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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):1231-1240IJ

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.

Authors+Show Affiliations

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

Pub Type(s)

Journal Article

Language

eng

PubMed ID

29104479

Citation

Wen, Yong, et al. "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." International Journal of Medical Sciences, vol. 14, no. 12, 2017, pp. 1231-1240.
Wen Y, Ji Y, Zhang Y, et al. 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.
Wen, Y., Ji, Y., Zhang, Y., Jiang, B., Tang, C., Wang, Q., ... Xu, X. (2017). 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. International Journal of Medical Sciences, 14(12), pp. 1231-1240. doi:10.7150/ijms.20504.
Wen Y, et al. 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. PubMed PMID: 29104479.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - 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. AU - Wen,Yong, AU - Ji,Yawen, AU - Zhang,Yunpeng, AU - Jiang,Baoqi, AU - Tang,Cuizhu, AU - Wang,Qi, AU - Chen,Xiyan, AU - Jia,Linglu, AU - Gu,Weiting, AU - Xu,Xin, Y1 - 2017/09/19/ PY - 2017/04/10/received PY - 2017/08/07/accepted PY - 2017/11/7/entrez PY - 2017/11/7/pubmed PY - 2018/6/22/medline KW - Yes-associated protein (YAP) KW - apoptosis. KW - human periodontal ligament stem cells (h-PDLSCs) KW - proliferation SP - 1231 EP - 1240 JF - International journal of medical sciences JO - Int J Med Sci VL - 14 IS - 12 N2 - 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. SN - 1449-1907 UR - https://www.unboundmedicine.com/medline/citation/29104479/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_ L2 - http://www.medsci.org/v14p1231.htm DB - PRIME DP - Unbound Medicine ER -