Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

LncRNA GAS5 activates the HIF1A/VEGF pathway by binding to TAF15 to promote wound healing in diabetic foot ulcers

A Correction to this article was published on 05 May 2021

This article has been updated

Abstract

A diabetic foot ulcer (DFU) is one of the most devastating complications of diabetes. It has been reported that lncRNA GAS5 plays a vital role in wound healing in DFUs. However, the specific mechanism remains unclear. In this research, we aimed to investigate the role of GAS5 in wound healing in DFUs as well as the underlying mechanism. qPCR or western blotting was performed to measure the expression levels of GAS5, HIF1A, VEGF and TAF15. CCK-8 or EdU assays, flow cytometry, wound healing assays and tube formation assays were carried out to assess the proliferation, apoptosis, wound healing and in vitro angiogenesis of HUVECs, respectively. RNA pull-down and RIP assays were performed to verify the interaction between GAS5 and TAF15. ChIP and luciferase assays were conducted to verify the binding of TAF15 to the HIF1A promoter. In the DFU mouse model, H&E and Masson staining were used to determine epidermal and dermal thickness and collagen formation. GAS5 and HIF1A were downregulated in the skin tissues of DFU patients, and GAS5 overexpression promoted cell proliferation, wound healing and tubule formation in HG-treated HUVECs. In addition, GAS5 facilitated HIF1A expression by interacting with TAF15. Rescue assays demonstrated that the suppression of HIF1A/VEGF pathway activation partially reversed the functional roles of GAS5 in HUVECs. Furthermore, GAS5 accelerated wound healing by activating the HIF1A/VEGF pathway in mice with DFUs. GAS5 activates the HIF1A/VEGF pathway by binding to TAF15, resulting in accelerated wound healing in DFUs. Our findings may provide a theoretical basis for the clinical treatment of DFUs.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Fig. 1: GAS5 and HIF1A are downregulated in the skin tissues of DFU patients.
Fig. 2: HG inhibits HUVEC proliferation, wound healing and tubule formation.
Fig. 3: GAS5 overexpression promotes HG-induced HUVEC proliferation, wound healing and tubule formation.
Fig. 4: GAS5 interacts with TAF15 and promotes TAF15 expression.
Fig. 5: GAS5 accelerates HIF1A expression by interacting with TAF15.
Fig. 6: GAS5 promotes cell proliferation, wound healing and tubule formation by activating the HIF1A/VEGF pathway in HG-treated HUVECs.
Fig. 7: GAS5 accelerates DFU wound healing by activating the HIF1A/VEGF pathway.

Data availability

All data generated or analyzed during this study are included in this published article.

Change history

References

  1. 1.

    Tao F, Tang X, Tao H, Luo Y, Cao H, Xiang W, et al. Surgical treatment of diabetic foot ulcers during the COVID-19 pandemic in China. J Diabetes Complicat. 2020;34:107622.

    Article  Google Scholar 

  2. 2.

    Samad R, Yusuf S, Andriani A, Erfina E. Nurses’ perspectives on diabetic foot ulcer’s odor: a qualitative study. Enferm Clin. 2020;30:300–33.

    Article  Google Scholar 

  3. 3.

    Caruso P, Longo M, Gicchino M, Scappaticcio L, Caputo M, Maiorino MI, et al. Long-term diabetic complications as predictors of foot ulcers healing failure: a retrospective study in a tertiary-care center. Diabetes Res Clin Pract. 2020;163:108147.

    Article  Google Scholar 

  4. 4.

    Seraphim PM, Leal EC, Moura J, Gonçalves P, Gonçalves JP, Carvalho E. Lack of lymphocytes impairs macrophage polarization and angiogenesis in diabetic wound healing. Life Sci. 2020;254:117813.

    CAS  Article  Google Scholar 

  5. 5.

    Tao H, Shi P, Zhao XD, Xuan HY, Ding XS. MeCP2 inactivation of LncRNA GAS5 triggers cardiac fibroblasts activation in cardiac fibrosis. Cell Signal. 2020;74:109705.

    CAS  Article  Google Scholar 

  6. 6.

    Kumar H, Srikanth K, Park W, Lee SH, Choi BH, Kim H, et al. Transcriptome analysis to identify long non coding RNA (lncRNA) and characterize their functional role in back fat tissue of pig. Gene. 2019;703:71–82.

    CAS  Article  Google Scholar 

  7. 7.

    Li B, Luan S, Chen J, Zhou Y, Wang T, Li Z, et al. The MSC-derived exosomal lncRNA H19 promotes wound healing in diabetic foot ulcers by upregulating PTEN via microRNA-152-3p. Mol Ther Nucleic Acids. 2020;19:814–26.

    CAS  Article  Google Scholar 

  8. 8.

    Liu XQ, Duan LS, Chen YQ, Jin XJ, Zhu NN, Zhou X, et al. lncRNA MALAT1 accelerates wound healing of diabetic mice transfused with modified autologous blood via the HIF-1α signaling pathway. Mol Ther Nucleic Acids. 2019;17:504–15.

    CAS  Article  Google Scholar 

  9. 9.

    Carter G, Miladinovic B, Patel AA, Deland L, Mastorides S, Patel NA. Circulating long noncoding RNA GAS5 levels are correlated to prevalence of type 2 diabetes mellitus. BBA Clin. 2015;4:102–7.

    Article  Google Scholar 

  10. 10.

    Shi Y, Parag S, Patel R, Lui A, Murr M, Cai J, et al. Stabilization of lncRNA GAS5 by a small molecule and its implications in diabetic adipocytes. Cell Chem Biol. 2019;26:319–330.e6.

    CAS  Article  Google Scholar 

  11. 11.

    Sawaya AP, Pastar I, Stojadinovic O, Lazovic S, Davis SC, Gil J, et al. Topical mevastatin promotes wound healing by inhibiting the transcription factor c-Myc via the glucocorticoid receptor and the long non-coding RNA Gas5. J Biol Chem. 2018;293:1439–49.

    CAS  Article  Google Scholar 

  12. 12.

    Ruan X, Zheng J, Liu X, Liu Y, Liu L, Ma J, et al. lncRNA LINC00665 stabilized by TAF15 impeded the malignant biological behaviors of glioma cells via STAU1-mediated mRNA degradation. Mol Ther Nucleic Acids. 2020;20:823–40.

    CAS  Article  Google Scholar 

  13. 13.

    Ballarino M, Jobert L, Dembele D, de la Grange P, Auboeuf D, Tora L. TAF15 is important for cellular proliferation and regulates the expression of a subset of cell cycle genes through miRNAs. Oncogene. 2013;32:4646–55.

    CAS  Article  Google Scholar 

  14. 14.

    Pan L, Li Y, Jin L, Li J, Xu A. TRPM2-AS promotes cancer cell proliferation through control of TAF15. Int J Biochem Cell Biol. 2020;120:105683.

    CAS  Article  Google Scholar 

  15. 15.

    Chen L, Chen Q, Kuang S, Zhao C, Yang L, Zhang Y, et al. USF1-induced upregulation of LINC01048 promotes cell proliferation and apoptosis in cutaneous squamous cell carcinoma by binding to TAF15 to transcriptionally activate YAP1. Cell Death Dis. 2019;10:296.

    CAS  Article  Google Scholar 

  16. 16.

    Jiang YZ, Li Y, Wang K, Dai CF, Huang SA, Chen DB, et al. Distinct roles of HIF1A in endothelial adaptations to physiological and ambient oxygen. Mol Cell Endocrinol. 2014;391:60–67.

    CAS  Article  Google Scholar 

  17. 17.

    Li L, Wang M, Mei Z, Cao W, Yang Y, Wang Y, et al. lncRNAs HIF1A-AS2 facilitates the up-regulation of HIF-1α by sponging to miR-153-3p, whereby promoting angiogenesis in HUVECs in hypoxia. Biomed Pharmacother. 2017;96:165–72.

    CAS  Article  Google Scholar 

  18. 18.

    Dong SM, Cui JH, Zhang W, Zhang XW, Kou TC, Cai QC, et al. Inhibition of translation initiation factor eIF4A is required for apoptosis mediated by Microplitis bicoloratus bracovirus. Arch Insect Biochem Physiol. 2017;96:e21423.

    Article  Google Scholar 

  19. 19.

    Cui JH, Dong SM, Chen CX, Xiao W, Cai QC, Zhang LD, et al. Microplitis bicoloratus bracovirus modulates innate immune suppression through the eIF4E–eIF4A axis in the insect Spodoptera litura. Dev Comp Immunol. 2019;95:101–7.

    CAS  Article  Google Scholar 

  20. 20.

    Liu XH, Sun M, Nie FQ, Ge YB, Zhang EB, Yin DD, et al. Lnc RNA HOTAIR functions as a competing endogenous RNA to regulate HER2 expression by sponging miR-331-3p in gastric cancer. Mol Cancer. 2014;13:92.

    CAS  Article  Google Scholar 

  21. 21.

    He ZY, Wei TH, Zhang PH, Zhou J, Huang XY. Long noncoding RNA-antisense noncoding RNA in the INK4 locus accelerates wound healing in diabetes by promoting lymphangiogenesis via regulating miR-181a/Prox1 axis. J Cell Physiol. 2019;234:4627–40.

    CAS  Article  Google Scholar 

  22. 22.

    Li J, Li SX, Gao XH, Zhao LF, Du J, Wang TY, et al. HIF1A and VEGF regulate each other by competing endogenous RNA mechanism and involve in the pathogenesis of peritoneal fibrosis. Pathol Res Pract. 2019;215:644–52.

    CAS  Article  Google Scholar 

  23. 23.

    Hung SY, Tsai JS, Yeh JT, Chen KH, Lin CN, Yang HM, et al. Amino acids and wound healing in people with limb-threatening diabetic foot ulcers. J Diabetes Complicat. 2019;33:107403.

    Article  Google Scholar 

  24. 24.

    Kiya K, Kubo T. Neurovascular interactions in skin wound healing. Neurochem Int. 2019;125:144–50.

    CAS  Article  Google Scholar 

  25. 25.

    Zou J, Liu KC, Wang WP, Xu Y. Circular RNA COL1A2 promotes angiogenesis via regulating miR-29b/VEGF axis in diabetic retinopathy. Life Sci. 2020;256:117888.

    CAS  Article  Google Scholar 

  26. 26.

    Zeng B, Li Y, Jiang F, Wei C, Chen G, Zhang W, et al. LncRNA GAS5 suppresses proliferation, migration, invasion, and epithelial-mesenchymal transition in oral squamous cell carcinoma by regulating the miR-21/PTEN axis. Exp Cell Res. 2019;374:365–73.

    CAS  Article  Google Scholar 

  27. 27.

    Apaya MK, Kuo TF, Yang MT, Yang G, Hsiao CL, Chang SB, et al. Phytochemicals as modulators of β-cells and immunity for the therapy of type 1 diabetes: Recent discoveries in pharmacological mechanisms and clinical potential. Pharmacol Res. 2020;156:104754.

    CAS  Article  Google Scholar 

  28. 28.

    Liu GM, Zeng HD, Zhang CY, Xu JW. Key genes associated with diabetes mellitus and hepatocellular carcinoma. Pathol Res Pract. 2019;215:152510.

    CAS  Article  Google Scholar 

  29. 29.

    Fadini GP, Menegazzo L, Rigato M, Scattolini V, Poncina N, Bruttocao A, et al. NETosis Delays diabetic wound healing in mice and humans. Diabetes. 2016;65:1061–71.

    CAS  Article  Google Scholar 

  30. 30.

    Zhang R, Han X, Huang T, Wang X. Danggui buxue tang inhibited mesangial cell proliferation and extracellular matrix accumulation through GAS5/NF-kappaB pathway. Biosci Rep. 2019;39:BSR20181740.

    CAS  Article  Google Scholar 

  31. 31.

    Xie C, Wu W, Tang A, Luo N, Tan Y. lncRNA GAS5/miR-452-5p reduces oxidative stress and pyroptosis of high-glucose-stimulated renal tubular cells. Diabetes Metab Syndr Obes. 2019;12:2609–17.

    CAS  Article  Google Scholar 

  32. 32.

    Ruan X, Zheng J, Liu X, Liu Y, Liu L, Ma J, et al. lncRNA LINC00665 stabilized by TAF15 impeded the malignant biological behaviors of glioma cells via STAU1-mediated mRNA degradation. Mol Ther Nucleic Acids. 2020;20:823–40.

    CAS  Article  Google Scholar 

  33. 33.

    Wang R, Ma Z, Feng L, Yang Y, Tan C, Shi Q, et al. LncRNA MIR31HG targets HIF1A and P21 to facilitate head and neck cancer cell proliferation and tumorigenesis by promoting cell-cycle progression. Mol Cancer. 2018;17:162–7.

    CAS  Article  Google Scholar 

Download references

Funding

This work was supported by General Program of National Natural Science Foundation of China (81870573).

Author information

Affiliations

Authors

Contributions

Guarantor of integrity of the entire study: G-DX. Study concepts and design: G-DX and W-XP. Definition of intellectual content: P-XH and TZ. Literature research: TZ and Y-QZ. Clinical studies: W-XP, TZ, Y-QZ, KP, LX and J-JY. Experimental studies and manuscript editing: W-XP and TZ. Data acquisition: W-XP and Y-QZ. Data analysis: L-JL and KP. Statistical analysis: LX and J-JY. Manuscript preparation: W-XP and P-XH. Manuscript review: P-XH and G-DX.

Corresponding author

Correspondence to Guang-Da Xiang.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Ethics approval

Experiment protocols were approved by the Ethics Committee of Yiyang Central Hospital and experimental procedures were conducted according to the Declaration of Helsinki Principles. The animal experiments involved in this paper were carried out in accordance with the Guidelines for the Care and Use of Laboratory Animals issued by the National Institutes of Health.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

The original online version of this article was revised: Due to a typeset error the Summary was omitted.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Peng, WX., He, PX., Liu, LJ. et al. LncRNA GAS5 activates the HIF1A/VEGF pathway by binding to TAF15 to promote wound healing in diabetic foot ulcers. Lab Invest (2021). https://doi.org/10.1038/s41374-021-00598-2

Download citation

Search

Quick links