Silencing LncRNA PVT1 Reverses High Glucose-Induced Regulation of the High Expression of PVT1 in HRMECs by Targeting miR-128-3p

Abstract

This paper aims to discuss the possibility of lncRNA PVT1 as a diagnostic biomarker for diabetic retinopathy (DR) and explore the underlying mechanism. Real-time quantitative polymerase chain reaction (RT-qPCR) was selected to determine the expression level of lncRNA PVT1 in the serum of all subjects. The receiver operating characteristic (ROC) curve reflected the diagnostic significance of PVT1 for DR patients. The Cell Counting Kit-8 (CCK-8) and Transwell assays were used to evaluate the effect of PVT1 expression on the proliferation and migration of human retinal microvascular endothelial cells (HRMECs). The luciferase reporter gene was selected to verify the interaction between PVT1 and miR-128-3p. The relative expression level of PVT1 in serum was higher in both the DB and DR group than in the healthy controls group (HC), and it was highest in the DR group. ROC curve indicated that serum PVT1 could distinguish between HC and DB patients, DB patients and DR patients, respectively. In vitro, high glucose induction significantly increased the proliferation and migration capabilities of HRMECs, but silencing PVT1 (si-PVT1) downregulated the proliferation and migration capabilities of HRMECs. The detection of luciferase reporter gene showed that lncRNA PVT1 targeted miR-128-3p, and there was a negative correlation in the serum of DR patients. In conclusion, this study confirmed that lncRNA PVT1 might regulate the process of DR by targeting miR-128-3p, and has the potential as a biomarker for the diagnosis of DR.

Key words

diabetic retinopathy - lncRNA PVT1 - miR-128-3p - diagnosis - high glucose

Publication History

Received: 17 November 2021

Accepted after revision: 21 December 2021

Article published online:
07 February 2022

• References

• 1 Kerner W, Brückel J. Definition, classification and diagnosis of diabetes mellitus. Exp Clin Endocrinol Diabetes 2014; 122: 384-386
• 2 Schmidt AM. Highlighting diabetes mellitus: the epidemic continues. Arterioscler Thromb Vasc Biol 2018; 38: e1-e8
• 3 Harris-Hayes M, Schootman M, Schootman JC. et al. The role of physical therapists in fighting the type 2 diabetes epidemic. J Orthop Sports Phys Ther 2020; 50: 5-16
• 4 Feng W, Wang Y, Guo N. et al. Effects of astaxanthin on inflammation and insulin resistance in a mouse model of gestational diabetes mellitus. Dose Response. 2020 18. 1559325820926765
• 5 Piko P, Werissa NA, Fiatal S. et al. Impact of genetic factors on the age of onset for type 2 diabetes mellitus in addition to the conventional risk factors. J Pers Med 2020; 11: 6
• 6 Nam JS, Han JW, Lee SB. et al. Calpain-10 and adiponectin gene polymorphisms in Korean type 2 diabetes patients. Endocrinol Metab (Seoul) 2018; 33: 364-371
• 7 Asare-Bediako B, Noothi SK, Li Calzi S. et al. Characterizing the retinal phenotype in the high-fat diet and western diet mouse models of prediabetes. Cells 2020; 9: 464
• 8 Zhang B, Wang Q, Zhang X. et al. Association between self-care agency and depression and anxiety in patients with diabetic retinopathy. BMC Ophthalmol 2021; 21: 123
• 9 Mao XB, Cheng YH, Peng KS. et al. Sirtuin (Sirt) 3 overexpression prevents retinopathy in streptozotocin-induced diabetic rats. Med Sci Monit 2020; 26: e920883
• 10 Zeng M, Shen J, Liu Y. et al. The HIF-1 antagonist acriflavine: visualization in retina and suppression of ocular neovascularization. J Mol Med (Berl) 2017; 95: 417-429
• 11 Yang X, Cao J, Du Y. et al. Angiopoietin-like protein 4 (ANGPTL4) induces retinal pigment epithelial barrier breakdown by activating signal transducer and activator of transcription 3 (STAT3): evidence from ARPE-19 cells under hypoxic condition and diabetic rats. Med Sci Monit 2019; 25: 6742-6754
• 12 Cheung N, Mitchell P, Wong TY. Diabetic retinopathy. Lancet 2010; 376: 124-136
• 13 Li Z, Lü M, Zhou Y. et al. Role of long non-coding RNAs in the chemoresistance of gastric cancer: a systematic review. Onco Targets Ther 2021; 14: 503-518
• 14 Yuan F, Lou Z, Zhou Z. et al. Long non‑coding RNA KCNQ1OT1 promotes nasopharyngeal carcinoma cell cisplatin resistance via the miR‑454/USP47 axis. Int J Mol Med 2021; 47: 54
• 15 Thomas AA, Biswas S, Feng B. et al. lncRNA H19 prevents endothelial-mesenchymal transition in diabetic retinopathy. Diabetologia 2019; 62: 517-530
• 16 Sun Y, Liu YX. LncRNA HOTTIP improves diabetic retinopathy by regulating the p38-MAPK pathway. Eur Rev Med Pharmacol Sci 2018; 22: 2941-2948
• 17 Yang J, Zhao S, Tian F. SP1-mediated lncRNA PVT1 modulates the proliferation and apoptosis of lens epithelial cells in diabetic cataract via miR-214-3p/MMP2 axis. J Cell Mol Med 2020; 24: 554-561
• 18 Cheng Y, Hu Q, Zhou J. Silencing of lncRNA PVT1 ameliorates streptozotocin-induced pancreatic b cell injury and enhances insulin secretory capacity by regulating miR-181a-5p. Can J Physiol Pharmacol 2021; 99: 303-312
• 19 Pang B, Li QW, Qin YL. et al. Traditional chinese medicine for diabetic retinopathy: a systematic review and meta-analysis. Medicine (Baltimore) 2020; 99: e19102
• 20 Zeng Q, Liu J. Silencing circ_0001879 inhibits the proliferation and migration of human retinal microvascular endothelial cells under high-glucose conditions via modulating miR-30-3p. Gene 2020; 760: 144992
• 21 Xu Y, Zhang Y, Liang H. et al. Coumestrol mitigates retinal cell inflammation, apoptosis, and oxidative stress in a rat model of diabetic retinopathy via activation of SIRT1. Aging (Albany NY) 2021; 13: 5342-5357
• 22 Ehrke-Schulz E, Bergmann T, Schiwon M. et al. Quantification of designer nuclease induced mutation rates: a direct comparison of different methods. Mol Ther Methods Clin Dev 2016; 3: 16047
• 23 Liu Y, Gao M, An J. et al. Dysregulation of MiR-30a-3p/gastrin enhances tumor growth and invasion throughSTAT3/MMP11 pathway in gastric cancer. Onco Targets Ther 2020; 13: 8475-8493
• 24 Zou H, Wu LX, Yang Y. et al. lncRNAs PVT1 and HAR1A are prognosis biomarkers and indicate therapy outcome for diffuse glioma patients. Oncotarget 2017; 8: 78767-78780
• 25 Zhang Y, Mo WJ, Wang X. et al. Microarray‑based bioinformatics analysis of the prospective target gene network of key miRNAs influenced by long non‑coding RNA PVT1 in HCC. Oncol Rep 2018; 40: 226-240
• 26 Zhou C, Yi C, Yi Y. et al. LncRNA PVT1 promotes gemcitabine resistance of pancreatic cancer via activating Wnt/b-catenin and autophagy pathway through modulating the miR-619-5p/Pygo2 and miR-619-5p/ATG14 axes. Mol Cancer 2020; 19: 118
• 27 Zhao W, Hong L, Wang J. et al. Cyclizing-berberine A35 induces G2/M arrest and apoptosis by activating YAP phosphorylation (Ser127). J Exp Clin Cancer Res 2018; 37: 98
• 28 Shigeyasu K, Toden S, Ozawa T. et al. The PVT1 lncRNA is a novel epigenetic enhancer of MYC, and a promising risk-stratification biomarker in colorectal cancer. Mol Cancer 2020; 19: 155
• 29 Wang Y, Chen W, Lian J. et al. The lncRNA PVT1 regulates nasopharyngeal carcinoma cell proliferation via activating the KAT2A acetyltransferase and stabilizing HIF-1a. Cell Death Differ 2020; 27: 695-710
• 30 Liu DW, Zhang JH, Liu FX. et al. Silencing of long noncoding RNA PVT1 inhibits podocyte damage and apoptosis in diabetic nephropathy by upregulating FOXA1. Exp Mol Med 2019; 51: 1-15
• 31 Wang X, Wang G, Zhang L. et al. LncRNA PVT1 promotes the growth of HPV positive and negative cervical squamous cell carcinoma by inhibiting TGF-β1. Cancer Cell Int 2018; 18: 70
• 32 Zhu M, Lv Q, Huang H. et al. Identification of a four-long non-coding RNA signature in predicting breast cancer survival. Oncol Lett 2020; 19: 221-228
• 33 Lu D, Luo P, Wang Q. et al. lncRNA PVT1 in cancer: a review and meta-analysis. Clin Chim Acta 2017; 474: 1-7
• 34 Xiao F, Li L, Fu JS. et al. Regulation of the miR-19b-mediated SOCS6-JAK2/STAT3 pathway by lncRNA MEG3 is involved in high glucose-induced apoptosis in hRMECs. Biosci Rep 2020; 40: BSR20194370
• 35 Yu L, Fu J, Yu N. et al. Long noncoding RNA MALAT1 participates in the pathological angiogenesis of diabetic retinopathy in an oxygen-induced retinopathy mouse model by sponging miR-203a-3p. Can J Physiol Pharmacol 2020; 98: 219-227
• 36 Liu K, Xu Q. LncRNA PVT1 regulates gallbladder cancer progression through miR-30d-5p. J Biol Regul Homeost Agents 2020; 34: 875-883
• 37 Guo J, Xiao F, Ren W. et al. Circular ribonucleic acid circFTO promotes angiogenesis and impairs blood-retinal barrier via targeting the miR-128-3p/thioredoxin interacting protein axis in diabetic retinopathy. Front Mol Biosci 2021; 8: 685466