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    Modulation of YAP1 expression and other related signaling pathways in HepG2 cells by extracellular matrix, contact inhibition and cell density
    (American Association for the Study of Liver Diseases, 2022) Aktas, Ranan; Akbulut, Zeynep; Maraş, Hatice; Çakıl Dönmez, Yaprak; Kayalı, Damla; Karagoz Koroglu, Ayca; Sitar, Mustafa Erinc; Kısakol, Batuhan; Baysan, Mehmet
    Background: Hippo-YAP pathway is a key modulator in liver development, regeneration and the metabolism . Cell density, cell contact, the actin cytoskeleton, and extracellular matrix (ECM) composition have been shown to be involved in regulation of this pathway . The intervention of this pathway in hepatocellular carcinoma remains a central focus . The study aimed to examine how YAP1 integrate mechanical cues with the response to signal transduction pathways and to multiple aspects of cell behavior, proliferation, protein/lipid secretion, apoptosis/necrosis, adhesion, and stemness . Methods: HepG2 cells were cultured on uncoated (1), or Type I collagen (2), standard matrigel (3) or high concentration-matrigel coated surfaces (4) for 6 weeks . The cells were examined at 60% or 100%confluency, and at the end of each week by qRT-PCR, confocal microscopy, MTT, ELISA, and flow cytometry. Expression of 84 genes including YAP1, cell adhesion molecules (Inhibin, Cadherin, Na-K ATPase), apoptotic/necrotic changes, viability, protein/ lipid secretion, and existence of 7 different cancer stem cells were compared with Spearmen correlation test . Results: YAP1 gene expression changed significantly in Group1. Contact inhibition with 100% confluency caused deacrease at YAP1 in all groups . It was always low in group4 when compared with the others . Strong correlation between the YAP1, Actin-beta, and most of the genes in TGFB, AKT-P13 and RAS/RAF/MAPK pathways were clear (Graph I) . Positive correlation between cholesterol /LDL/HDL secretion; negative correlation between protein secretion, necrosis and CD133 (+) cells were notable . Conclusion: Herein, we described for the first time how liver cancer cells translate complex and dynamic changes at the external cues into signaling conduits that impact on YAP and other related gene expressions as well as on metabolic and morphological features of the cells . The study demonstrated the striking changes through the receptor of integrin alpha2beta1 that is stimulated by type I collagen . Changes in group 3 and 4 suggest that growth factors in stroma is another important factor at the response related to YAP1 expression. To our knowledge, this is the first study demonstrates Actin and YAP1 interaction at the gene expression level .