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On-line Access: 2022-03-09

Received: 2021-02-26

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Zhiyong WANG


Xiqiao WANG


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Journal of Zhejiang University SCIENCE B

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Transition of autophagy and apoptosis in fibroblasts depends on dominant expression of HIF-1α or p53

Author(s):  Min LI, Yidan SU, Xiaoyuan GAO, Jiarong YU, Zhiyong WANG, Xiqiao WANG

Affiliation(s):  Department of Burn, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China; more

Corresponding email(s):  wxqiao2002@hotmail.com, wzy10830@rjh.com.cn

Key Words:  Hypertrophic scar; Hypoxia-inducible factor-1α (HIF-1α); p53; Autophagy; Apoptosis

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Min LI, Yidan SU, Xiaoyuan GAO, Jiarong YU, Zhiyong WANG, Xiqiao WANG. Transition of autophagy and apoptosis in fibroblasts depends on dominant expression of HIF-1α or p53[J]. Journal of Zhejiang University Science B, 2022, 23(3): 204-217.

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author="Min LI, Yidan SU, Xiaoyuan GAO, Jiarong YU, Zhiyong WANG, Xiqiao WANG",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Transition of autophagy and apoptosis in fibroblasts depends on dominant expression of HIF-1α or p53
%A Min LI
%A Yidan SU
%A Xiaoyuan GAO
%A Jiarong YU
%A Zhiyong WANG
%A Xiqiao WANG
%J Journal of Zhejiang University SCIENCE B
%V 23
%N 3
%P 204-217
%@ 1673-1581
%D 2022
%I Zhejiang University Press & Springer

T1 - Transition of autophagy and apoptosis in fibroblasts depends on dominant expression of HIF-1α or p53
A1 - Min LI
A1 - Yidan SU
A1 - Xiaoyuan GAO
A1 - Jiarong YU
A1 - Zhiyong WANG
A1 - Xiqiao WANG
J0 - Journal of Zhejiang University Science B
VL - 23
IS - 3
SP - 204
EP - 217
%@ 1673-1581
Y1 - 2022
PB - Zhejiang University Press & Springer
ER -

It has been revealed that hypoxia is dynamic in hypertrophic scars; therefore, we considered that it may have different effects on hypoxia-inducible factor-1α (HIF-1α) and p53 expression. Herein, we aimed to confirm the presence of a teeterboard-like conversion between HIF-1α and p53, which is correlated with scar formation and regression. Thus, we obtained samples of normal skin and hypertrophic scars to identify the differences in HIF-1α and autophagy using immunohistochemistry and transmission electron microscopy. In addition, we used moderate hypoxia in vitro to simulate the proliferative scar, and silenced HIF-1α or p53 gene expression or triggered overexpression to investigate the changes of HIF-1α and p53 expression, autophagy, apoptosis, and cell proliferation under this condition. HIF-1α, p53, and autophagy-related proteins were assayed using western blotting and immunofluorescence, whereas apoptosis was detected using flow cytometry analysis, and cell proliferation was detected using cell counting kit-8 (CCK-8) and 5-bromo-2'-deoxyuridine (BrdU) staining. Furthermore, immunoprecipitation was performed to verify the binding of HIF-1α and p53 to transcription cofactor p300. Our results demonstrated that, in scar tissue, HIF-1α expression increased in parallel with autophagosome formation. Under hypoxia, HIF-1α expression and autophagy were upregulated, whereas p53 expression and apoptosis were downregulated in vitro. HIF-1α knockdown downregulated autophagy, proliferation, and p300-bound HIF-1α, and upregulated p53 expression, apoptosis, and p300-bound p53. Meanwhile, p53 knockdown induced the opposite effects and enhanced HIF-1α, whereas p53 overexpression resulted in the same effects and reduced HIF-1α. Our results suggest a teeterboard-like conversion between HIF-1α and p53, which is linked with scar hyperplasia and regression.


方法:首先,通过免疫组织化学和透射电镜观察增生期增生性瘢痕相对于正常皮肤中HIF-1α的表达与自噬的变化;其次,在体外模拟的中度缺氧环境中培养人皮肤成纤维细胞(HDFs),通过western blot和免疫荧光染色检测细胞自噬相关蛋白,利用流式细胞技术检测细胞凋亡率,采用细胞计数试剂盒8(CCK-8)和5-溴脱氧尿嘧啶核苷(BrdU)染色检测细胞增殖活力;再则使用慢病毒载体转染HDFs,分别敲低HIF-1α、敲低p53、过表达p53,检测基因表达规律及相应细胞自噬和凋亡等变化;最后,应用免疫沉淀验证HIF-1α和p53竞争性结合p300。


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