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.
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.
复制标题
Yes 相关蛋白的敲低通过 Erk 和 Bcl-2 信号通路之间的串扰诱导细胞凋亡，同时抑制人牙周膜干细胞的增殖
DOI：
10.7150/ijms.20504
复制
发表时间：
2017
影响因子：
3.6
通讯作者：
Xu X
中科院分区：
文献类型：
--
作者：
Wen Y;Ji Y;Zhang Y;Jiang B;Tang C;Wang Q;Chen X;Jia L;Gu W;Xu X
目的：研究yes相关蛋白（YAP）敲低对人牙周韧带干细胞（h-PDLSCs）增殖和凋亡的生物学影响。方法：采用免疫荧光法和Western blot法检测大鼠Hippo-YAP信号表达水平。构建增强型绿色荧光蛋白慢病毒载体，下调h-PDLSCs中的YAP。采用实时定量逆转录聚合酶链反应（qRT-PCR）和Western blot检测对YAP表达的干扰效率。EdU染色检测细胞增殖活性。Annexin V-APC染色分析h-PDLSCs细胞凋亡，流式细胞术检测细胞周期。采用β-半乳糖苷酶活性检测分析细胞衰老情况。Western blot检测细胞增殖和凋亡相关信号通路中相关元件的表达。结果：YAP位于细胞核和细胞质中。慢病毒转染后，YAP mRNA和蛋白的表达量显著降低（P<0.001）。当YAP被敲除时，h-PDLSCs的增殖活性受到抑制；早期和晚期细胞凋亡率升高；G1期细胞比例增加（P<0.05）， G2、S期细胞比例减少（P<0.05）；细胞衰老加速（P<0.01）；ERK及其靶蛋白P-P90RSK和P-MEK减少，Bcl-2家族成员增加。结论：YAP基因敲低可抑制h-PDLSCs的增殖活性并诱导其凋亡，其参与Hippo通路，且Erk与Bcl-2信号通路之间存在串扰。
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.
登录
查看更多内容
影响因子：
12.4
作者：
Fausti, F.;Di Agostino, S.;Blandino, G.
通讯作者：
Blandino, G.
DOI：
10.1038/nrd4161
发表时间：
2014-01
期刊：
Nature reviews. Drug discovery
影响因子：
--
作者：
通讯作者：
--
影响因子：
64.5
作者：
Alarcón C;Zaromytidou AI;Xi Q;Gao S;Yu J;Fujisawa S;Barlas A;Miller AN;Manova-Todorova K;Macias MJ;Sapkota G;Pan D;Massagué J
通讯作者：
Massagué J
影响因子：
16.6
作者：
Watt, K. I.;Turner, B. J.;Gregorevic, P.
通讯作者：
Gregorevic, P.
影响因子：
16.6
作者：
Lee DH;Park JO;Kim TS;Kim SK;Kim TH;Kim MC;Park GS;Kim JH;Kuninaka S;Olson EN;Saya H;Kim SY;Lee H;Lim DS
通讯作者：
Lim DS