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USP14 Regulates ATF2/PIK3CD Axis to Promote Microvascular Endothelial Cell Proliferation, Migration, and Angiogenesis in Diabetic Retinopathy

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Abstract

Diabetic retinopathy (DR) is one of the leading causes of blindness in diabetic patients. However, the pathogenesis of DR is complex, and no firm conclusions have been drawn so far. It has become a hot spot in ophthalmology research to deeply study the mechanism of DR pathological changes and find effective treatment options. Human retinal microvascular endothelial cells (HRMECs) were induced by high glucose (HG) to construct DR cell model. CCK-8 assay was used to detect the viability of HRMECs. Transwell assay was used to detect the migration ability of HRMECs. Tube formation assay was used to identify the tube formation ability of HRMECs. The expressions of USP14, ATF2 and PIK3CD were detected by Western blot analysis and qRT-PCR assay. Immunoprecipitation (IP) was used to ascertain the relationship of USP14 and ATF2. To explore the regulatory relationship between ATF2 and PIK3CD by dual-luciferase reporter gene assay and Chromatin immunoprecipitation (ChIP) assay. High glucose treatment promoted the proliferation, migration, and tube formation of HRMEC, and the expressions of USP14, ATF2 and PIK3CD were significantly up-regulated. USP14 or ATF2 knockdown inhibited HG-induced HRMECs proliferation, migration, and tube formation. USP14 regulated the expression of ATF2, and ATF2 promoted PIK3CD expression. PIK3CD overexpression attenuated the inhibitory effectiveness of USP14 knockdown on proliferation, migration and tube formation of DR cell model. Here, we revealed that USP14 regulated the ATF2/PIK3CD axis to promote proliferation, migration, and tube formation in HG-induced HRMECs.

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Data Availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

DR:

Diabetic retinopathy

HG:

High glucose

HRMECs:

Human retinal microvascular endothelial cells

ChIP:

Chromatin immunoprecipitation

VEGF:

Vascular endothelial growth factor

UPS:

Ubiquitin–proteasome system

DUB:

Deubiquitinating enzyme

USP1:

Ubiquitin-specific protease 14

ATF:

Activating transcription factor 2

PI3Kδ or PIK3CD:

Phosphoinositide-3 kinase catalytic subunit δ

CCK:

Cell Counting Kit-8

FBS:

Fetal bovine serum

PBS:

Phosphate buffered saline

qRT-PCR:

Quantitative real-time PCR

PVDF:

Poly vinylidene difluoride filter

SDS-PGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

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Author notes

  1. Fu-Tao He and Xiao-Lin Fu they are the co-first authors.

Authors and Affiliations

  1. Department of Ophthalmology, Hainan West Central Hospital, No.2 Fubo East Road, Nada Town, Danzhou, 571700, Hainan Province, People’s Republic of China

    Fu-Tao He, Xiao-Lin Fu, Mo-Han Li, Chun-Yan Fu & Jian-Zhi Chen

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  1. Fu-Tao He
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  2. Xiao-Lin Fu
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  3. Mo-Han Li
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  4. Chun-Yan Fu
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Contributions

F-TH: Formal analysis; Investigation; Resources; Data Curation; X-Lf: Conceptualization; Methodology; Validation; M-HL: Writing—Original Draft; C-YF: Visualization; Supervision; J-ZC\: Writing—Review & Editing; Project administration; Funding acquisition.

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Correspondence to Xiao-Lin Fu.

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He, FT., Fu, XL., Li, MH. et al. USP14 Regulates ATF2/PIK3CD Axis to Promote Microvascular Endothelial Cell Proliferation, Migration, and Angiogenesis in Diabetic Retinopathy. Biochem Genet 61, 2076–2091 (2023). https://doi.org/10.1007/s10528-023-10358-0

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  • DOI: https://doi.org/10.1007/s10528-023-10358-0

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