| Objective:The purpose of this study is to clarify the role of mi R-149-5p in glioma cells,and to explore whether MKI67 and MMP-9 are involved in the regulation of mi R-149-5p in glioma cells.Provide new directions for the treatment of glioma.Methods:q RT-PCR was employed to detect the expression of mi R-149-5p,MMP-9 and MKI67 gene in normal tissue and glioma tissue,Then we analysis the relationship between mi R-149-5p,MMP-9,MKI67 and clinical characteristics of glioma;Western Blot was employed to detect the protein expression of MMP-9 and MKI67 in normal brain tissue and glioma cells.The pathological tissue was collected within May 2010 to May 2020 from the neurosurgery department of the First Affiliated Hospital of Nanchang University,and divided into three groups according to the malignancy.First group: the low-grade glioma group—60 cases;second group: the high-grade glioma group—60 cases;third group: the recurrence glioma group—30cases.The fourth group: normal brain tissue group-30 cases,which was obtained from f patient with traumatic brain injury.Immunoassay was used to detect the content of MMP-9 and MKI67 in pathological tissue.At the same time,q RT-PCR was used to detect the mi RNA transfection efficiency in glioma cell lines;MTT assay was used to detect cell proliferation.Wound Scratch assay was employed to detect cell migration and Transwell assay was employed to detect cell invasion.t test was used to analyze the measurement data,and Pearson correlation analysis was used to test the correlation.When p<0.05,the difference was statistically significant.Results:We confirmed the existence of binding sites between mi R-149-5p and MMP-9,MKI67 gene by dual luciferase experiments..The positive expression rate of MKI67were: the low-grade glioma group(15.370±5.176)%;the high-grade glioma group(26.130±4.624)%;the recurrence glioma group(29.520±4.441)%;normal brain tissue group(0.000±0.000)%.The control group was compared with the resected tissue group of glioma patients,all p<0.05;the low-grade gliomat group was compared with the other two groups of glioma patients,p<0.05.The results of MMP-9 detection were:(28.030±3.943)%;(39.320±3.987)%;(41.230±5.377)%;and(1.245±0.390)%respectively.The comparison between the control group and the glioma group,bothp<0.05;the comparison between the first group and the second and third groups,the results are all p<0.05;the comparison between the second group and the third group,p>0.05.The examination results showed that the positive correlation between the positive rate of MKI67 and the positive rate of MMP-9 in glioma tissue was positive,p<0.05.And the expression of MKI67 and MMP-9 in human glioma was positively correlated with the malignant degree of glioma.Compared with normal brain cells,the expression of mi R-149-5p in human glioma cell lines U251,U87 and LN229 were significantly decreased(p<0.05);Western Blot experiment results showed that the expression of MMP-9 and MKI67 in human glioma cell lines U251 and U87 were significantly higher than those in normal cells(p<0.05).Compared with the mimic control group,the expression of MMP-9 and MKI67 in the mi R-149-5p mimic transfected glioma cells were significantly decreased(p<0.05).Compared with the mimic control group,the cell viability,cell migration and invasion ability of glioma cells in the mi R-149-5p mimic group were significantly decreased(p<0.05).Conclusion:The protein expressions of MMP-9 and MKI67,the target genes of mi R-149-5p,showed a gradually increasing trend in the percentage of positive cells.Overexpression of mi R-149-5p leads to a significant decrease in the expression levels of MKI67 and MMP-9,reducing the activity of glioma cells and their ability to migrate and invade,thereby inhibiting the occurrence and development of gliomas. |
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