The study of molecular mechanism of regulation of IL-10 on proliferation and differentiation of HaCaT cells
10.19405/j.cnki.issn1000-1492.2023.06.002
- Author:
Xueli Yin
1
;
Bo Jia
2
;
Li Liu
3
;
Mingcong Li
4
,
5
;
Jun Zhang
6
;
Zhen Yang
6
;
Hongmei Bai
6
;
Weikang Hu
6
;
Sumei Zhang
6
;
Shengquan Zhang
6
Author Information
1. Functional Experiment Center，Anhui Medical University，Hefei 230032
2. Xiangyang Institute for Food and Drug Control，Xiangyang 441000
3. Center for Scientific Research，Anhui Medical University，Hefei 230032
4. Dept of Pathology，The Second People'
5. s Hospital of Hefei，Hefei 230011
6. Dept of Biochemistry and Molecular Biology， Anhui Medical University，Hefei 230032
- Publication Type:Journal Article
- Keywords:
HaCaT cells;
cell proliferation;
IL-10;
differentiation markers;
signaling pathways
- From:
Acta Universitatis Medicinalis Anhui
2023;58(6):890-895
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of interleukin ( IL) -10 on the proliferation of HaCaT cells and CaCl2 induced expression of differentiation markers and its possible molecular mechanisms．
Methods:HaCaT cells were treated with various concentrations of IL-10 (0，3，10，30 ng / ml) for different time (0，24，48，72 h) ，cell proliferation was measured using MTS，and cell cycle was determined by flow cytometry．HaCaT cells were pretreated with IL-10 (final concentration 10 ng / ml) for 1 h，then incubated with or without CaCl2 (final concentration 1. 2 mmol / L) for 24，48，72 h ，Western blot was performed to detect the effect of IL-10 on the expression of HaCaT keratinocyte differentiation markers．After pretreatment of HaCaT cells with PD98059，an inhibitor of mitogen-activated kinase-ERK1 /2，and LY294002，an inhibitor of phosphatidylinositol kinase-serine / threonine kinase (PI3K-AKT) ，the total RNA and proteins were extracted separately，real time quantitative polymerase chain reaction (RT-qPCR) and Western blot were used to examine the influence of IL-10 on the expression of differentiation markers (Keratin1，Keratin5，Involucrin) .
Results :MTS results revealed that IL-10 (30 ng / ml and lower doses) did not alter the proliferation of HaCaT cells in 72 h．Flow cytometry analysis demonstrated that IL-10 had no significant influence on cell cycle progression．The results of Western blot showed that IL-10 upregulated the expression of differentiation markers Involucrin，while there was no significant effect on Keratin1 and Keratin5 ．Mechanism research analysis demonstrated that IL-10 could activate ERK1 /2 and AKT ，increase their phosphorylation levels ; RT-qPCR and Western blot results showed that PD98059 and LY294002 partially blocked IL-10 induced Involucrin expression．
Conclusion:At a particular concentration range，IL-10 has little effect on HaCaT proliferation ，but it partially upregulates the expression of differentiation marker Involucrin via the MAPKs-ERK1 /2 and PI3K-AKT pathways.
- Full text:2024081616371240229IL-10调控HaCaT细胞增殖及分化的分子机制_尹雪莉.pdf
