High-Resolution Fluorescence In Situ Hybridization to Detect mRNAs in Neuronal Compartments In Vitro and In Vivo

Author(s):  
Sharon A. Swanger ◽  
Gary J. Bassell ◽  
Christina Gross
2021 ◽  
Author(s):  
Penghui Xu ◽  
Xing Zhang ◽  
Jiacheng Cao ◽  
Jing Yang ◽  
Zetian Chen ◽  
...  

Abstract Background: Gastric cancer (GC) ranks third in motality among all cancers worldwide. Circular RNAs (circRNAs) play essential roles in the malignant progression and metastasis of gastric cancer. As a transcription factor, FOXP2 is involved in the progression of many tumours. However, the regulation and association between circRNAs and FOXP2 remain to be discovered. Methods: RNA sequencing was used to explore differential circRNA expression profile in gastric cancer and quantitative real-time PCR (qRT-PCR) were used to detect circST3GAL6 expression. The cellular location of circST3GAL6 was determined by fluorescence in situ hybridization (FISH). Functional experiments in circST3GAL6 knockdown and overexpression cell lines were performed in vitro and in vivo. The correlation between circST3GAL6 and miR-300 was confirmed by the RNA pull-down assay, dual-luciferase reporter assay and fluorescence in situ hybridization (FISH). The effects of circST3GAL6 on autophagy were detected by confocal microscopy and transmission electron microscopy (TEM). The mechanism of the circST3GAL6/miR-300/FOXP2 axis was verified by western blotting. The transcriptional regulation of Met by FOXP2 was proven by ChIP and luciferase reporter assays.Results: CircST3GAL6 was significantly depressed in GC tissues and cells. circST3GAL6 overexpression inhibited the proliferation, invasion and metastasis of GC cells in vitro and in vivo. Importantly, circST3GAL6 overexpression induced apoptosis and promote autophagy in GC cells. Furthermore, we found that circST3GAL6 sponged miR-300 and subsequently regulated FOXP2. We further revealed that FOXP2 suppressed the activation of the Met/AKT/mTOR axis, a classic pathway that regulates autophagy-mediated proliferation and migration.Conclusion: Our findings revealed that circST3GAL6 functions as a tumour suppressor through the miR-300/FOXP2 axis in GC, regulates apoptosis and autophagy through FOXP2-mediated transcriptional inhibition of the MET axis and may be a biomarker for GC treatment.


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Parvathi K. Basrur ◽  
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PLoS ONE ◽  
2019 ◽  
Vol 14 (8) ◽  
pp. e0221786 ◽  
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