scholarly journals MicroRNA-126 modulates angiogenesis and tube formation through enhancing epidermal growth factor-like domain 7 expression and phosphorylating PI3K/AKT signaling pathway

2021 ◽  
Author(s):  
Qiang Li ◽  
Zhong-ming Wang ◽  
Ai-yue Wang ◽  
Qiong-Guan Xu ◽  
Zhou-feng Fu ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Signaling Pathway ◽  
Akt Signaling ◽  
Tube Formation ◽  
Luciferase Assay ◽  
Control Group ◽  
Akt Signaling Pathway ◽  
Invasive Ability ◽  
Epidermal Growth

IntroductionDysregulated angiogenesis is a critical characteristic for endothelial dysfunction disorders. This study aimed to determine functions of microRNA-126 in formation of tube and investigated the potential mechanisms.Material and methodsThe synthesized microRNA-126 control and microRNA-126 inhibitor plasmids were transfected into human umbilical-vein endothelial cells (HUVECs) using lipofectamine 2000 reagent. Cell counting kit-8 (CCK-8) was employed to measure proliferative capability of HUVECs. Transwell analysis was used to evaluate HUVECs invasive ability. Real time PCR (RT-PCR) was utilized to access epidermal growth factor-like domain 7 (EGFL7) and microRNA-126 mRNA transcription. Tube-forming capability in HUVECs was determined. Dual-luciferase assay and linear-regression analysis were conducted to measure interaction between EGFL7 and microRNA-126 molecule. Phosphoinositide-3-kinase/protein kinase-B (PI3K/AKT) signaling pathway associated molecules were evaluated using western blot assay.ResultsSilencing of microRNA-126 significantly enhanced proliferative capability and invasive ability of HUVECs compared to those of microRNA-126 control group (p<0.05). microRNA-126 silencing remarkably promoted tube formation and significantly up-regulated EGFL7 compared to those of microRNA-126 control group (p<0.05). microRNA-126 could interact with EGFL7 molecule. microRNA-126 was also negatively correlated with EGFL7 molecule in HUVECs (p<0.05). Silencing of microRNA-126 significantly enhanced p-PI3K/PI3K ratio compared to that of microRNA-126 control group (p<0.05). microRNA-126 silencing also remarkably increased p-AKT/AKT ratio compared to that of microRNA-126 control group (p<0.05).ConclusionsmicroRNA-126 modulated angiogenesis and tube formation through increasing EGFL7 expression and phosphorylating PI3K/AKT signaling pathway.

Knockdown of Nrf2 inhibits angiogenesis by downregulating VEGF expression through PI3K/Akt signaling pathway in cerebral microvascular endothelial cells under hypoxic conditions

2018 ◽  
Vol 96 (4) ◽  
pp. 475-482 ◽  
Author(s):  
Yujing Huang ◽  
Ying Mao ◽  
Huiying Li ◽  
Guangxun Shen ◽  
Guangxian Nan
Keyword(s):  
Vascular Endothelial Growth Factor ◽  
Ischemic Stroke ◽  
Growth Factor ◽  
Signaling Pathway ◽  
Akt Signaling ◽  
Tube Formation ◽  
Vascular Endothelial ◽  
Akt Signaling Pathway ◽  
Microvascular Endothelial ◽  
Endothelial Growth Factor

Ischemic stroke is a major cerebrovascular disease resulting from a transient or permanent local reduction of cerebral blood flow. Angiogenesis plays an important role in cerebral microvascular repair after ischemic stroke. This study aimed at investigating the effect of NF-E2-related factor 2 (Nrf2) on the angiogenesis of mouse cerebral microvascular endothelial bEnd.3 cells in a hypoxic environment. We found that Nrf2 expression was temporarily increased in hypoxia-induced bEnd.3 cells. Knockdown of Nrf2 inhibited the proliferation, migration, as well as tube formation in hypoxia-induced bEnd.3 cells. Meanwhile, vascular endothelial growth factor and PI3K/Akt signaling pathways were identified to be regulated by Nrf2 in hypoxia-induced bEnd.3 cells. It was found that silencing of Nrf2 downregulated the expression levels of NAD(P)H:quinine oxidoreductase-1, vascular endothelial growth factor, p-Akt, and heme oxygenase-1 in hypoxia-induced bEnd.3 cells. Data suggested that hypoxia induced the transient increase of Nrf2, which plays a key role in the angiogenesis of cerebral microangiogenesis, and that Nrf2 regulates the proliferation, migration, as well as tube formation likely through PI3K/Akt signaling pathway in hypoxia-induced bEnd.3 cells. Our study provides proof of concept for the modulation of Nrf2, so as to tilt the balance toward angiogenesis, representing a therapeutic strategy for hypoxia or ischemia disorders such as stroke.

scholarly journals Exosome-mediated miR-21 promotes angiogenesis within esophageal tumor microenvironment by activating PTEN/Akt signaling pathway in Vascular Endothelial Cells

2020 ◽  
Author(s):  
Tian Wang ◽  
Juan Liao ◽  
Zijie Yang ◽  
Weitao Shen ◽  
Zhi kui Gao ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Tumor Microenvironment ◽  
Signaling Pathway ◽  
Akt Signaling ◽  
Tube Formation ◽  
Luciferase Reporter ◽  
Control Group ◽  
Esophageal Tumor ◽  
Akt Signaling Pathway ◽  
Transwell Assay

Abstract Background Angiogenesis, a pivotal component in the tumor microenvironment (TME), boosts tumor growth and metastasis. Cancer-derived exosomes, which have been widely reported to play a crucial role in the establishment of TME can be effective angiogenic modulators. We aim to investigate the contribution of microRNA-21 (miR-21) for angiogenesis which was packaged in cancer-derived exosomes in esophageal squamous cell carcinoma (ESCC). Methods The co-cultivation model was constructed to mimic the tumor microenvironment based at a physical level to explore the effects of cancer-derived exosomes on angiogenesis of human umbilical vein endothelial cells (HUVECs). EdU assay, transwell assay and tube formation assay formation experiments were conducted for the evaluation of HUVECs proliferation, migration, and angiogenesis, respectively. In addition, Dual-luciferase reporter (DLR) assay was performed to validate the relationship between miR-21 and its target gene PTEN. Similarly, miR-21 inhibitors and LY294002 was applied to evaluate the regulation of miR-21 via pro-angiogenesis in recipient HUVECs by PTEN/Akt signaling pathway. Results After 24 h co-cultivation with EC9706 cells, miR-21 levels in recipient HUVECs was raised. The results from EdU assay, transwell assay and blood vessel formation experiment showed that exosomes which were secreted from EC9706 cells (EC9706-Exo) delivered miR-21 stimulated proliferation, migration and tube formation of HUVECs. DLR assay indicated that miR-21 could directly bind to the 3'-untranslated region (UTR) of PTEN genes, real-time PCR and western blot analysis for PTEN showed it was inhibited by EC9706-Exo shuttled miR-21. Meanwhile, phospho-Akt (p-Akt) (Ser473), one of the downstream genes of PTEN, was significantly increased in recipient HUVECs compared to the control group, while inhibiting miR-21 and PI3K/Akt pathway respectively both led to a sharp decrease in p-Akt levels, suggesting that exosomal miR-21 promote angiogenesis via activating PTEN/Akt signaling pathway. Conclusion Exosomal miR-21 acts as a driver of pro-angiogenesis by activating PTEN/Akt signaling pathway, it might serve as a blood-based biomarker for ESCC metastasis. Suppressing the expression or blocking the transmission of these exosome-derived miR-21 might be a novel antiangiogenic therapeutic strategy for ESCC.

scholarly journals CircRILPL1 promotes muscle proliferation and differentiation via binding miR-145 to activate IGF1R/PI3K/AKT pathway

2021 ◽  
Vol 12 (2) ◽  
Author(s):  
Xuemei Shen ◽  
Jia Tang ◽  
Rui Jiang ◽  
Xiaogang Wang ◽  
Zhaoxin Yang ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Muscle Development ◽  
Inhibitory Effect ◽  
Akt Signaling ◽  
Luciferase Assay ◽  
Circular Rnas ◽  
Akt Signaling Pathway ◽  
Sequencing Data ◽  
Proliferation And Differentiation ◽  
Myoblast Proliferation

AbstractMany novel non-coding RNAs, such as microRNAs (miRNAs) and circular RNAs (circRNAs), are involved in various physiological and pathological processes. The PI3K/AKT signaling pathway is important for its role in regulating skeletal muscle development. In this study, molecular and biochemical assays were used to confirm the role of miRNA-145 (miR-145) in myoblast proliferation and apoptosis. Based on sequencing data and bioinformatics analysis, we identified a new circRILPL1, which acts as a sponge for miR-145. The interactions between circRILPL1 and miR-145 were examined by bioinformatics, a luciferase assay, and RNA immunoprecipitation. Mechanistically, knockdown or exogenous expression of circRILPL1 in the primary myoblasts was performed to prove the functional significance of circRILPL1. We investigated the inhibitory effect of miR-145 on myoblast proliferation by targeting IGF1R to regulate the PI3K/AKT signaling pathway. A novel circRILPL1 was identified that could sponge miR-145 and is related to AKT activation. In addition, circRILPL1 was positively correlated with muscle proliferation and differentiation in vitro and could inhibit cell apoptosis. The newly identified circRILPL1 functions as a miR-145 sponge to regulate the IGF1R gene and rescue the inhibitory effect of miR-145 on the PI3K/AKT signaling pathway, thereby promoting myoblast growth.

Sign in / Sign up

Export Citation Format

Share Document