Cell Journal
·
2021
·
IF 2.479
Sadaf Mahboudi,Kazem Parivar,Zohreh Mazaheri,Shiva Irani
Objective Numerous evidence indicates that microRNAs (miRNAs) are critical regulators in the spermatogenesisprocess. The aim of this study was to investigate Mir-106b cluster regulates primordial germ cells (PGCs) differentiationfrom human mesenchymal stem cells (MSCs). Materials and Methods In this experimental study, samples containing male adipose (n: 9 samples- age: 25-40 years)were obtained from cosmetic surgeries performed for the liposuction in Imam Khomeini Hospital. The differentiationof MSCs into PGCs was accomplished by transfection of a lentivector expressing miR-106b . The transfection of miR-106b was also confirmed by the detection of a clear green fluorescent protein (GFP) signal in MSCs. MSCs weretreated with bone morphogenic factor 4 (BMP4) protein, as a putative inducer of PGCs differentiation, to induce thedifferentiation of MSCs into PGCs (positive control). After 4 days of transfection, the expression of miR-106b , STELLA,and FRAGILIS genes was evaluated by real-time polymerase chain reaction (PCR). Also, the levels of thymocytedifferentiation antigen 1 (Thy1) protein was assessed by the western blot analysis. The cell surface expression of CD90was also determined by immunocytochemistry method. The cytotoxicity of miR-106b was examined in MSCs after 24,48, and 72 hours using the MTT assay. Results MSCs treated with BMP4 or transfected by miR-106b were successfully differentiated into PGCs. The resultsof this study also showed that the expression of miR-106b was significantly increased after 48 hours from transfection.Also, we showed STELLA, FARGILIS , as well as the protein expression of Thy1, was significantly higher in MSCstransfected by lentivector expressing miR-106b in comparison with MSCs treated with BMP4 (P≤0.05). MTT assayshowed miR-106b was no toxic during 72 hours in 1 µg/ml dose, that this amount could elevated germ cells markersignificantly higher than other experimental groups (P≤0.05). Conclusion According to this findings, it appears that miR-106b plays an essential role in the differentiation of MSCsinto PGCs.
研究领域:
分子生物学
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生殖医学
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干细胞研究
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DOI
PubMed
高影响因子
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
·
2024
·
IF 6.2
Biyun Chen,Lujiao Wang,Luyao Li,Mei Zhou,Shuya Pan,Qin Wang,Yaxuan Hou,Xue Zhou
Arsenic is a widespread carcinogen and an important etiological factor for lung cancer. Dysregulated miRNAs have been implicated in arsenic carcinogenesis and the mechanisms of arsenic-induced dysregulated miRNAs have not been fully elucidated. N 6 -methyladenosine (m 6 A) modification is known to modulate pri-miRNA processing. However, whether m 6 A-mediated pri-miRNA processing is involved in arsenic carcinogenesis is poorly understood. Here, we found that m 6 A modification was significantly increased in arsenite-transformed human bronchial epithelial BEAS-2B cells (0.5 µM arsenite, 16 weeks). Meanwhile, METTL3 was significantly upregulated at week 12 and 16 during cell transformation. The proliferation, migration, invasion, and anchorage-independent growth of arsenite-transformed cells were inhibited by the reduction of m 6 A levels through METTL3 knockdown. Further experiments suggest that the oncogene miR-106b-5p is a potentially essential m 6 A target mediating arsenic-induced lung cancer. miR-106b-5p was observed to be upregulated after exposure to arsenite for 12 and 16 weeks, and the reduction of m 6 A levels caused by METTL3 knockdown inhibited miR-106b-5p maturation in arsenite-transformed cells. What’s more, miR-106b-5p overexpression successfully rescued METTL3 knockdown-induced inhibition of the neoplastic phenotypes of transformed cells. Additionally, Basonuclin 2 (BNC2) was uncovered as a potential target of miR-106b-5p and downregulated by METTL3 via enhancing miR-106b-5p maturation. Additionally, the METTL3 inhibitor STM2457 suppressed neoplastic phenotypes of arsenite-transformed BEAS-2B cells by blocking pri-miR-106b methylation. These results demonstrate that m 6 A modification promotes the neoplastic phenotypes of arsenite-transformed BEAS-2B cells through METTL3/miR-106b-5p/BNC2 pathway, providing a new prospective for understanding arsenic carcinogenesis.
研究领域:
癌症生物学
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分子生物学
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毒理学
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表观遗传学
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DOI
PubMed
高影响因子
Journal of Cardiovascular Translational Research
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2023
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IF 3.4
Wang Li,Wang Chuanhe,Sun Zhaoqing,Du Aolin,Shan Fei,Sun Zhijun
Myocardial ischemia/reperfusion (MI/R) injury induces myocardial damage and dysfunction. Increasing evidence has confirmed that circular RNAs (circRNAs) play crucial roles in regulating MI/R. Mmu-circ-0001380 has identified to be highly expressed in myocardium of MI/R mouse model. However, its biological function and molecular mechanism in MI/R injury are still unclear. Here, we demonstrated that knockdown of cric-0001380 attenuated myocardial injury of MI/R mice. In vitro , silence of circ-0001380 significantly enhanced viability, and inhibited apoptosis and oxidative stress in HL-1 cells under oxygen–glucose deprivation/reoxygenation (OGD/R). Mmu-miR-106b-5p interacted with circ-0001380, and suppressed the expression of pleckstrin homology domain and leucine rich repeat protein phosphatase 2 (Phlpp2). The miR-106b-5p/Phlpp2 axis mediated the effect of circ-0001380 on OGD/R-induced apoptosis through regulating the phosphorylation of p38, and further involved in regulating the viability and oxidative stress of HL-1 cells. In conclusion, circ-0001380 downregulation relieves MI/R injury via regulating the miR-106b-5p/Phlpp2 axis. Graphical Abstract The present study indicates that mmu-circ-0001380 exacerbates the myocardial ischemia/reperfusion injury through modulating the miR-106b-5p/Phlpp2 axis in vitro and in vivo .
研究领域:
心脏病学
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分子生物学
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细胞生物学
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PubMed
高影响因子
EXPERIMENTAL BRAIN RESEARCH
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2021
·Vol. 239
·期 11
·3315-3325
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IF 1.972
Yu Tao,Fu Hui,Sun Jing-Jing,Ding Dan-Rui,Wang Hua
To investigate the association of miR-106b-5p with neuroinflammation and microglial activation in a status epilepticus (SE) mouse model. We examined changes in the expression of microRNA-106b-5p (miRNA-106b-5p), repulsive guidance molecule A (RGMa), triggering receptor expressed on myeloid cells 2 (TREM2), and the microglia-related markers interleukin (IL)-1β, IL-4, IL-6, IL-10, inducible nitric oxide synthase (iNOS), and arginase-1 (Arg-1) in the mouse hippocampus of the lithium–pilocarpine-induced SE mouse model. Eighty-four female C57BL/6 mice were randomly divided into a normal control group ( n = 12), and six SE groups ( n = 12/group), which were monitored at 6 h and at 1, 3, 7, 14, and 21 days (d) post-SE induction. Unlike in the dentate gyrus, immunohistochemical staining revealed prominent neuronal swelling at 6 h, significant neuronal loss and apoptosis on day 3, and recovery by day 14 in the hippocampal cornu ammonis (CA)1 and CA3 pyramidal cells in SE mice. We noted elevated levels of miRNA-106b-5p and all microglia-related markers, which peaked at 3 days post-SE, except IL-4, which peaked at 7 days post-SE, indicating inflammation and microglial activation. RGMa and TREM2 levels decreased at 6 h post-SE. All markers but miRNA-106b-5p, RGMa, and TREM2 returned to baseline levels at 21 days post-SE. Dual luciferase reporter gene assay showed that microRNA-106b-5p can interact with RGMa. We observed that miR-106b-5p level increased while both RGMa and TREM2 levels decreased post-SE and showed associations with microglial activation and inflammation in the mouse hippocampus, suggesting their potential as SE therapeutic targets.
研究领域:
神经科学
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分子生物学
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神经炎症
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小胶质细胞
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癫痫
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2026
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IF 6.1
Ran Xu,Hao Chen,Chao Zhao,Xiqing Ding
The early, non-invasive diagnosis of Alzheimer's disease (AD) remains a significant clinical challenge. MicroRNAs carried by L1CAM-immunocaptured small extracellular vesicles (sEVs) in blood, particularly miR-106b-5p, have emerged as promising candidate biomarkers for AD. However, their detection requires methods that are sensitive, specific, stable, and suitable for point-of-care (POC) applications. Catalytic hairpin assembly (CHA) combined with lateral flow immunoassay (LFIA) offers a potential POC solution, but its practical application is limited by high background leakage caused by “DNA breathing” and poor probe stability. Herein, we propose an integrated POC platform that combines immunomagnetic enrichment of L1CAM-positive sEVs with a novel “Fold-Adsorb-Block-Immobilize-Lyophilize” (FABIL) process for CHA probe stabilization. The FABIL process utilizes magnetic graphene oxide (MGO) to adsorb and physically separate hairpin probes (H1 and H2), thereby suppressing nonspecific hybridization and generating lyophilized CHA microspheres with room-temperature stability. When integrated into a lateral flow test strip, the FABIL-NE-miR platform achieves a visual detection limit of 10 fM and a liquid-phase limit of detection of 1.14 fM, overcoming the traditional trade-off between low background and high sensitivity. Clinical validation using 100 serum samples, including 32 AD cases and 68 non-AD controls, demonstrated that FABIL-NE-miR outperformed traditional low-background CHA systems, with an area under the ROC curve (AUC) of 0.9563, a sensitivity of 90.63%, and a specificity of 94.12%. Overall, the FABIL-NE-miR platform enables rapid visual detection of low-abundance AD-associated miRNAs from L1CAM-immunocaptured sEV fractions. By addressing key challenges such as background leakage and probe instability, this study provides a practical tool for AD screening and supports the feasibility of the FABIL strategy as a universal platform for next-generation nucleic acid diagnostics.
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高影响因子
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