Resveratrol protects BV2 mouse microglial cells against LPS-induced inflammatory injury by altering the miR-146a-5p/TRAF6/NF-κB axis

Abstract

Objective: To investigate the role of miR-146a-5p in the effects of resveratrol (RSV) on inflammatory response in BV2 mouse microglial cells.

Materials and methods: BV2 cells were pretreated by RSV and stimulated with lipopolysaccharide (LPS). Cell Viability was checked using a MTT assay. Real-Time PCR was performed to detect the levels of pro-inflammatory cytokines (tumor necrosisfactor-α－TNF-α, interleukin-1β－IL-1β and interleukin-6 － IL-6) and miR-146a-5p expression. Western blot was used to analyze the protein expression of TNF receptor associated factor 6 (TRAF6) and phospho-nuclear factor kappa B (pNF-κB). Gain-of-function and loss-of-function analysis of miR-146a-5p was performed using transfection of miR-146a-5p mimic and miR-146a-5p inhibitor, respectively.

Results: Pretreatment with RSV significantly and dose dependently inhibited LPS-induced production of TNF-α, IL-1β and IL-6 in BV2 cells. MiR-146a-5p was significantly upregulated after LPS treatment, and further increased in RSV and LPS-co-treated cells. MiR-146a-5p overexpression via miR-146a-5p mimic transfection downregulated the mRNA level of TNF-α, IL-1β and IL-6, as well as abrogated the protein expression of TRAF6 and pNF-κB in BV2 cells exposed to LPS. More importantly, the reducion of TNF-α, IL-1β and IL-6 level by RSV were reversed by miR-146a-5p silence via miR-146a-5p inhibitor transfection. Furthermore, silencing miR-146a-5p attenuated the inhibitory effect of RSV on the TRAF6/NF-κB pathway which was activated after induction with LPS. Conclusions: RSV can suppress LPS-induced inflammatory injury via modulating the miR-146a-5p/TRAF6/NF-κB axis in BV2 mouse microglial cells.     

MiR-29b-3p affects growth and biological functions of human extravillous trophoblast cells by regulating bradykinin B2 receptor

IntroductionThis study investigated miR-29b-3p’s effects and mechanisms in preeclampsia development.Material and methodsIn this study, we analysed the pathology and expression of miR-29b-3p and B2R mRNA from normal and preeclampsia placenta tissues using hematoxylin and eosin staining and RT-qPCR assay. For cell experiments, we used transwell assay CCK-8, flow cytometry and wound healing assay to determine the effects and correlation of miR-29b-3p and B2R in HTR-8/SVneo cell proliferation, apoptosis, cell cycle, cell invasion and migration in a preeclampsia cell model. Moreover, the mechanisms were determined using Western blot or immunofluorescence in different groups.ResultsClinical analysis revealed that miR-29b-3p gene expression dramatically increased with increasing degree of preeclampsia (p < 0.001 or p < 0.05, respectively). The HTR-8/SVneo cell biological activities of the model group were significantly depressed (p < 0.001). However, with miR-29b-3p inhibitor or B2R transfection, the HTR-8/SVneo cell biological activities significantly recovered (p < 0.001). Western blot assay showed that B2R, VEGF-A, CCND-1, MMP-2 and MMP-9 levels were suppressed in the model group, compared with those in the NC groups (p < 0.001, respectively). With miR-29b-3p inhibitor or B2R transfection, the protein expression levels of B2R, VEGF-A, CCND-1, MMP-2 and MMP-9 dramatically increased (p < 0.001, respectively).ConclusionsThe down-regulation of miR-29b-3p could improve HTR-8/SVneo cell biological activities in a preeclampsia cell model by targeting B2R.     

miR-26b-5p suppresses proliferation and promotes apoptosis in multiple myeloma cells by targeting JAG1

Background

Though the levels of diagnosis and treatment of multiple myeloma (MM) have been largely improved recent years, the prognosis of these patients remain unacceptable. It is urgent for us to discover the exact mechanism and determine some new indicators for MM. MiRNAs play a critical role in the occurrence and progression of cancers, including MM. MiR-26b-5p has been reported to be closely related to cells proliferation in human pulmonary cancer, hepatocellular carcinoma and so on.

p53/miR-502-5p/TRAF2通路对骨关节炎软骨细胞损伤的影响

目的:探究微小RNA-502-5p(miR-502-5p)对白细胞介素-1β(IL-1β)诱导的骨关节炎(OA)软骨细胞损伤的影响。方法:Western blot和RT-q PCR检测骨关节炎患者软骨组织和IL-1β诱导的软骨细胞中p53和miR-502-5p的表达;分离培养软骨,并分组处理细胞,分别用MTT法检测细胞活力,流式细胞术检测细胞凋亡,Western blot检测炎性因子和细胞外基质(ECM)相关蛋白的表达,另外,萤光素酶报告实验检测miR-502-5p对p53及肿瘤坏死因子受体相关因子2(TRAF2)的调控作用。结果:与正常软骨组织相比,OA患者软骨组织中miR-502-5p的水平显著降低,p53和TRAF2水平则明显升高(P0.05)。IL-1β处理软骨细胞后,细胞活力下降、细胞凋亡率增加、IL-6、IL-8和TNF-α的蛋白表达水平显著上升;Ⅱ型胶原和蛋白聚糖的蛋白表达显著下调,基质金属蛋白酶(MMP)-3、MMP-9和MMP-13等的蛋白表达显著上调,miR-502-5p mimic转染反转了IL-1β对软骨细胞的这些作用。此外,p53与miR-502-5p之间存在负反馈调节,同时miR-502-5p能够靶向抑制TRAF2的水平。TRAF2沉默同样反转了IL-1β对软骨细胞增殖、凋亡、炎症反应以及ECM相关蛋白表达的影响。结论:调节p53/miR-502-5p/TRAF2通路能够减轻IL-1β诱导的骨关节软骨细胞损伤。     

MiR-494-3p Upregulation Exacerbates Cerebral Ischemia Injury by Targeting Bhlhe40

PurposeCerebral ischemia is related to insufficient blood supply and is characterized by abnormal reactive oxygen species (ROS) production and cell apoptosis. Previous studies have revealed a key role for basic helix-loop-helix family member e40 (Bhlhe40) in oxidative stress and cell apoptosis. This study aimed to investigate the roles of miR-494-3p in cerebral ischemia/reperfusion (I/R) injury.Materials and MethodsA mouse middle cerebral artery occlusion (MCAO/R) model was established to mimic cerebral ischemia in vivo. Brain infarct area was assessed using triphenyl tetrazolium chloride staining. Oxygen-glucose deprivation/reoxygenation (OGD/R) operation was adopted to mimic neuronal injury in vitro. Cell apoptosis was analyzed by flow cytometry. The relationship between miR-494-3p and Bhlhe40 was validated by luciferase reporter and RNA immunoprecipitation assays.ResultsBhlhe40 expression was downregulated both in MCAO/R animal models and OGD/R-induced SH-SY5Y cells. Bhlhe40 overexpression inhibited cell apoptosis and reduced ROS production in SH-SY5Y cells after OGD/R treatment. MiR-494-3p was verified to bind to Bhlhe40 and negatively regulate Bhlhe40 expression. Additionally, cell apoptosis and ROS production in OGD/R-treated SH-SY5Y cells were accelerated by miR-494-3p overexpression. Rescue experiments suggested that Bhlhe40 could reverse the effects of miR-494-3p overexpression on ROS production and cell apoptosis.ConclusionMiR-494-3p exacerbates brain injury and neuronal injury by regulating Bhlhe40 after I/R.     

MiR-103a-3p Contributes to the Progression of Colorectal Cancer by Regulating GREM2 Expression

PurposeOur research aimed to investigate the influence of miR-103a-3p on the growth and apoptosis of colorectal cancer (CRC) cells.Materials and MethodsBioinformatics was employed to analyze differentially expressed microRNAs and predict target genes. qRT-PCR was applied to detect the expression of miR-103a-3p in CRC and normal cells. HCT116 and Caco-2 were chosen, and miR-103a-3p mimics, miR-103a-3p inhibitor, as well as specific siRNAs targeting GREM2, were constructed. We subsequently evaluated alternations in cell proliferation, cell cycle and cell cycle regulators, apoptosis, and related proteins (Bcl-2 and Bax) by CCK-8 testing, Western blotting, luciferase reporter, colony formation, and Annexin V-FITC/PI. Possible binding sites for miR-103a-3p on the 3''UTR of GREM2 were checked with luciferase assay, and the impact of GREM2 on miR-103a-3p activity was also validated with above biological function testing. Additionally, the effect of miR-103a-3p knockdown in CRC cells and the molecular mechanism of miR-103a-3p targeting GREM2 were also studied.ResultsBioinformatics analysis revealed that miR-103a-3p expression increased remarkably in CRC, and targeted regulatory correlation existed between miR-103a-3p and GREM2. MiR-103a-3p inhibitor significantly impeded proliferative capacity and caused cell cycle arrest, as well as apoptosis, in HCT116 and Caco-2 cells. Consistent with this finding, overexpression of GREM2 showed similar effects to miR-103a-3p inhibition. Moreover, we demonstrated that miR-103a-3p connected target GREM2 and GREM2 knockdown reversed the effects of miR-103a-3p inhibitor on HCT116 and Caco-2 cell proliferation, cell cycle, and apoptosis. Further study showed that miR-103a-3p targeting GREM2 appeared to affect CRC progression via the transforming growth factor-β pathway.ConclusionMiR-103a-3p could augment CRC progression by targeting GREM2 and that miR-103a-3p/GREM2 could be potential novel targets for CRC therapy.     

CircCBFB is a mediator of hepatocellular carcinoma cell autophagy and proliferation through miR-424-5p/ATG14 axis

This study aims to investigate the role of circCBFB in hepatocellular carcinoma (HCC) cell proliferation and autophagy. qRT-PCR and Western blotting analyses quantified the expression levels of circCBFB, miR-424-5p, and ATG14 in HCC tissues and/or HCC cell lines. After transfection with pcDNA3.1-CircCBFB, sh-CircCBFB, miR-424-5p mimic, miR-424-5p inhibitor, pcDNA3.1-ATG14, sh-ATG14, sh-CircCBFB?+?miR-424-5p inhibitor, pcDNA3.1-CircCBFB?+?miR-424-5p mimic, sh-CircCBFB?+?pcDNA3.1-ATG14, or pcDNA3.1-CircCBFB?+?sh-ATG14, the proliferation, cell cycle, and apoptosis of Huh-7 and HCCLM3 cells were detected, respectively, through MTT assay and flow cytometry. Western blotting measured the expression levels of ATG14 and autophagy-related proteins (LC3-ΙΙ/LC3-Ι, Beclin1, and p62). The interactions among circCBFB, miR-424-5p, and ATG14 were identified through RNA fluorescence in situ hybridization and RNA immunoprecipitation. In HCC tissues, circCBFB and ATG14 were highly expressed, and miR-424-5p expression was downregulated. Transfection of pcDNA3.1-CircCBFB, miR-424-5p inhibitor, or pcDNA3.1-ATG14 into HCC cells facilitated HCC cell proliferation and autophagy, while suppressing cell apoptosis, evidenced by elevated cell viability, increased protein levels of autophagosome markers (LC3-ΙΙ/LC3-Ι and Beclin1), repressed apoptosis rate, and suppressed protein level of autophagy receptor p62. miR-424-5p was a target gene of circCBFB, and miR-424-5p negatively mediated ATG14. CircCBFB inhibits miR-424-5p and upregulates ATG14, thus promoting HCC cell proliferation and autophagy.

     

MiR-520b inhibits proliferation,migration and invasion in gallbladder carcinoma by targeting RAB22A

IntroductionPrevious studies have reported that miR-520b exhibited inhibitory effects on various human tumors, whereas the effects of miR-520b on gallbladder carcinoma (GBC) have remained unclear. To investigate the effects of miR-520b on GBC progression and reveal the underlying mechanisms, this study was performed.Material and methodsMiR-520b and RAB22A mRNA levels were analyzed by quantitative real-time PCR (qPCR). RAB22A protein level was analyzed via Western blot and immunohistochemical (IHC) analysis. The proliferation, colony formation ability, migration and invasion of NOZ cells were measured via MTT, colony formation, wound healing and transwell invasion assay respectively.ResultsMiR-520b expression level was lower in human GBC tissues than that in neighboring normal tissues. MiR-520b mimic repressed NOZ cell proliferation, colony formation ability, migration and invasion, whereas miR-520b inhibitor exhibited opposite effects. Dual luciferase reporter assay confirmed that miR-520b could bind to the 3′-untranslated regions of RAB22A mRNA. Moreover, RAB22A overexpression significantly abolished the anti-tumor effects of miR-520b in a NOZ cell model. Western blot, qPCR and IHC analysis proved that human GBC tissues showed a higher RAB22A expression level than neighboring normal tissues. Additionally, there was a negative association between miR-520b and RAB22A expression.ConclusionsMiR-520b had suppressive effects on GBC via targeting RAB22A in vitro.     

miR-26b-5p通过抑制REST表达调节胶质瘤细胞生长和侵袭

目的探究miR-26b-5p通过抑制抑制元素1-沉默转录因子(repressor element 1-silencing transcrip-tion factor,REST)表达调节胶质瘤细胞生长和侵袭情况.方法逆转录-聚合酶链反应检测miR-26b-5p和REST表达水平;荧光素酶报告实验检测miR-26b-5p和REST的靶向关系;Hoechst染色检测细胞凋亡;Colony形成法检测细胞增殖;Transwell检测细胞侵袭;蛋白免疫印迹检测细胞Ki67、MMP-9、cleaved caspase-3蛋白表达水平.结果miR-26b-5p和REST存在直接靶向作用关系,与Control组比较,mimic组细胞增殖数、侵袭数和Ki67、MMP-9表达明显降低,凋亡率与cleaved caspase-3表达明显升高,inhibitor组细胞增殖数、侵袭数和Ki67、MMP-9表达明显升高,凋亡率与cleaved caspase-3表达明显降低.结论miR-26b-5p可以通过抑制对REST的表达调节胶质瘤细胞生长、侵袭、凋亡能力.     

MicroRNA-574-5p directly targets FOXN3 to mediate thyroid cancer progression via Wnt/β-catenin signaling pathway

ObjectiveThyroid cancer is the most common endocrine tumor. A large number of thyroid cancer-related miRNAs have been studied and identified. However, the detailed roles of miR-574-5p in thyroid cancer remain poorly understood. This work mainly aimed to investigate the role of miR-574-5p/FOXN3 axis and its mechanism in the thyroid cancer progression.MethodsMiR-574-5p, FOXN3, Wnt/β-catenin pathway, and apoptosis-related markers were measured by quantitative real-time PCR (qRT-PCR) and western blotting analysis, respectively. Luciferase reporter assay was employed to validate the direct targeting of FOXN3 by miR-574-5p. MTT, flow cytometry, wound healing and transwell experiments were applied to analyze the functions of FOXN3 and miR-574-5p in thyroid cancer cells.ResultsKnockdown of miR-574-5p up-regulated FOXN3 expression and miR-574-5p directly targeted FOXN3 in thyroid cancer cells. Biological function experiments showed that knockdown of miR-574-5p inhibited proliferation, migration, invasion and promoted apoptosis of thyroid cancer cells. The activation of Wnt/β-catenin pathway was suppressed by MiR-574-5p silencing. FOXN3 silencing reversed the effects of miR-574-5p inhibitor on FOXN3 level and Wnt/β-catenin singling pathway, also reversed the effects on cell migration, proliferation, invasion and apoptosis.ConclusionThe miR-574-5p/FOXN3 axis is a novel molecular mechanism that promotes thyroid cancer progression, suggesting their potential for clinical therapy of thyroid cancer.     

Circulating miR-29c-3p is downregulated in patients with acromegaly

Background/aimmiRNAs control various biological functions, such as cell proliferation, differentiation, signaling pathways, apoptosis and metabolism. Recently, it has been shown that there is a relationship between changes in miRNA expression and the development of acromegaly. Studies are needed to identify new disease-specific miRNAs. The aim of the current study is to evaluate plasma miR-29c-3p, miR-31-5p and miR-18a-5p steady-state levels in acromegaly. Another aim is to investigate whether there is a difference in the levels of these miRNAs in patients with inadequate control and controlled acromegaly with somatostatin analog (SSA) therapy. These miRNAs targeting the IGF-1 gene were determined by in silico estimation.Materials and methodsThe study included 30 healthy controls (HC) and 20 patients with acromegaly. Anterior pituitary functions and disease activities of patients with acromegaly were evaluated at the time of study. The miR-29c-3p, miR-31-5p and miR-18a-5p levels were measured using quantitative real-time PCR (RT-qPCR).ResultsThe expression level of miR-29c-3p was significantly lower in patients with acromegaly compared to the HC group (p < 0.001). This downregulation was more pronounced in patients with inadequately controlled acromegaly than in patients with acromegaly controlled with somatostatin analogues (SSA) therapy (p = 0.016). Univariate logistic regression analysis results showed that down regulation of miR-29c-3p expression increases the risk of developing acromegaly [OR (95% Cl) = 1.605 (1.142-2.257), p = 0.006]. There was no significant difference between the groups in terms of miR-31-5p and miR-18a-5p expression levels (p = 0.375 and p = 0.649, respectively). ConclusionPlasma miR-29c-3p expression level is downregulated in patients with acromegaly, and this is more pronounced in patients with inadequate control.     

Ginsenoside Rh2 inhibits thyroid cancer cell migration and proliferation via activation of miR-524-5p

IntroductionThyroid cancer is an important disease that threatens the health of humans. Ginsenoside Rh2 is known as an anticancer molecule; however, its function in thyroid cancer cells has not been reported. In the present study, we identified that Rh2 treatment of the thyroid cancer cell line K1 inhibited cell migration and proliferation.Material and methodsWe determined the Rh2 function in thyroid cancer cell lines. By RT-PCR, expression of miR-524-5p and related genes were determined. The cell phenotype including cell migration and proliferation were detected after serials treatment. The relevant protein level were checked by Western blot.ResultsInterestingly, we observed that miR-524-5p, a type of miRNA, had lower expression in the thyroid cancer cell lines TPC-1, K1, and NPA than in the normal thyroid cell line Nthyri3-1. Additionally, Rh2 treatment induced miR-524-5p expression. Further examination using overexpression of miR-524-5p identified that the miR-524-5p mimic inhibited cell migration and proliferation of the K1 line. Similar to Rh2-treated cells, the miR-524-5p mimic-expressing cells had increased E-cadherin and reduced vimentin levels compared to the control cells. Next, we examined the relationship between Rh2 and miR-524-5p with respect to thyroid cell migration and proliferation. Treatment with Rh2 and miR-524-5p inhibitor suppressed Rh2 action on K1 thyroid cell migration and proliferation, and the rates were similar to those in control cells, suggesting that Rh2 might induce miR-524-5p expression to inhibit thyroid cancer cell migration and proliferation.ConclusionsOur analyses identified Rh2 and miR-524-5p action on thyroid cancer cell migration and proliferation as well as the linkage between Rh2 and miR-524-5p in thyroid cancer cell development.     

miR-892a靶向调控SOX5对前列腺癌细胞增殖和迁移能力的影响

目的:探讨miR-892a对前列腺癌细胞的增殖和迁移能力的影响。方法:从癌症基因组图谱(TCGA)数据库下载499例前列腺肿瘤及52例癌旁组织的微小RNA(miRNA)的样本资料,再采用R语言的limma包及gplots包进行差异表达分析,绘制火山图和热图,筛选目的基因。采用荧光定量反转录聚合酶链式反应(RT-PCR)...     

lncRNA MALAT1靶向调控miR-146b-5p影响膀胱癌细胞侵袭和迁移

目的:探讨长链非编码RNA(lncRNA)肺腺癌转移相关转录因子1(MALAT1)靶向微小RNA-146b-5p(miR-146b-5p)影响膀胱癌细胞侵袭和迁移的机制。方法:在膀胱癌BIU-87细胞中转染MALAT1 siRNA,以real-time PCR方法测定转染效果,Transwell法测定侵袭及迁移能力,Western blot法检测细胞中上皮-间充质转化(EMT)相关蛋白波形蛋白(vimentin)、上皮型钙黏蛋白(E-cadherin)和迁移侵袭相关蛋白基质金属蛋白酶-2(MMP-2)蛋白表达的变化。生物信息学软件预测MALAT1与miR-146b-5p有靶向互补位点,利用双萤光素酶报告系统鉴定靶向关系。用real-time PCR方法检测下调MALAT1后BIU-87细胞中miR-146b-5p表达的变化。将MALAT1 siRNA和miR-146b-5p inhibitor共转染至BIU-87细胞中,用上述方法分析细胞侵袭、迁移及vimentin、E-cadherin和MMP-2蛋白表达的变化。结果:转染MALAT1 siRNA可明显下调BIU-87细胞中MALAT...     

miR-146b-5p promotes VSMC proliferation and migration

Vascular smooth muscle cells (VSMCs) play pivotal roles in the development of vascular diseases. While microRNAs are important in vascular pathologies, a few is known about their functional roles in VSMC phenotypes. We profiled microRNA expression in PDGF-BB treated VSMCs and found microRNA-146b-5p (miR-146b-5p) was upregulated. Inhibition of miR-146b-5p blocked in response to PDGF while reducing VSMC proliferation and migration. These studies implicate miR-146b-5p as necessary for PDGF-induced VSMC phenotype transition. Downstream miR-146b-5p targets modulating VSMC phenotypes will be further identified. Our study will help to understand the role of VSMCs in the pathology of vascular diseases.     

miR-766-5p 过表达介导 MMP-7 对胶质瘤细胞增殖、 侵袭与 凋亡的影响及机制

目的 探讨 miR-766-5p 对胶质瘤细胞增殖、 侵袭与凋亡的影响。 方法 Targetscan 预测 miR-766- 5p 与基质金属蛋白酶-7 (MMP-7) 靶向关系并进行验证, 构建 miR-766-5p 过表达、 MMP-7 过表达和 miR766-5p + MMP-7 过表达胶质瘤 U87 细胞系, BrdU 染色、 流式细胞法、 Transwell 实验、 Western 印迹分别检 测细胞增殖、 凋亡、 侵袭及相关蛋白表达。 结果 miR-766-5p 可靶向抑制 MMP-7 表达, miR-766-5p 过表达 可抑制 Ki67、 增殖细胞核抗原 (PCNA)、 Wnt1、 β-链蛋白 (β-catenin) 蛋白表达以抑制 U87 细胞增殖和侵 袭, 可促进 Bcl 相关 X 蛋白 (Bax) / B 细胞淋巴瘤/ 白血病-2 (Bcl-2) 蛋白表达以促进细胞凋亡, MMP-7 过表达可以缓解 miR-766-5p 过表达所致 U87 细胞增殖、 侵袭抑制作用和凋亡促进作用。 结论 miR-766-5p 可靶向抑制 MMP-7 表达抑制 U87 细胞胞增殖、 侵袭和促细胞凋亡。     

MiR-322-5p Alleviates Cell Injury and Impairment of Cognitive Function in Vascular Dementia by Targeting TSPAN5

PurposeAs the population ages, the incidence of clinical dementia has been rising around the world. It has been reported that microRNAs act as key diagnostic biomarkers and targets for various neurological conditions, including dementia. MiR-322-5p has been revealed to play an important role in multiple diseases. In this study, we aimed to investigate the role and regulatory mechanism of miR-322-5p in vascular dementia.Materials and MethodsIn this study, neonatal rat neurons (NRNs) were subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) to induce cell injury. The animals were subjected to permanent bilateral occlusion of the carotid arteries (2-vessel occlusion, 2VO) to induce the model of chronic brain hypoperfusion.ResultsMiR-322-5p expression was significantly downregulated in the neurons exposed to OGD/R and the hippocampi of 2VO rats. Overexpression of miR-322-5p ameliorated cell apoptosis and the inflammatory response in vitro. In a mechanistic study, miR-322-5p was confirmed to directly target and negatively regulate tetraspanin 5 (TSPAN5) in cultured NRNs. Moreover, overexpression of TSPAN5 could counteract the effects of miR-322-5p overexpression on cell apoptosis and the inflammatory response in OGD/R-treated neurons. More importantly, miR-322-5p improved cognitive ability and inhibited inflammatory production in 2VO rats.ConclusionOverall, the results suggest that miR-322-5p alleviates vascular dementia development by targeting TSPAN5. This discovery may provide a potential therapeutic target for dementia.     

LncRNA OGFRP1 promotes angiogenesis and epithelial-mesenchymal transition in colorectal cancer cells through miR-423-5p/CTCF axis

ObjectiveTo investigate the mechanism of lncRNA OGFRP1 affecting angiogenesis and epithelial-mesenchymal transition (EMT) in colorectal cancer (CRC) and provide a new target for the treatment of CRC.MethodsThe expressions of OGFRP1, miR-423-5p, and CTCF were measured in CRC cell lines (HT29, LoVo, HCT116, SW620, and SW480) and normal colonic epithelial cells NCM460. Gain and loss of function experiments were performed on HCT116 and SW620 cells, after which the proliferation, apoptosis, EMT, invasion, and migration of the cells were measured using CCK-8 and colony formation assays, flow cytometry, Western blotting, Transwell, and scratch assay. The transfected cells were incubated with human umbilical vein endothelial cells (HUVECs) to assess angiogenesis using tube formation assay. ELISA was performed to detect VEGF in the conditioned medium of HCT116 and SW620 cells. The interactions among OGFRP1, CTCF and miR-423-5p were validated by dual-luciferase reporter assay.ResultsCRC cell lines had increased expression levels of OGFRP1 and CTCF and a suppressed expression level of miR-423-5p when compared with NCM460 cells. Suppression on OGFRP1 or CTCF and overexpression of miR-423-5p led to inhibited proliferation, EMT, invasion and migration and increased apoptosis of HCT116 and SW620 cells. HUVECs incubated with cells transfected with si-OGFRP1, si-CTCF or miR-423-5p mimic had suppressed angiogenesis ability. The effect of OGFRP1 suppression in CRC cells could be counteracted by miR-423-5p inhibition. Both CTCF and OGFRP1 could bind to miR-423-5p.ConclusionOGFRP1 promotes proliferation, EMT, and angiogenesis in CRC through miR-423-5p/CTCF axis.