Splice-Variant Knock-Out of TGF$\beta$ Receptors Perturbates the Proteome of Ovarian Carcinoma Cells.


Liora Jacobs Catane, Ofra Moshel, Yoav Smith, Ben Davidson, and Reuven Reich. 2021. “Splice-Variant Knock-Out of TGF$\beta$ Receptors Perturbates the Proteome of Ovarian Carcinoma Cells.” International journal of molecular sciences, 22, 23.


The aim of this study was to analyze the biological role of different transforming growth factor-$\beta$ (TGF$\beta$) receptor splice variants in ovarian carcinoma (OC). Specific receptor variant knockouts (KO) were prepared using the CRISPR/Cas9 genome editing system in two OC cell lines, T$\beta$RI variant 1 (T$\beta$RIv1) KO in ES-2 cells and T$\beta$RII variant 1 (T$\beta$RIIv1) KO in OVCAR-8 cells. Control and KO cells were compared by proteomic analysis, functional tests, analysis of epithelial-mesenchymal transition (EMT) drivers, and Western blot of signaling proteins. Proteomic analysis revealed significant changes in protein pathways in the KO cells. T$\beta$RIv1 KO resulted in a significant reduction in both cellular motility and invasion, while T$\beta$RIIv1 KO significantly reduced cellular motility and increased Reactive Oxygen Species (ROS) production. Both receptor variant KOs reduced MET protein levels. Of the EMT drivers, a significant decrease in TWIST protein expression, and increase in SNAIL protein and MALAT1 mRNA levels were observed in the T$\beta$RIIv1 KO compared to control. A significant decrease in JNK1 and JNK2 activation was found in the T$\beta$RIv1 KO compared to control cells. These findings provide new insight regarding the biological role of the TGF$\beta$ receptor variants in the biology and potentially the progression of OC.