Roles of Yes-associated protein in epidermal stem cell differentiation after ionizing radiation
10.3867/j.issn.1000-3002.2024.07.004
- VernacularTitle:Yes相关蛋白在电离辐射后表皮干细胞分化中的作用
- Author:
Libin WANG
1
,
2
;
Junfei CHEN
;
Fang YUAN
;
Jingze WANG
;
Lu LIU
;
Xuewu WANG
;
Zengqiang YUAN
Author Information
1. 南华大学衡阳医学院,军事医学研究院研究生协作培养基地,湖南 衡阳 421001
2. 军事医学研究院,北京 100850
- Keywords:
Yes-associated protein;
ionizing radiation;
epidermal stem cell;
cell differentiation
- From:
Chinese Journal of Pharmacology and Toxicology
2024;38(7):511-516
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To explore the role of Yes-associated protein(YAP)in epidermal stem cell(EPSC)differentiation after ionizing radiation(IR).METHODS ① A punch was used to induce skin injuries on the back of mice.The IR group received localized irradiation with 60 Co γ-rays,while the normal control group did not.Samples were collected at 0,1,3,6,9,and 12 d for RNA and protein extraction.Western blotting was used to detect changes in YAP protein expressions during wound healing.Real-time quantitative polymerase chain reaction(RT-qPCR)was performed to assess the mRNA levels of Yap and its downstream target genes,connective tissue growth factor(Ctgf),and cysteine-rich protein 61(Cyr61).② EPSCs were exposed to 60 Co γ at a dose of 4 or 8 Gy,while the control group was not irradiated.Cells were collected to detect the levels of YAP protein via Western blotting.Cells were collected at 4,12,24,and 36 h post-IR to assess the levels of YAP mRNA by RT-qPCR.③ Short hairpin RNA(shRNA)was used to establish stable YAP knockdown cell lines,and the knockdown efficiency of sh YAP was verified by Western blotting.RT-qPCR was then performed to detect the impact of YAP knockdown on mRNA levels of K1 and K10 after IR.RESULTS① Compared with the control group,the YAP protein level in the IR group during wound healing was significantly reduced(P<0.05,P<0.01),so were the mRNA levels of Yap and its downstream target genes Ctgf and Cyr61(P<0.05,P<0.01).② Compared to the cell control group,the mRNA and protein levels of YAP in the IR group cells were significantly reduced(P<0.01).③ In the sh YAP cells,the YAP protein level was significantly reduced(P<0.01).Furthermore,the mRNA levels of K1 and K10 were significantly decreased after IR in sh YAP cells(P<0.01).CONCLUSION YAP can regulate EPSC differentiation in wound healing after IR.
