Research papercircRNA_0084043 contributes to the progression of diabetic retinopathy via sponging miR-140-3p and inducing TGFA gene expression in retinal pigment epithelial cells
Introduction
Diabetes mellitus is a chronic disease with great morbidity and mortality across the global (Santos et al., 2017). Diabetic retinopathy (DR) is a serious microvascular complication of diabetes all over the world (Murchison et al., 2017). As well established, many metabolic disorders are closely associated with the pathogenesis of DR (Arroba et al., 2018). Currently, the molecular mechanisms of DR progression are still poorly known and no effective treatments have been developed for DR since now. Our current research was aimed to identify risk factors for DR and indicate a novel target.
Circular RNA is abundantly expressed in eukaryotes and can exert important biological roles (Kristensen et al., 2018, Rybak-Wolf et al., 2015). Increasing evidence has shown that circRNA can function as a ceRNA via sponging miRNAs (Hansen et al., 2013, Zhong et al., 2018). Clinical data has displayed that circRNAs are involved in a variety of diseases, which suggest that circRNAs has a wide effect on diseases (Zhou et al., 2018, Zhuo et al., 2020, Li et al., 2020). For example, circ-CMPK1 can contribute to lung cancer cell proliferation through sponging miR-302e (Cui et al., 2019). CircRNA ITCH can repress the progression of prostate cancer through inducing HOXB13 and sponging miR-17-5p (Wang et al., 2019). Recently, the clinical significance of circRNAs has been shown in DR (Zhang et al., 2017, Gu et al., 2017). For instance, loss of circRNA DMNT3B results in diabetic retinal vascular dysfunction via regulating miR-20b-5p and BAMBI (Zhu et al., 2019). However, the biological effect of circRNA_0084043 on DR progression remains barley known.
Currently, we demonstrated that circRNA_0084043 promoted cell proliferation and inhibited cell apoptosis induced by HG. Meanwhile, oxidative stress and inflammation were also repressed by loss of circRNA_0084043. Then, we further investigated the interaction between circRNA_0084043 and miR-140-3p, since miR-140-3p was predicted as a target of circRNA_0084043. Moreover, inhibitors of miR-140-3p successfully reversed the function of circRNA_0084043 siRNA on DR progression via targeting TGFA.
Section snippets
Cell culture
ARPE-19 was obtained from ATCC (Manassas, VA, USA). Cells were incubated in DMEM with 10% FBS (Sigma, St. Louis, MO, USA), 1% non-essential amino acids, 100 g/ml streptomycin (Invitrogen, Carlsbad, CA, USA) and 100U/ml penicillin (Invitrogen, Carlsbad, CA, USA) in 5% CO2 at 37 °C.
Cell transfection
miR-140-3p mimics (10 nM), inhibitors (100 nM), circRNA_0084043 (50 nM) or the corresponding negative controls were obtained from GenePharma (Shanghai, China). Transfection was carried out by Lipofectamine 3000
circRNA_0084043 was significantly elevated in DR cell model
Firstly, we established cell injury model of DR. circRNA_0084043 was tested in ARPE-19 cells treated with HG. The data as shown in Fig. 1A denoted that circRNA_0084043 was remarkably increased by HG in vitro.
Down-regulated circRNA_0084043 protected ARPE-19 cell against HG-inhibited cell proliferation and HG-induced cell apoptosis
Moreover, in order to study the role of circRNA_0084043 on HG-induced cell injury, ARPE-19 cells were infected with circRNA_0084043 siRNA. As shown in Fig. 2A, circRNA_0084043 was obviously increased by circRNA_0084043 siRNA in ARPE-19 cells. Results in Fig. 2B exhibited that cell viability
Discussion
Recently, circRNAs have obtained increasing attention due to their roles in gene modulation of various human diseases (Barrett and Salzman, 2016). Diabetic vascular complications remain the leading causes of blindness (Tabit et al., 2010, Sena et al., 2013). Meanwhile, diabetes-triggered retinal vascular dysfunction is a main cause of blindness (Stitt et al., 2016). In our study, we observed that circRNA_0084043 was elevated in ARPE-19 cells incubated with HG. In addition, we found that loss of
Data Availability Statement
The data are available upon reasonable request.
Funding
This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.
CRediT authorship contribution statement
Ying Li: Writing - original draft, Methodology, Data curation. Ting Chen: Software, Methodology, Supervision, Validation, Visualization. Chengliang Wan: Investigation, Writing - original draft, Data curation, Formal analysis, Software, Methodology, Supervision, Validation, Visualization. Hongyan Cang: Conceptualization, Project administration, Writing - review & editing, Resources.
Declaration of Competing Interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgements
None.
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These authors contributed equally to this work.