Abstract
Background: Diabetic retinopathy (DR) is currently the leading cause of blindness and visual disability in adults with diabetes mellitus (DM). Neovascularization has been identified as an important clinical property in DR, however, the exact mechanisms in DR neovascularization are still unclear and need further elucidation.Methods: Quantitative real-time PCR (qRT-PCR) was conducted to detect the expression level of long non-coding RNA (lncRNA)-metastasis associated lung adenocarcinoma transcript 1 (MALAT1), miR-125b and vascular endothelial-cadherin (VE-cadherin) in human retina microvascular endothelial cells (hRMECs) treated with high glucose (HG). Luciferase assay was used to detect interaction of MALAT1 with miR-125b and miR-125b with VE-cadherin. MTT assay, transwell assay, tube formation assay and vascular permeability assay were conducted to detect the cell viability, migration tube formation ability and permeability of hRMECs, respectively. ELISA was used to examine the release of VE-cadherin and vascular endothelial growth factor (VEGF). Western blotting was used to access the protein expression of VE-cadherin, VEGF, β-catenin, matrix metalloproteinase (MMP) 2 (MMP2) and MMP9.Results: MALAT1 and VE-cadherin were up-regulated while miR-125b was down-regulated in hRMECs treated with HG. MALAT1 could competitively bind to miR-125b against VE-cadherin at the site of 3′-untranslated region (3′-UTR), leading to the up-regulation of VE-cadherin. Knockdown of MALAT1 inhibited the proliferation, migration, tube formation and vascular permeability of hRMECs induced by HG through up-regulating miR-125b. Furthermore, we found the deletion of MALAT1 suppressed the VE-cadherin/β-catenin complex and neovascularization related proteins expression, which was up-regulated by HG.Conclusion: Knockdown of MALAT1 inhibited cell proliferation, migration and angiogenesis of hRMECs via suppressing the VE-cadherin/β-catenin complex through targeting miR-125b. Inhibition of MALAT1 may serve as a potential target for anti-angiogenic therapy for DR.
Highlights
Diabetic retinopathy (DR) is a frequent microvascular complication of patients with diabetes mellitus (DM) and one of the most common sight-threatening conditions, eventually leading to severe vitreous cavity bleeding, retinal detachment and vision loss [1]
To explore the effects of high glucose (HG) on the expression of metastasis associated lung adenocarcinoma transcript 1 (MALAT1) and miR-125b, we used Quantitative real-time PCR (qRT-PCR) and Western blotting in human retina microvascular endothelial cell (hRMEC) exposure of HG (25 mM). qRT-PCR analysis revealed that the levels of MALAT1 and VE-cadherin were significantly increased after HG treatment (Figure 1A,C), while the level of miR-125b declined sharply (Figure 1B)
DR is a chronic and serious eye complication associated with DM, and people with DR present an increased risk of other microvascular and macrovascular complications associated with diabetes
Summary
Diabetic retinopathy (DR) is a frequent microvascular complication of patients with diabetes mellitus (DM) and one of the most common sight-threatening conditions, eventually leading to severe vitreous cavity bleeding, retinal detachment and vision loss [1]. The severity of diabetes plays a key role in DR, and several genes have been reported to be relevant with DR neovascularization [4] Among these markers, vascular endothelial-cadherin (VE-cadherin) stands out as a reliable biomarker of angiogenesis activity [5]. High glucose (HG) could increase VE-cadherin expression in human retina microvascular endothelial cells (hRMECs) [8]. Methods: Quantitative real-time PCR (qRT-PCR) was conducted to detect the expression level of long non-coding RNA (lncRNA)-metastasis associated lung adenocarcinoma transcript 1 (MALAT1), miR-125b and vascular endothelial-cadherin (VE-cadherin) in human retina microvascular endothelial cells (hRMECs) treated with high glucose (HG). Conclusion: Knockdown of MALAT1 inhibited cell proliferation, migration and angiogenesis of hRMECs via suppressing the VE-cadherin/β-catenin complex through targeting miR-125b. Inhibition of MALAT1 may serve as a potential target for anti-angiogenic therapy for DR
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