MiR-674-5p Suppresses the Proliferation and Migration of Glioma Cells by Targeting Cul4b

Neurochemical Research - Glioma multiforme (GBM) is the most common malignant primary brain tumors. Despite the considerable advances in GBM treatment, it is still one of the most lethal forms of...     

MiR-508-5p Inhibits the Progression of Glioma by Targeting Glycoprotein Non-metastatic Melanoma B

Glioma is a severe and highly lethal brain cancer, a malignancy largely stemming from growing in a relatively restrained area of the brain. Hence, the understanding of the molecular regulation of the growth of glioma is critical for improving its treatment. MicroRNA has become a hotspot in research on diseases, especially in the initiation and progression of different types of cancer. However, the molecular function and mechanisms of miR-508-5p in gliomagenesis are still unclear. The aim of this study was to investigate miR-508-5p expression in glioma and determine its effects on proliferation. miR-508-5p expression levels, both in glioma cell lines and in tissue, were significantly lower than in a normal human astrocyte cell line or adjacent tissues. Cell growth was analyzed using a MTT assay and over-expression of miR-508-5p was found to decrease glioma cell growth. Moreover, a bioinformatic analysis was performed, showing that glycoprotein non-metastatic melanoma B (GPNMB) was a direct target for miR-508-5p in glioma cells. Furthermore, in vivo treatment with miR-508-5p reduced GPNMB protein levels in the tumor. Additionally, overexpression of GPNMB without 3′-UTR partially reversed the cell growth arrest induced by miR-508-5p over-expression in glioma cells. In conclusion, these results indicate that increased expression of miR-508-5p might be related to glioma progression, indicating a potential role of miR-508-5p for clinical therapy.     

MiR-125a-3p Regulates Glioma Apoptosis and Invasion by Regulating Nrg1

The current study was designed to examine the functional role and mechanism of miR-125a-3p in glioma development. Quantitative RT-PCR was used to evaluate miR-125a-3p expression in 60 glioma cases of different malignant grades. Then, the clinic pathologic significance of miR-125a-3p expression was determined in combination with the prognosis of the patients. In addition, the effects and mechanisms of miR-125a-3p on the proliferation, apoptosis and invasion of glioma cells were further investigated. The results showed that the expression of miR-125a-3p was decreased significantly in most malignant glioma samples relative to normal brain tissues and glioma tissues of low-malignant degree. Further kaplan-meier survival analysis showed that the lower expression of miR-125a-3p was associated with a poor prognosis of GBM patients. Functional analysis showed that the reintroduction of miR-125a-3p into glioblastoma cell lines induces markedly the apoptosis and suppresses the proliferation and migration of glioblastoma cells in vitro and in vivo. Luciferase assay and Western blot analysis revealed that Nrg1 is a direct target of miR-125a-3p. Furthermore, an increased expression of Nrg1 could reverse the effects of overexpression of miR-125a-3p on the proliferation, apoptosis and migration of glioblastoma cells. These findings suggest that miR-125a-3p performed an important role in glioma development mediated by directly regulating the expression of Nrg1. This study also provides a potential target for diagnosis and treatment of malignant glioma.     

MiR-378a-5p直接靶ZEB1向上调鼻咽癌CNE-1细胞E-cadherin表达*

目的： Mir-378a-5p是一种被发现多年的微小RNA,其在包括肺癌、结肠癌和乳腺癌等多种肿瘤中都被认为具有抑制肿瘤生 长的作用。Mir-378a-5p与细胞增殖的关系在多篇文章中已经有较为详细的阐述,然而,目前没有报道提及miR-378a-5p是否通过 作用于细胞迁移和细胞粘附途径从而达到抑制肿瘤生长的作用。方法与结果：在本研究中,我们通过wound healing 和trans-well 的方法发现在鼻咽癌细胞CNE-1 中过表达miR-378a-5p显著的抑制了细胞迁移以及细胞浸润的过程。通过免疫印迹方法,我们 揭示了细胞粘附因子E-cadherin在过表达miR-378a-5p后显著上调。通过生物信息学的方法,我们预测了miR-378a-5p的可能作 用靶点,并通过双荧光报告载体的方法证实了ZEB1是miR-378a-5p的直接靶点。结论：我们的研究提示了miR-378a-5p造成的E-cadherin 的上调是通过直接抑制E-cadherin的负调控因子ZEB1造成的。E-cadherin的上调不但影响了细胞的迁移和粘附,而且 通过间接阻断Wnt通路抑制了下游控制细胞增殖的基因表达。本研究为理解miR-378a-5p的肿瘤抑制作用提供了一个新的作用 机理。     

Mir-378a-5p抑制鼻咽癌细胞系CNE-1 细胞增殖以及肿瘤生长的初步研究*

目的： MiR-378a-5p是一种被认为在多种肿瘤发生过程中具有抑制肿瘤生长的微小RNA。然而miR-378a-5p在鼻咽癌中的 作用尚未见报道。因此,本文旨在通过临床样本的miRNA 表达谱分析以及细胞学实验从而揭示miR-378a-5p在鼻咽癌肿瘤发生过程中的作用。方法与结果：我们通过生物信息学的方法获取了鼻咽癌临床样本中miR-378a-5p的表达信息并通过与正常组织的 对比发现miR-378a-5p在鼻咽癌肿瘤组织中表达水平显著降低（P<0.01）。其次,我们发现高表达miR-378a-5p的鼻咽癌CNE-1 细 胞增殖速度显著较对照组降低（约40%~50%）。克隆形成实验证实了瞬时转染miR-378a-5p的鼻咽癌CNE-1 细胞的克隆形成数 量显著减弱。我们通过将稳定表达miR-378a-5p的CNE-1 细胞注射到裸鼠体内形成移植瘤并记录肿瘤生长曲线,结果显示 miR-378a-5p高表达组的裸鼠移植瘤体积明显较对照组小约50%,肿瘤重量显著降低(对照组0.33 g,处理组0.15 g)。结论：本研究通过对临床样本的分析以及在细胞和动物水平的实验验证揭示了miR-378a-5p具有抑制鼻咽癌肿瘤细胞增殖和肿瘤生长的作用。     

MiR-30a-3p 对人滋养肿瘤细胞系JEG-3 细胞侵袭功能的影响

目的：MiRNAs 对于胎盘的形成和正常妊娠的维持起着至关重要的作用,它在胎盘中的表达失衡的可能导致了妊娠相关疾 病的发生,我们前期研究发现miR-30a-3p 在子痫前期患者胎盘上特异性高表达,推测miR-30a-3p 可能参与了子痫前期的发生发 展过程,本课题通过观察miR-30a-3p 对人滋养肿瘤细胞系JEG-3 细胞侵袭能力的影响,深入探讨miR-30a-3p 在子痫前期发病过 程中的作用。方法：应用瞬时转染技术在人滋养肿瘤细胞系JEG-3 细胞中分别转染miR-30a-3p mimics、mimics NC为miR-30a-3p 过表达组和阴性对照组,空白转染组为空白对照组,利用荧光实时定量PCR 技术检测各组细胞中miR-30a-3p的表达,Transwell 实验检测各组细胞侵袭能力的差别。结果：荧光实时定量PCR结果显示miR-30a-3p 过表达组与阴性对照组、空白对照组相比 miR-30a-3p 的表达量明显升高,差异具有统计学意义（P<0.05）;Transwell 实验结果显示miR-30a-3p过表达组细胞的侵袭能力与 阴性对照组、空白对照组相比均有降低,差异具有统计学意义(P<0.05)。阴性对照组与空白对照组的侵袭能力差异无统计学意义 (P>0.05)。结论：miR-30a-3p 可以显著下调JEG-3 细胞的侵袭力, miR-30a-3p 有可能通过降低滋养细胞的浸润能力,导致滋养细胞 对子宫肌层和螺旋动脉的浸润不足,造成“胎盘浅着床”,从而在子痫前期的发病过程中发挥了重要的作用,miR-30a-3p 有望成为 诊治子痫前期疾病的靶点。     

MiR-30a-3p对人滋养肿瘤细胞系JEG-3细胞侵袭功能的影响

目的：MiRNAs对于胎盘的形成和正常妊娠的维持起着至关重要的作用,它在胎盘中的表达失衡的可能导致了妊娠相关疾病的发生,我们前期研究发现miR-30a-3p在子痫前期患者胎盘上特异性高表达,推测miR-30a-3p可能参与了子痫前期的发生发展过程,本课题通过观察miR-30a-3p对人滋养肿瘤细胞系JEG-3细胞侵袭能力的影响,深入探讨miR-30a-3p在子痫前期发病过程中的作用。方法：应用瞬时转染技术在人滋养肿瘤细胞系JEG-3细胞中分别转染miR-30a-3p mimics、mimics NC为miR-30a-3p过表达组和阴性对照组,空白转染组为空白对照组,利用荧光实时定量PCR技术检测各组细胞中miR-30a-3p的表达,Transwell实验检测各组细胞侵袭能力的差别。结果：荧光实时定量PCR结果显示miR-30a-3p过表达组与阴性对照组、空白对照组相比miR-30a-3p的表达量明显升高,差异具有统计学意义（P〈0.05）;Transwell实验结果显示miR-30a-3p过表达组细胞的侵袭能力与阴性对照组、空白对照组相比均有降低,差异具有统计学意义（P〈0.05）。阴性对照组与空白对照组的侵袭能力差异无统计学意义（P〉0.05）。结论：miR-30a-3p可以显著下调JEG-3细胞的侵袭力,miR-30a-3p有可能通过降低滋养细胞的浸润能力,导致滋养细胞对子宫肌层和螺旋动脉的浸润不足,造成＂胎盘浅着床＂,从而在子痫前期的发病过程中发挥了重要的作用,miR-30a-3p有望成为诊治子痫前期疾病的靶点。     

Rosiglitazone Suppresses Glioma Cell Growth and Cell Cycle by Blocking the Transforming Growth Factor-Beta Mediated Pathway

Glioma is one of the most malignant tumors in the central nervous system. As a peroxisome proliferator-activated receptor γ (PPAR-γ) activator, the thiazolidinediones (TZDs) induce growth arrest and cell death in a broad spectrum of tumor cells. In this study, we investigated the role of rosiglitazone in glioma cells. We found that rosiglitazone, a member of TZDs, suppresses growth of human glioma cell lines U87 and U251. Rosiglitazone also induces cell cycle arrest and apoptosis, which may be the mechanism of its anti-proliferation effect. Next, we found that rosiglitazone suppresses the expression of TGF-beta and its receptor TGF-betaR2, and suppresses phosphorylation of Smad3. Rosiglitazone also inhibits formation of the Smad3/Smad4 complex. Furthermore, Rosiglitazone affects the expression of Smad3/Smad4 associated regulators of gene expression, including p21 and c-Myc. These results suggest that rosiglitazone suppresses growth and cell cycle of human glioma cells by blocking the TGF-beta mediated pathway.     

MiR-199a Regulates Cell Proliferation and Survival by Targeting FZD7

A growing amount of evidence indicates that miRNAs are important regulators of multiple cellular processes and, when expressed aberrantly in different types of cancer such as hepatocellular carcinoma (HCC), play significant roles in tumorigenesis and progression. Aberrant expression of miR-199a-5p (also called miR-199a) was found to contribute to carcinogenesis in different types of cancer, including HCC. However, the precise molecular mechanism is not yet fully understood. The present study showed that miR-199a is frequently down-regulated in HCC tissues and cells. Importantly, lower expression of miR-199a was significantly correlated with the malignant potential and poor prognosis of HCC, and restoration of miR-199a in HCC cells led to inhibition of the cell proliferation and cell cycle in vitro and in vivo. Furthermore, Frizzled type 7 receptor (FZD7), the most important Wnt receptor involved in cancer development and progression, was identified as a functional target of miR-199a. In addition, these findings were further strengthened by results showing that expression of FZD7 was inversely correlated with miR-199a in both HCC tissues and cells and that over-expression of miR-199a could significantly down-regulate the expression of genes downstream of FZD7, including β-catenin, Jun, Cyclin D1 and Myc. In conclusion, these findings not only help us to better elucidate the molecular mechanisms of hepatocarcinogenesis from a fresh perspective but also provide a new theoretical basis to further investigate miR-199a as a potential biomarker and a promising approach for HCC treatment.     

MiR-467a-5p aggravates myocardial infarction by modulating ZEB1 expression in mice

Journal of Molecular Histology - Myocardial infarction (MI) is a great threat to patients all over the word. MicroRNAs (miRNAs) are a group of non-coding RNAs and can regulate initiation and...     

MicroRNA MiR-214 Regulates Ovarian Cancer Cell Stemness by Targeting p53/Nanog

Previous studies have shown aberrant expression of miR-214 in human malignancy. Elevated miR-214 is associated with chemoresistance and metastasis. In this study, we identified miR-214 regulation of ovarian cancer stem cell (OCSC) properties by targeting p53/Nanog axis. Enforcing expression of miR-214 increases, whereas knockdown of miR-214 decreases, OCSC population and self-renewal as well as the Nanog level preferentially in wild-type p53 cell lines. Furthermore, we found that p53 is directly repressed by miR-214 and that miR-214 regulates Nanog through p53. Expression of p53 abrogated miR-214-induced OCSC properties. These data suggest the critical role of miR-214 in OCSC via regulation of the p53-Nanog axis and miR-214 as a therapeutic target for ovarian cancer.     

MiR-519d-3p Suppresses Invasion and Migration of Trophoblast Cells via Targeting MMP-2

Our study was approved by the Medical Ethics Committee of Tang Du Hospital, Fourth Military Medical University and complied strictly with national ethical guidelines. Preeclampsia (PE) is a specific clinical disorder characterized by gestational hypertension and proteinuria and is a leading cause of maternal and perinatal mortality worldwide. The miR-519d-3p is upregulated in the maternal plasma of patients with PE which indicates a possible association between this microRNA and the pathogenesis of PE. No studies to date have addressed the effect of miR-519d-3p on the invasion and migration of trophoblast cells. In our study, we found that miR-519d-3p expression was elevated in placental samples from patients with PE. In vitro, overexpression of miR-519d-3p significantly inhibited trophoblast cell migration and invasion, whereas transfection of a miR-519d-3p inhibitor enhanced trophoblast cell migration and invasion. Luciferase assays confirmed that matrix metalloproteinase-2 (MMP-2) is a direct target of miR-519d-3p. Quantitative real-time PCR and western blot assays showed that overexpression of miR-519d-3p downregulated MMP-2 mRNA and protein expression. Knockdown of MMP-2 using a siRNA attenuated the increased trophoblast migration and invasion promoted by the miR-519d-3p inhibitor. In placentas from patients with PE or normal pregnancies, a negative correlation between the expression of MMP-2 and miR-519d-3p was observed using the Pearson correlation and linear regression analysis. Our present findings suggest that upregulation of miR-519d-3p may contribute to the development of PE by inhibiting trophoblast cell migration and invasion via targeting MMP-2; miR-519d-3p may represent a potential predictive and therapeutic target for PE.     

MiR-101a-3p Attenuated Pilocarpine-Induced Epilepsy by Downregulating c-FOS

This study aimed to explore the effects and function of microRNA-101a-3p (miR-101a-3p) in epilepsy. Rat model of pilocarpine-induced epilepsy was established and the seizure frequency was recorded. Expression of miR-101a-3p and c-Fos in hippocampus tissues of Rat models were detected by qRT-PCR and western blot. Besides, we established a hippocampal neuronal culture model of acquired epilepsy using Mg²⁺ free medium to evaluate the effects of miR-101a-3p and c-Fos in vitro. Cells were transfected with miR-101a-3p mimic, si-c-FOS, miR-101a-3p?+?c-FOS and its corresponding controls. MTT assay was used to detect cell viability upon transfection. Flow cytometry was performed to determine the apoptosis rate. Western blot was performed to measure the protein expression of apoptosis-related proteins (Bcl-2, Bax, and cleaved caspase 3), autophagy-related proteins (LC3 and Beclin1) and c-FOS. The targeting relationship between miR-101a-3p and c-FOS was predicted and verified by TargetScan software and dual-luciferase reporter assay. The role of miR-101a-3p was validated using epilepsy rat models in vivo. Another Rat models of pilocarpine-induced epilepsy with miR-NC or miR-101a-3p injection were established to evaluate the effect of miR-101a-3p overexpression on epilepsy in vivo. MiR-101a-3p was downregulated while c-FOS was increased in hippocampus tissues of Rat model of pilocarpine-induced epilepsy. Overexpression of miR-101a-3p or c-FOS depletion promoted cell viability, inhibited cell apoptosis and autophagy. C-FOS was a target of miR-101a-3p and miR-101a-3p negatively regulated c-FOS expression to function in epilepsy. Overexpression of miR-101a-3p attenuated pilocarpine-induced epilepsy in Rats in vivo. This study indicated that miR-101a-3p could attenuate pilocarpine-induced epilepsy by repressing c-Fos expression.