Activated yes-associated protein accelerates cell cycle, inhibits apoptosis, and delays senescence in human periodontal ligament stem cells

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).

Int J Med Sci 2018, Vol 15 Ivyspring International Publisher 1241 International Journal of Medical Sciences 2018; 15(11): 1241-1250 doi: 10.7150/ijms.25115 Research Paper Activated Yes-Associated Protein Accelerates Cell Cycle, Inhibits Apoptosis, and Delays Senescence in Human Periodontal Ligament Stem Cells Linglu Jia1,3*, Weiting Gu2*, Yunpeng Zhang1,3, Baoqi Jiang 1,3, Xu Qiao4, Yong Wen1,3 * School of Stomatology, Shandong University, Jinan, China Department of Obstetrics and Gynecology, Qilu hospital of Shandong University, Jinan, China Shandong provincial key laboratory of oral tissue regeneration, Jinan, China School of Control Science and Engineering, Shandong University, Jinan, China co-first authors: These two authors contributed equally to this work and should be considered as co-first authors  Corresponding authors: Yong Wen (wenyong@sdu.edu.cn),No 44-1, Wenhua Xi Road, Jinan, Shandong, 250012 P.R China and Xu Qiao (qiaoxu@sdu.edu.cn), Jingshi Road 17923, Jinan Shandong, 250012 P.R China © Ivyspring International Publisher This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY-NC) license (https://creativecommons.org/licenses/by-nc/4.0/) See http://ivyspring.com/terms for full terms and conditions Received: 2018.01.23; Accepted: 2018.06.28; Published: 2018.07.30 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