Activated Yes-Associated Protein Accelerates Cell Cycle, Inhibits Apoptosis, and Delays Senescence in Human Periodontal Ligament Stem Cells

Abstract

Objectives: To provide insight into the biological effects of activated Yes-associated protein (YAP) on the proliferation, apoptosis, and senescence of human periodontal ligament stem cells (h-PDLSCs).Methods: h-PDLSCs were isolated by the limiting dilution method, and their surface markers were quantified by flow cytometry. Enhanced green fluorescence protein (EGFP)-labeled lentiviral vector was used to activate YAP in h-PDLSCs, then qRT-PCR and Western blotting were used to evaluate the expression level of YAP. Immunofluorescence was used to detect the location of YAP in h-PDLSCs. The proliferation activity was detected by cell counting kit-8 (CCK-8) and 5-ethynyl-2'-deoxyuridine (EdU), and the cell cycle was determined by flow cytometry. Apoptosis was analyzed by Annexin V-APC staining. Cell senescence was detected by β-galactosidase staining. Proteins in ERK, Bcl-2, and p53 signaling pathways were detected by Western blotting.Results: h-PDLSCs were isolated successfully and were positive for human mesenchymal stem cell surface markers. After YAP was activated by lentiviral vector, the mRNA and protein of YAP were highly expressed, and more YAP translocated into the nucleus. When YAP was overexpressed in h-PDLSCs, proliferation activity was improved; early and late apoptosis rates decreased (P<0.05); the proportion of cells in G2/M phases increased (P<0.05), while that in G0/G1 phase decreased (P<0.05); cellular senescence was delayed (P<0.01); the expression of P-MEK, P-ERK, P-P90RSK and P-Msk increased, while the expression of Bcl-2 family members (Bak, Bid and Bik) decreased.Conclusions: Activated YAP promotes proliferation, inhibits apoptosis, and delays senescence of h-PDLSCs. The Hippo-YAP signaling pathway can influence ERK and Bcl-2 signaling pathways.

Revisiting the Hayflick Limit: Insights from an Integrated Analysis of Changing Transcripts, Proteins, Metabolites and Chromatin

“…Following increases in DNA accessiblity, expression of newly opened TEAD1/YAP1/SMAD target genes cement FMT transition by promoting fibrosis and a myofibroblast-like, ECM-secreting state. This model is supported by the synergy between TEAD1 motifs and NAD domains we report and reconciles conflicting reports that YAP1/TEAD1 inhibition can both prevent and promote senescence [125, 84, 126, 127]. Basically, the functional consequences of YAP1/TEAD1 inhibition will depend on the epigenetic organization of the cells used.…”

Geranylgeraniol prevents zoledronic acid-mediated reduction of viable mesenchymal stem cells via induction of Rho-dependent YAP activation

“…The present mechanism of action study provided the first evidence supporting that GGOH-prevented MSC death from ZA cytotoxicity was at least partly mediated through Rho-dependent activation of YAP. Rho subfamily proteins and YAP have been shown to play important roles in viability, proliferation and stemness of MSCs [ 20 , 21 , 23 , 24 ].…”

“…The present results demonstrated that GGOH-mediated YAP activation reversed ZA-induced cell cycle arrest by stimulating cell cycle progression and promoting more cells to enter S phase and G2/M phase, as previously reported for the role of YAP in the induction of cell cycle progression in chondrocytes [ 53 ]. Moreover, YAP has been shown to stimulate proliferation and inhibit apoptosis of stem cells [ 21 , 22 ]. However, it is also possible that in this context, other downstream effectors of Rho can also mediate cell division and proliferation of MSCs.…”

“…YAP activity is critical for whole organ growth, for the expansion of progenitor cells during tissue regeneration, and for cell proliferation [ 20 ]. It has been shown that YAP promotes proliferation, impedes apoptosis and delays the senescence of stem cells derived from human periodontal ligament cells [ 21 ]. A previous study further suggested a critical role of YAP in maintaining stem cell pluripotency [ 22 ].…”

Key Markers and Epigenetic Modifications of Dental-Derived Mesenchymal Stromal Cells

“…As a Hippo signaling transcriptional coactivator, YAP plays pivotal roles in MSCs fate and organ size control [134]. Jia et al discovered that activated YAP promotes proliferation, accelerates the cell cycle, inhibits apoptosis, and delays senescence in human PDLSCs [135]. Knockdown of YAP inhibits the proliferation activity and induces apoptosis of human PDLSCs with the involvement of the Hippo pathway and shows crosstalk with the Erk and Bcl-2 signaling pathways 7 Stem Cells International [136].…”