对负责维持上皮表型的基因的了解有限。我们的目标是确定负责维持上皮表型并抑制EMT的基因。
使用EMT的体外模型进行RNA测序。通过qPCR和蛋白质免疫印迹分析确定结缔组织生长因子（CTGF）的表达。使用CTGF sgRNA CRISPR/CAS9完成CTGF的敲除。使用NCG小鼠确定致瘤潜力。
上皮性卵巢癌细胞中CTGF的敲除导致获得与间质表型相关的功能特征，如失巢凋亡抗性、细胞骨架重塑、细胞硬度增加以及侵袭和致瘤能力的获得。
我们确定CTGF是上皮表型的重要调节因子，其缺失与EMT所需的早期细胞修饰有关。我们描述了CTGF的一个新作用，即调节维持上皮结构和功能所需的细胞骨架和细胞外基质相互作用。这些发现为理解间质转化的早期阶段提供了一个新窗口。
