The role of microRNA‐93 regulating angiopoietin2 in the formation of malignant pleural effusion

Abstract

The biological roles of miRNAs in the development of malignant pleural effusion (MPE) are unclear. In this study, the miRNA microarray analysis was performed in two different prognosis groups of lung adenocarcinoma patients. Expression profiles of miRNAs in MPEs were identified. With the help of quantification PCR, we confirmed the expression differences of miRNAs and further analyzed their biological functions and relative target genes in vitro. The target gene of miR‐93 was estimated by online database, and also, the protein was tested. The target gene and the binding sites of specific miRNA were estimated by online database. The combining capacity of binding sites was verified by luciferase reporter gene assay, and the target gene protein was tested by western blot. We detected 107 miRNAs with expression differences (n = 10) and confirmed significant expression differences in miR‐93 and miR‐146a in two groups of patients (n = 84). By manipulating miR‐93 expression of human lymphatic endothelial cells (HLEC) and human umbilical vein endothelial cells (HUVEC), we discovered that high expression of miR‐93 inhibited migration, proliferation, and angiogenesis. And also, miR‐93 increased not only apoptosis, but also G1 phase cell block. By using luciferase reporter gene assay and western blot, we confirmed that angiopoietin2 (Ang2) was the target of miR‐93. The data showed that miR‐93 has an inhibiting effect on pleural effusion. By targeting Ang2, miR‐93 regulates angiogenesis and lymphangiogenesis and plays a role in pathogenetic mechanism of MPE. MiR‐93/Ang2 may shed light on potential new targets in cancer treatment.

Down‐regulation of exosomal miR‐106b‐5p derived from cholesteatoma perimatrix fibroblasts promotes angiogenesis in endothelial cells by overexpression of Angiopoietin 2

“…Luciferase assays confirmed that miR‐106b‐5p bound directly to the 3′‐UTR of Angpt2 mRNA and inhibited its translation. Moreover, consistent with results of previous studies, knock‐down of Angpt2 in endothelial cells significantly decreased their migration and tube formation (Jiang et al, ; Liang et al, ; Qian et al, ).…”

“…Expression of Angpt2 is normally low in quiescent mature vessels, and up‐regulation of Angpt2 expression in endothelial cells leads to initiation of angiogenesis (Ahmad et al, ). Recent studies have shown that inhibition of Angpt2 signaling decreases HUVEC migration, adhesion, and tube formation (Jiang et al, ; Liang et al, ; Qian et al, ). Here, we used miRNA target prediction algorithms to identify Angpt2 as a potential target of miR‐106b‐5p.…”

The regulatory role of microRNAs in angiogenesis‐related diseases

“…In neuroblastoma, miR‐93 represses IL‐8‐ and VEGF‐dependent angiogenesis by targeting IL‐8 and VEGF mRNAs 127. In addition, miR‐93 reduces angiogenesis in human lymphatic endothelial cells (HLECs) by reducing angiopoietin 2 (Ang 2) expression by directly binding to the 3′UTR of the Ang 2 gene 128. Taken together, miR‐93 has dual effects on angiogenesis in different tissues and cells via multiple molecular mechanisms.…”

“…MiR‐221 plays a major role in the regulation of the phenotypic change, differentiation and proliferation of VSMCs by inhibiting c‐kit and p27 (Kip1) mRNA expression, thereby modulating atherosclerosis remodelling 168. Moreover, miR‐221 and miR‐222 stimulates VSMC proliferation thereby accelerating neointima formation,128 which contributes to plaque progression. Reports have shown that miR‐21, miR‐146b, miR‐146a, miR‐210 and miR‐34a are up‐regulated in plaques 169.…”

MicroRNAs – novel biomarkers for malignant pleural effusions

“…The role of specific miRNA, particularly miRNA-93, has been elucidated in the regulation of endothelial cell migration, proliferation, angiogenesis, and cell cycle, playing an important part in the occurrence and development of malignant pleural effusions. Its high expression is associated with a good prognosis, due to its inhibition of pleural effusion by regulating angiogenesis and lymphangiogenesis [87]. Furthermore, the differing expression levels of miRNAs, such as miRNA-182 and miRNA-210 expression, are seen in malignant pleural effusions associated with lung adenocarcinoma compared to benign pleural effusion, providing a useful role in the minimally invasive screening for diagnostic evaluations of pleural effusions [88].…”