Meg3 Induces EMT And Invasion Of Glioma Cells Via Autophagy

Objective Glioma is one of the most common malignant tumors of the nervous system.Glioblastoma(grade ?) is considered the most malignant pathological form.At present,the treatment of glioma is mainly surgical combined with chemoradiotherapy.However,due to the invasion of glioma and its resistance to radiotherapy and chemotherapy,it is easy to relapse,greatly reducing the treatment effect.With the development of tumor genomics and molecular biology,more and more studies have confirmed the molecular biological mechanism related to the occurrence and development of glioma.Therefore,it is of great clinical significance to explore effective and feasible therapeutic targets to overcome the global problem of glioma.The relationship between long non-coding RNA Meg3 and tumor has been extensively studied.It has been reported that Meg3 is involved in the EMT process of liver and lung cancer cells.However,the relationship between Meg3 and EMT and invasion of glioma cells has not been fully investigated.In this study,we used recombinant plasmid transfection technology to overexpress the Meg3 gene in glioma cells,and then elaborated the effect of Meg3 on EMT and invasion of human glioma cells and its mechanism,so as to provide new ideas for the treatment of glioma.Methods(1)Real-time PCR was used to detect the expression abundance of Meg3 in primary cells isolated from tumor tissues of patients and human glioma cell lines.(2)Build Meg3 overexpressed cell model,cultivate Meg3 overexpressed glioma cells,observe the morphological changes under a microscope.The real-time PCR and Western blot were used to detect EMT related gene expression level after Meg3 overexpression in glioma cells.The influence of Meg3 overexpression on glioma cell migration and invasion ability was detected by Transwell migration and invasion assay.(3)Acridine orange staining and immunofluorescencethe were used to detect the effect of Meg3 overexpression on autophagy in glioma cells.Real-time PCR and Western blot were used to detect expression levels of autophagy related genes in glioma cells after Meg3 overexpression.(4)Western blot was used to detect the effectiveness of autophagy inhibitors on glioma cells and their effect on EMT of glioma cells.The microscope was used to observe the effect of autophagy inhibitors on glioma cell morphology.The effect of autophagy inhibitors on the expression of Meg3 gene was detected by real-time PCR in glioma cells.(5)Western blot and Transwell migration and invasion experiments were used to detect whether the effect of Meg3 overexpression on EMT,migration and invasion of glioma cells was affected by autophagy inhibitors.Result(1)Compared with primary cells isolated from tumor tissues and human glioma cell lines A172,the expression abundance of Meg3 gene in U87 and U251 glioma cell lines was lower.(2)Overexpression of Meg3 in U87 and U251 human glioma cell lines can induce EMT,migration,invasion and autophagy of human glioma cells.(3)autophagy inhibitors can effectively inhibit EMT of U87 and U251 glioma cells without affecting the expression of Meg3 gene.(4)In glioma cells,Meg3-induced EMT,migration,and invasion can be partially reversed by the autophagy inhibitors CQ(10?M)and Lys05(100?M).Conclusion All the experimental date suggested that Meg3 induced EMT and invasion of glioma cells via autophagy.Overall,the findings of the present study demonstrated the importance of Meg3 in the molecular etiology of glioma,which also indicated its potential applications in the treatment of glioma.