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The Soft Agar Colony Formation Assay
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CircFBXW7 alleviates glioma progression through regulating miR-23a-3p/PTEN axis
Zhi-Gang Gao1, Peng Yang1, Jie Huang1
1Department of Neurosurgery, People's Hospital of Lanling County, Linyi, Shandong, China.
Abstract:
Increasing evidence has confirmed that circular RNAs (circRNAs) are involved in regulating the development and progression of various tumors. The aim of this study was to examine the effect of circFBXW7 on the progression of glioma and to determine its underlying mechanism. qRT-PCR was performed to measure the expression of circFBXW7, miR-23a-3p, and PTEN in tissues and cell lines of glioma. The proliferation ability of glioma cells was examined using the CCK-8 assay. Glioma cell migration and invasion capacity were detected using Transwell assays. The dual-luciferase reporter gene assay was employed to examine the correlation between miR-23a-3p and circFBXW7 or PTEN. The expression levels of the related genes were determined using western blotting analysis. A glioma xenograft tumor model was employed to evaluate the functional roles of circFBXW7 in vivo. CircFBXW7 was found to be aberrantly downregulated in glioma tumor tissues and cell lines. Overexpression of circFBXW7 was found to significantly inhibit the proliferation, migration and invasion ability of the glioma cells. Moreover, bioinformatic analysis and dual-luciferase reporter assays confirmed that circFBXW7 can directly target miR-23a-3p, which then blocks the binding of miR-23a-3p to the 3' un-translated region (UTR) of PTEN. Mechanically, circFBXW7 suppresses cell proliferation and metastasis in glioma by sponging miR-23a-3p, resulting in elevated PTEN expression. In addition, in vivo experiments also confirmed that circFBXW7 overexpression effectively halts tumor growth and metastasis. Consistent with the in vitro observations, circFBXW7 overexpression significantly decreased miR-23a-3p, Ki-67, and N-cadherin, as well as increased PTEN and E-cadherin levels. Our results revealed that circFBXW7 exhibits antiproliferative and antimetastasis activities via sponging miR-23a-3p to elevate PTEN expression in glioma, which may offer a novel target for clinical therapy and diagnosis of glioma.
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