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Abstract：
OBJECTIVE: Diabetic retinopathy (DR) is a common complication of diabetes, and recent findings have shown that long noncoding RNAs (lncRNAs) may be involved in its pathogenesis. Through bioinformatics analysis, we found that lncRNA ATP2B2-IT2 may be involved in this process. This study primarily investigated the expression of the lncRNA ATP2B2-IT2 in human retinal microvascular endothelial cells (HRMECs) under high-glucose conditions and its effects on HRMEC proliferation, migration, and neovascularization.
METHODS: We used RT‒PCR to assess the expression levels of lncRNA ATP2B2-IT2 and vascular endothelial growth factor (VEGF) in HRMECs under normal glucose (5.5 mmol/L) and high glucose (30 mmol/L) conditions. HRMECs were subsequently divided into four groups: the normal glucose (NG), high glucose (HG), high glucose with lncRNA ATP2B2-IT2 silencing (HG + si-lncRNA ATP2B2-IT2), and high glucose with silencing control (HG + si-NC) groups. The expression levels of the lncRNA ATP2B2-IT2 and VEGF in each group were determined using RT‒PCR. Thereafter, cell proliferation, migration, and neovascularization were assessed using CCK-8, Transwell, and tube formation assays, respectively.
RESULTS: RT‒PCR revealed that the expression levels of the lncRNA ATP2B2-IT2 and VEGF were greater in the HG group than in the NG group (P < 0.05). After silencing of the lncRNA ATP2B2-IT2, the expression of VEGF decreased significantly (P < 0.05). Subsequent CCK-8, Transwell, and tube formation assays demonstrated that compared to those in the NG group, the HRMECs in the HG group exhibited significantly increased proliferation, migration, and neovascularization (P < 0.05). However, after silencing of the lncRNA ATP2B2-IT2, the proliferation, migration, and neovascularization of HRMECs were significantly decreased in the HG + si-lncRNA ATP2B2-IT2 group compared to those in the HG group (P < 0.05).
CONCLUSIONS: LncRNA ATP2B2-IT2 may promote the proliferation, migration and neovascularization of HRMECs under high-glucose conditions.
METHODS: We used RT‒PCR to assess the expression levels of lncRNA ATP2B2-IT2 and vascular endothelial growth factor (VEGF) in HRMECs under normal glucose (5.5 mmol/L) and high glucose (30 mmol/L) conditions. HRMECs were subsequently divided into four groups: the normal glucose (NG), high glucose (HG), high glucose with lncRNA ATP2B2-IT2 silencing (HG + si-lncRNA ATP2B2-IT2), and high glucose with silencing control (HG + si-NC) groups. The expression levels of the lncRNA ATP2B2-IT2 and VEGF in each group were determined using RT‒PCR. Thereafter, cell proliferation, migration, and neovascularization were assessed using CCK-8, Transwell, and tube formation assays, respectively.
RESULTS: RT‒PCR revealed that the expression levels of the lncRNA ATP2B2-IT2 and VEGF were greater in the HG group than in the NG group (P < 0.05). After silencing of the lncRNA ATP2B2-IT2, the expression of VEGF decreased significantly (P < 0.05). Subsequent CCK-8, Transwell, and tube formation assays demonstrated that compared to those in the NG group, the HRMECs in the HG group exhibited significantly increased proliferation, migration, and neovascularization (P < 0.05). However, after silencing of the lncRNA ATP2B2-IT2, the proliferation, migration, and neovascularization of HRMECs were significantly decreased in the HG + si-lncRNA ATP2B2-IT2 group compared to those in the HG group (P < 0.05).
CONCLUSIONS: LncRNA ATP2B2-IT2 may promote the proliferation, migration and neovascularization of HRMECs under high-glucose conditions.
摘要：
目的：糖尿病视网膜病变（DR）是糖尿病的常见并发症，和最近的研究结果表明，长链非编码RNA（lncRNAs）可能参与其发病机制。通过生物信息学分析,我们发现lncRNAATP2B2-IT2可能参与了这一过程。这项研究主要研究了高糖条件下lncRNAATP2B2-IT2在人视网膜微血管内皮细胞（HRMEC）中的表达及其对HRMEC增殖的影响。迁移,和新血管形成。
方法：我们使用RT-PCR评估了正常葡萄糖（5.5mmol/L）和高糖（30mmol/L）条件下HRMEC中lncRNAATP2B2-IT2和血管内皮生长因子（VEGF）的表达水平。随后将HRMEC分为四组：正常葡萄糖（NG），高葡萄糖（HG），高葡萄糖与lncRNAATP2B2-IT2沉默（HGsi-lncRNAATP2B2-IT2），和高糖沉默对照组(HG+si-NC)。使用RT-PCR测定各组中lncRNAATP2B2-IT2和VEGF的表达水平。此后,细胞增殖，迁移,使用CCK-8，Transwell，和试管形成测定，分别。
结果：RT-PCR显示，HG组lncRNAATP2B2-IT2和VEGF的表达水平高于NG组（P<0.05）。lncRNAATP2B2-IT2沉默后,VEGF的表达显著降低(P<0.05)。随后的CCK-8,Transwell,和试管形成试验表明，与NG组相比，HG组的HRMEC表现出显著增加的增殖，迁移,新生血管形成(P<0.05)。然而,lncRNAATP2B2-IT2沉默后，增殖，迁移,与HG组相比，HGsi-lncRNAATP2B2-IT2组的HRMECs新生血管明显减少（P<0.05）。
结论：LncRNAATP2B2-IT2可能促进细胞增殖，高葡萄糖条件下HRMEC的迁移和新生血管形成。
方法：我们使用RT-PCR评估了正常葡萄糖（5.5mmol/L）和高糖（30mmol/L）条件下HRMEC中lncRNAATP2B2-IT2和血管内皮生长因子（VEGF）的表达水平。随后将HRMEC分为四组：正常葡萄糖（NG），高葡萄糖（HG），高葡萄糖与lncRNAATP2B2-IT2沉默（HGsi-lncRNAATP2B2-IT2），和高糖沉默对照组(HG+si-NC)。使用RT-PCR测定各组中lncRNAATP2B2-IT2和VEGF的表达水平。此后,细胞增殖，迁移,使用CCK-8，Transwell，和试管形成测定，分别。
结果：RT-PCR显示，HG组lncRNAATP2B2-IT2和VEGF的表达水平高于NG组（P<0.05）。lncRNAATP2B2-IT2沉默后,VEGF的表达显著降低(P<0.05)。随后的CCK-8,Transwell,和试管形成试验表明，与NG组相比，HG组的HRMEC表现出显著增加的增殖，迁移,新生血管形成(P<0.05)。然而,lncRNAATP2B2-IT2沉默后，增殖，迁移,与HG组相比，HGsi-lncRNAATP2B2-IT2组的HRMECs新生血管明显减少（P<0.05）。
结论：LncRNAATP2B2-IT2可能促进细胞增殖，高葡萄糖条件下HRMEC的迁移和新生血管形成。
