PMID- 31065276
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20220408
IS  - 1687-966X (Print)
IS  - 1687-9678 (Electronic)
VI  - 2019
DP  - 2019
TI  - Effect of YAP on an Immortalized Periodontal Ligament Stem Cell Line.
PG  - 6804036
LID - 10.1155/2019/6804036 [doi]
LID - 6804036
AB  - OBJECTIVE: To establish an immortalized human periodontal ligament stem cell line 
      (hPDLSC) and investigate whether and how YAP mediates the establishment of the 
      stem cell line. METHODS: Primary hPDLSCs were cultured and transfected with 
      lentivirus containing the telomerase reverse transcriptase (TERT) gene. The 
      expression of TERT was detected via the polymerase chain reaction (PCR) and 
      real-time quantitative PCR (RT-PCR). Flow cytometry was employed to detect 
      surface markers of hPDLSCs and TERT-hPDLSCs. The cell counting kit-8 (CCK-8) and 
      5-ethynyl-2'-deoxyuridine (EdU) methods were used to examine the proliferation 
      ability of the cells. Flow cytometry and TUNEL staining were employed to examine 
      the cell apoptosis rate. The beta-galactosidase staining assay was used to assess 
      the rate of cell senescence. The osteogenic differentiation ability of the cells 
      was detected via alkaline phosphatase (ALP) staining and Alizarin red staining 
      assays. BALB/c mice were employed to determine the tumorigenicity of 
      TERT-hPDLSCs. The expression levels of YAP and other proteins in the Hippo 
      signaling pathway were detected by Western blotting. Verteporfin was used to 
      inhibit the binding of YAP to the downstream target gene TEAD. RESULTS: 
      TERT-hPDLSCs showed stable high expression of TERT, even at the thirtieth passage 
      after transfection with lentivirus containing the TERT gene. Compared with 
      primary hPDLSCs, TERT-hPDLSCs exhibited a stronger proliferation ability and 
      lower cell apoptosis and senescence rates while maintaining the same osteogenetic 
      differentiation ability as primary hPDLSCs. The transfection of hPDLSCs with 
      lentivirus containing the TERT gene did not lead to tumorigenesis in nude mice. 
      The Hippo signaling pathway was inactivated in TERT-hPDLSCs compared to hPDLSCs. 
      When treated with verteporfin, the proliferation of TERT-hPDLSCs decreased, while 
      the apoptosis and senescence rates of these cells increased. However, 
      TERT-hPDLSCs still showed a stronger proliferation ability and lower cell 
      apoptosis and senescence rates than hPDLSCs treated with verteporfin at the same 
      concentration. CONCLUSIONS: Overexpression of TERT in hPDLSCs resulted in the 
      successful establishment of an immortalized periodontal ligament stem cell line. 
      TERT may regulate the biological characteristics of hPDLSCs through the Hippo/YAP 
      signaling pathway. hPDLSCs could be a feasible resource for stem cell research 
      and a promising resource for stem cell therapy.
FAU - Chen, Xiyan
AU  - Chen X
AD  - Shandong Provincial Key Laboratory of Oral Tissue Regeneration, School of 
      Stomatology, Shandong University, Jinan, Shandong Province, China.
AD  - School of Stomatology, Shandong University, Jinan, Shandong Province, China.
FAU - Wang, Qi
AU  - Wang Q
AD  - Shandong Provincial Key Laboratory of Oral Tissue Regeneration, School of 
      Stomatology, Shandong University, Jinan, Shandong Province, China.
AD  - School of Stomatology, Shandong University, Jinan, Shandong Province, China.
AD  - Ningbo Stomatology Hospital, Zhejiang, China.
FAU - Gu, Ke
AU  - Gu K
AD  - Shandong Provincial Key Laboratory of Oral Tissue Regeneration, School of 
      Stomatology, Shandong University, Jinan, Shandong Province, China.
AD  - School of Stomatology, Shandong University, Jinan, Shandong Province, China.
FAU - Li, Aonan
AU  - Li A
AD  - Shandong Provincial Key Laboratory of Oral Tissue Regeneration, School of 
      Stomatology, Shandong University, Jinan, Shandong Province, China.
AD  - School of Stomatology, Shandong University, Jinan, Shandong Province, China.
FAU - Fu, Xucheng
AU  - Fu X
AD  - Shandong Provincial Key Laboratory of Oral Tissue Regeneration, School of 
      Stomatology, Shandong University, Jinan, Shandong Province, China.
AD  - School of Stomatology, Shandong University, Jinan, Shandong Province, China.
FAU - Wang, Ying
AU  - Wang Y
AD  - Shandong Provincial Key Laboratory of Oral Tissue Regeneration, School of 
      Stomatology, Shandong University, Jinan, Shandong Province, China.
AD  - School of Stomatology, Shandong University, Jinan, Shandong Province, China.
FAU - Gu, Weiting
AU  - Gu W
AD  - Qilu Hospital of Shandong University, Jinan, Shandong Province, China.
FAU - Wen, Yong
AU  - Wen Y
AUID- ORCID: 0000-0002-3446-0959
AD  - Shandong Provincial Key Laboratory of Oral Tissue Regeneration, School of 
      Stomatology, Shandong University, Jinan, Shandong Province, China.
AD  - School of Stomatology, Shandong University, Jinan, Shandong Province, China.
LA  - eng
PT  - Journal Article
DEP - 20190401
PL  - United States
TA  - Stem Cells Int
JT  - Stem cells international
JID - 101535822
PMC - PMC6466850
EDAT- 2019/05/09 06:00
MHDA- 2019/05/09 06:01
PMCR- 2019/04/01
CRDT- 2019/05/09 06:00
PHST- 2018/04/23 00:00 [received]
PHST- 2018/11/08 00:00 [revised]
PHST- 2018/12/16 00:00 [accepted]
PHST- 2019/05/09 06:00 [entrez]
PHST- 2019/05/09 06:00 [pubmed]
PHST- 2019/05/09 06:01 [medline]
PHST- 2019/04/01 00:00 [pmc-release]
AID - 10.1155/2019/6804036 [doi]
PST - epublish
SO  - Stem Cells Int. 2019 Apr 1;2019:6804036. doi: 10.1155/2019/6804036. eCollection 
      2019.