| Objective:Glioma is a malignant tumor that originates from the glial cells of the brain and spinal cord.Gliomas are highly heterogeneous and aggressive.It is difficult to achieve radical resection by surgery,and the prognosis of patients is poor.Therefore,it will be imperative to explore the potential mechanisms involved in the development of glioma,as well as investigating the novel biomarkers for diagnosis and treatment targets.Preliminary bioinformatics analysis found that the survival time of patients with gliomas with high expression of miR-935 was significantly longer than that of patients with low expression of miR-935,and the expression of miR-935 in WHO grade IV gliomas was significantly lower than that of other grades of gliomas Mass tumor.Therefore,we speculated that miR-935 was down-regulated in gliomas and had an inhibitory effect on the occurrence and development of gliomas.To verify this conjecture,this study examined the expression of miR-935 in glioma tissues and cell lines,and analyzed the correlation between the expression of miR-935 and the WHO classification.The miR-935 overexpression glioma cell model was constructed,and in vivo and in vitro experiments were conducted to study its effects on the proliferation,invasion and migration of glioma cells and tumor formation in vivo.Methods: we collected clinical glioma tissues and normal tissues adjacent to tumors,and detected the expression of miR-935 in glioma tissues and normal tissues adjacent to tumors by real-time PCR;and then we compared the expression of miR-935 in TⅢ-TⅣ group and TⅠ-TⅡ group;We selected glioma cell lines(LN229,T98,U87,U251,U118,A172)and human normal astrocyte lines NHAS for culture,and detected the expression of miR-935 in glioma cell lines and in NHAS by RT-PCR;The overexpression miR-935 plasmid lentivirus was transfected into human glioma cell lines(T98,U87),and each group of cell lines was divided into two groups: the first group: the overexpression negative control group(NC);the second group: over expression miR-935 group(LV-miR-935);We used CCK-8 experiments,plate cloning experiments,and flow cytometry for cell cycle analysis to detect the proliferation ability of each group of cells;We used cell scratch test to detect the migration ability of each group of cells;We used the transwell invasion experiment to detect the invasion ability of each group of cells;We tested the tumorigenesis ability of each group of cells in vivo by subcutaneous tumor formation experiment in nude mice.Results: Compared with the normal tissues adjacent to the tumor,the expression of miR-935 in the glioma tissue was significantly down-regulated(P<0.05);compared with the TⅠ-TⅡ group,the miR-935 in the TⅢ-TⅣ group was significantly lower(P<0.01));Compared with normal human astrocytes(NHA),the expression of miR-935 in glioma cell lines T98,U87,U251,U118,A172,LN229 was significantly down-regulated(P<0.001);Compared with the NC group,the expression of miR-935 in LV-miR-935 group(U87,T98)was significantly up-regulated(P<0.01);In the CCK-8 experiment,plate cloning experiment,and cell cycle experiment,compared with the NC group,the cell proliferation ability of the LV-miR-935 group(U87,T98)was significantly inhibited,and the difference was statistically significant;In the cell scratch experiment,compared with the NC group,the cell migration ability of the LV-miR-935 group(U87,T98)was significantly inhibited,and the difference was statistically significant;In the Transwell cell invasion experiment,compared with the NC group,the cell invasion ability of the LV-miR-935 group(U87,T98)was significantly inhibited,and the difference was statistically significant;In the experiment of subcutaneous tumor formation in nude mice,the final volume and weight of the nude mice in the LV-miR-935 group were significantly lower than those in the NC group(P<0.01),and the body weight of the nude mice in the LV-miR-935 group was significantly higher than that in the NC group.NC group(P<0.01).Conclusions:Bioinformatic and q RT-PCR analyses revealed that the expression of miR-935 significantly decreased in glioma tissues in comparison with adjacent tissues.The expression of miR-935 in glioma tissues obtained from patients in Phase III and IV was also substantially lower than that of patients in Phase I and II.Lower expression levels of miR-935 in patients with glioma predicted reduced survival rate.Furthermore,overexpression of miR-935 significantly decreased proliferation,induced cell cycle arrest in glioma cells,and suppressed cell migration and invasion.These findings indicated that miR-935 can act as a suppressor in glioma and increased expression of miR-935 in glioma may inhibit its progression. |
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