Effects of Notch signaling pathway on proliferation of infantile hemangioma-derived mesenchymal stem cells
10.3760/cma.j.cn114453-20200507-00270
- VernacularTitle:Notch信号通路对婴幼儿血管瘤间充质干细胞增殖的影响
- Author:
Weidong WANG
1
;
Tao HAN
;
Sheng CHEN
;
Yuan WANG
;
Weimin SHEN
Author Information
1. 南京医科大学附属儿童医院烧伤整形科 210008
- Keywords:
Hemangioma;
Cell propliferation;
Mesenchymal stem cells;
Notch
- From:
Chinese Journal of Plastic Surgery
2021;37(5):554-561
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the effects of Notch signaling pathway on proliferation of infantile hemangioma-derived mesenchymal stem cells.Methods:25 specimens from the patients (3 months to 4 years old) were collected at the Department of Burns and Plastic Surgery of the Children’s Hospital of Nanjing Medical University. According to Mulliken classification of IHs, all patients were divided into the proliferating phase group ( n=15), the involuting phase group ( n=10)and 10 normal skin specimens were collected for control. The expressions of Notch components in different phases of hemangioma were assayed by real-time polymerase chain reaction (RT-PCR). Hem-MSCs were isolated from hemangioma in proliferating phase by their select iveadhesion to culture dishes and were indentified by flow cytometry assay.The agonist Jagged1 and inhibitor DAPT of Notch signaling were added into the culture medium, and then the cells were divided into control group, Jagged1 group and DAPT group.After 72 hours, mRNA of Notch1, jagged1 and Hes1 in PCs before and after intervention were detected by RT-PCR. Western blot was performed to detect the protein expression of Notch signaling before and after intervention. Cell proliferation was quantified by cell counting kit-8 (CCK-8) assay. The t test was used to compare the mean of two groups. Statistical significance was set at P<0.05. Results:The expressions of Notch1, Jagged1 and Hes1 in proliferating phase(0.379±0.121, 0.243±0.095, 0.222±0.060) and involuting phase(0.338±0.091, 0.405±0.107, 0.310±0.109) are higher than normal skin(0.111±0.042, 0.065±0.036, 0.074±0.029)( P<0.05). There was no significant difference of Notch1 expression in hemangioma at different stages( t=0.896, P=0.380). The expression of Jagged1 and Hes1 in involuting group is higher than in proliferating phase group( tJagged1=3.895, PJagged1=0.007, tHes1=2.603, PHes1=0.016). The Hem-MSCs were successfully cultured in vitro. These cells expressed typical MSCs markers, CD105, CD90, except for CD34, CD45. Compared with the control group, the expression of Notch1, jagged1 and Hes1gene(1.41±0.37, 2.42±0.39, 1.89±0.35) and protein(0.42±0.25, 1.14±0.32, 0.74±0.35) increased with Jagged1( P<0.05, the expression of Notch1, jagged1 and Hes1 gene(0.58±0.31, 0.56±0.21, 0.43±0.15) and protein(0.19±0.11, 0.62±0.25, 0.12±0.04)decreased by using DAPT( P<0.05). Compared with the control group, Jagged1 can inhibit the cell activityand proliferation of hemangioma-derived mesenchymal stem cells( t=2.447, P=0.034), DAPT can enhance the cell activity and proliferation of hemangioma-derived mesenchymal stem cells( t=3.526, P=0.006). Conclusions:There is an activated Notch signaling pathway in Hem-MSCs. Activation of Notch signaling pathway inhibits proliferation of Hem-MSCs.
