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The miR-192-EGR1/HOXB9 Loop Regulates Glioma Cell Stemness and Malignant Phenotypes by Promoting Their Mesenchymal
Guo-Wei Li1, Yan-Ping Jin2, Min-Feng Sheng3
1School of Rehabilitation Science, Nanjing Normal University of Special Education, Nanjing, Jiangsu, China.
Abstract:
To clarify the regulatory effects of miR-192 on the malignant phenotypes of glioma cells. We used PCR, WB and immunofluorescence to detect regulatory factors in glioma samples. Then, we chose lentiviral plasmid transfection to construct cell models. We used CCK-8 and colony formation to evaluate the proliferation ability of these cells and used Transwell/scratch tests to evaluate their invasion ability. CD133-expressing GSCs were observed under a microscope, and their stemness properties were evaluated. We constructed a tumour-bearing model via subcutaneous inoculation. Tumour growth curves and tumour weights were determined subsequently. The proteins involved in the miR-192-EGR1/HOXB9 loop were evaluated via IHC staining. MiR-192 was significantly reduced in glioma samples, and this factor downregulated EGR1 and HOXB9 via targeted binding, thus forming a semi-open loop. Moreover, the proliferation, invasion and migration of glioma cells overexpressing miR-192 were significantly decreased. These malignant phenotypes were abrogated completely with EGR1 or HOXB9 overexpression. Similarly, these changes were essentially consistent with MT marker expression and the stem-like properties in glioma cells. Meanwhile, miR-192 inhibits the tumorigenesis of glioma cells via the EGR1-HOXB9 loop. MiR-192 could inhibit MT in glioma cells through the EGR1-HOXB9 loop. Thus, it reduces their stemness and abrogates their malignant phenotypes.
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