Establishment of skin photoaging model and preliminary mechanism of senescence in keratinocytes cell line HaCaT

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Establishment of skin photoaging model and preliminary mechanism of senescence in keratinocytes cell line HaCaT

Basic Medicine | 更新时间：2026-01-09

- Establishment of skin photoaging model and preliminary mechanism of senescence in keratinocytes cell line HaCaT
- Journal of Third Military Medical University Vol. 41, Issue 17, Pages: 1649-1655(2019)
- 作者机构：
  
  1.400016 重庆，重庆医科大学:附属第一医院皮肤科
  2.400016 重庆，重庆医科大学:中医药学院
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81573027);the General Program of National Natural Science Foundation of China(81874238);the Natural Science Foundation of Chongqing(CSTC2016jcyjA0430)
- DOI：10.16016/j.1000-5404.201903210
  CLC： R-332;R322.991;R363.122
- Received：25 March 2019，
  
  Revised：2019-05-30，
  
  Published：15 September 2019
- 稿件说明：
移动端阅览
Chuan LIU, Xin HUANG, Ping WANG, et al. Establishment of skin photoaging model and preliminary mechanism of senescence in keratinocytes cell line HaCaT[J]. Journal of Third Military Medical University, 2019, 41(17): 1649-1655.
DOI：

Chuan LIU, Xin HUANG, Ping WANG, et al. Establishment of skin photoaging model and preliminary mechanism of senescence in keratinocytes cell line HaCaT[J]. Journal of Third Military Medical University, 2019, 41(17): 1649-1655. DOI： 10.16016/j.1000-5404.201903210.

摘要

目的

2

建立有效的皮肤光老化HaCaT细胞模型，初步探索可能涉及的老化相关信号转导途径。

方法

2

以3

0 mJ/cm

2

中波紫外线(ultraviolet B，UVB)辐照HaCaT细胞，采用CCK-8检测细胞增殖能力，流式细胞仪分析细胞周期，衰老相关β-半乳糖苷酶(SA-β-gal)染色分析各组阳性细胞百分比，qRT-PCR和Western blot检测衰老相关蛋白p53、p21和p16的表达，细胞免疫荧光检测p21蛋白胞内定位。

结果

2

30 mJ/cm

2

UVB辐照后，HaCaT细胞增殖率均较对照组降低(

P

<

0.01)，G

2

期细胞比例较对照组增加(

P

<

0.05)，细胞SA-β-gal染色阳性率明显高于对照组(

P

<

0.01)。qRT-PCR和Western blot检测结果均显示:辐照后，HaCaT细胞中p53、p16表达明显升高，p21表达明显下降(

P

<

0.05，

P

<

0.01)。细胞免疫荧光结果显示:给予30 mJ/cm

2

UVB辐照导致p21发生明显的核移位。

结论

2

30 mJ/cm

2

UVB辐照可成功构建光老化HaCaT细胞模型，其机制可能是通过增强p21核定位及激活p16通路诱导了细胞衰老的进程。

Abstract

Objective

2

To establish an efficient skin photoaging model in HaCaT cells and investigate the possible signaling pathways of photoaging of skin.

Methods

2

After HaCaT cells were exposed to UVB at 30 mJ/cm

2

cell proliferation and cell cycle was detected by CCK-8 assay and flow cytometry. The percentages of positive cells were detected by SA-β-gal staining. The expression of p53

p21 and p16 at mRNA and protein levels were detected by qRT-PCR and Western blotting respectively. The subcellular localization of p21 was elucidated by immunofluorescence assay.

Results

2

Exposure of UVB at 30 mJ/cm

2

resulted in significantly decreased proliferation (

P

<

0.01)

increased proportion of cells at G

2

phage (

P

<

0.05)

and more positive cells to SA-β-gal (

P

<

0.05) when compared with normal control cells. The results of qRT-PCR and Western blotting showed that UVB exposure up-regulated the expression levels of p53 and p16

and down-regulated that of p21 in HaCaT cells (

P

<

0.05

P

<

0.01). Immunofluorescence assay indicated obvious nuclear localization of

p21 in the HaCaT cells after UVB exposure.

Conclusion

2

UVB-exposure at 30 mJ/cm

2

successfully establish skin photoaging model in HaCaT cells

which may be due to senescence induced by increasing nuclear localization of p21 and activating p16-dependent pathway.

关键词

Keywords

references

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