International Journal of Medical Sciences

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19 October 2017

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Int J Med Sci 2017; 14(12):1231-1240. doi:10.7150/ijms.20504

Research Paper

Knockdown of Yes-Associated Protein Induces the Apoptosis While Inhibits the Proliferation of Human Periodontal Ligament Stem Cells through Crosstalk between Erk and Bcl-2 Signaling Pathways

Yong Wen1, 2, Yawen Ji1, 2, Yunpeng Zhang1, 2, Baoqi Jiang1, 2, Cuizhu Tang1, 2, Qi Wang1, 2, Xiyan Chen1, 2, Linglu Jia1, 2, Weiting Gu3✉, Xin Xu1, 2✉

1. School of Stomatology, Shandong University, Jinan, China;
2. Shandong provincial key laboratory of oral tissue regeneration , Jinan, China;
3. Qilu hospital of Shandong University, Jinan, China.

Abstract

Objective: The purpose of this study was to provide an insight into the biological effects of knockdown Yes-associated protein (YAP) on the proliferation and apoptosis of human periodontal ligament stem cells (h-PDLSCs). Methods: Immunofluorescence and Western blot were used to evaluate Hippo-YAP signaling expression level. Enhanced green fluorescence protein lentiviral vector was constructed to down-regulate YAP in h-PDLSCs. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot were used to detect the interfering efficiency of YAP expression. The proliferation activity was detected by EdU staining. Analysis of apoptosis in h-PDLSCs was done through Annexin V-APC staining, while cell cycle analysis was detected by flow cytometry. Cellular senescence was analyzed by β-galactosidase activity detection. The expression of elements in signaling pathways related with proliferation and apoptosis was detected by Western blot. Results: YAP was located in nucleus and cytoplasm. After the lentivirus transfection, the expression of YAP mRNA and protein was significantly reduced (P<0.001). When YAP was knocked down, the proliferation activity of h-PDLSCs was inhibited; the early & late apoptosis rates increased; the proportion of cells in G1 phases increased (P<0.05), while that in G2 and S phase decreased (P<0.05); cellular senescence was accelerated (P<0.01); ERK and its target proteins P-P90RSK and P-MEK were reduced while Bcl-2 family members increased. Conclusion: Knockdown of YAP inhibits the proliferation activity and induces apoptosis of h-PDLSCs with the involvement of Hippo pathway and has a crosstalk between Erk and Bcl-2 signaling pathways.

Keywords: Yes-associated protein (YAP), human periodontal ligament stem cells (h-PDLSCs), proliferation, apoptosis.

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How to cite this article:
Wen Y, Ji Y, Zhang Y, Jiang B, Tang C, Wang Q, Chen X, Jia L, Gu W, Xu X. Knockdown of Yes-Associated Protein Induces the Apoptosis While Inhibits the Proliferation of Human Periodontal Ligament Stem Cells through Crosstalk between Erk and Bcl-2 Signaling Pathways. Int J Med Sci 2017; 14(12):1231-1240. doi:10.7150/ijms.20504. Available from http://www.medsci.org/v14p1231.htm