MiR-330-5p Suppresses Glioblastoma Cellmalignant Phenotype Through Targeting ITGA5

Background Glioma is a common intracranial malignant tumor,originated from the central nervous system,about 45% of the central nervous system tumors.Morphologically,the glioma is highly invasive and has unclear boundaries.Therefore,it is difficult to completely remove the glioma and easy to relapse after surgery.Although surgery combined radiotherapy and chemotherapy has been the treatment guidelines,it is difficult to improve the prognosis of high grade glioma(WHO class Ⅲ,Ⅳ)patients by resection of the "largest range of safety" under image guiding and neural electrophysiological monitoring.Micro RNAs(mi RNAs,minute RNAs)are small,non-coding RNAs.In recent years,a large number of studies have been conducted on mi RNA.At present,it is known that it plays an important role in the occurrence and development of a variety of tumors,and has been considered as a new target of anti-tumor therapy in recent years.Most mi RNAs are involved in various human tumorigenesis and malignant phenotypes by directly targeting oncogenes or tumor suppressor genes.In this study,we studied a micro RNA,namely micro-330-5p,that has been less studied in glioma.The target gene for this micro RNA is ITGA5.However,the role of ITGA5 in different tumors is different,and its role in glioma is not yet clear.The purpose of this study was to investigate the functions and effects of mi RNA-330-5p and its target gene ITGA5 in glioma.This study provides a theoretical basis for mi RNA-330-5p to be used as a target for the diagnosis and treatment of glioma,and provides a new direction for the study of molecular regulation mechanism in the process of tumor progression.methods The expressions of mir-330-5p and ITGA5 m RNA in GBM cell lines(U87,U251 and U373)and normal glial cells(HEB)were detected by RT-q PCR.Western blot analyzesrelative protein expression.In this study,the biological functions(including cell proliferation,invasion,migration,apoptosis and cell cycle)of GBM cells were determined by MTT method,colony formation method,transwell method,wound healing method and flow cytometry.In this study,the relationship between mir-330-5p and ITGA5 was determined by double luciferase reporter gene detection.Tumor formation subcutaneously in nude mice was used to test the function of mi R-330-5p in vivo.Results RT-q PCR detect the expressions of mir-330-5p was lower in GBM cell lines(U87,U251 and U373)than that in normal glial cells(HEB).Western blot analysis indicated mi R-330-5p could decrease the expression of ITGA5.And then,it also alleviated the protein expression of Ki67,N-Cadherin and Vimentin,promote caspase-3 activation.In the biological function test,mir-330-5p up-regulation inhibited cell proliferation,invasion and migration,but promote the apoptosis of GBM cells.In vivo,U251 overexpressing mi R-330-5p was used to subcutaneous tumor formation in nude mice,which indicated a slower growth rate than NC group.Conclusion miRNA-330-5p can inhibit the expression of oncogene ITGA5,thereby inhibiting the proliferation,migration and invasion of glioma cells and promoting the apoptosis of tumor cells.Therefore,in-depth study on the mechanism of mir-330-5p inhibiting the malignant phenotype of glioma cells will provide a new direction for the clinical diagnosis and treatment of glioma,which has important clinical significance.