The Effect Of MicroRNA-195in Capability Of Migration And Invasion Of Human Glioma Cell Line

Objected:To explore how the expression level of miR-195is related to the migration and invasion of U251and SHG-44, and identify the possible mechanism of this phenomenon.Method:Using We oligonucleotides miR-195mimics transfection of human glioma cell line SHG44, U251, and Real-time PCR detection of the expression of miR-195, and then through the scar healing assay cell migration, take photos of which the image was processed via J image processing software to calculate the migration area Oh and24hafter scratched; using the Transwell method detect glioma cell migration and invasion. The experimental data obtained were expressed as mean+standard.SPSS17.0software was used for statistical analysis of the experimental data. One-way ANOVA was used in the comparison of several groups’ mean, and LSD-t test was used to analyses the variance among the groups in the number of pairwise comparisons.P<0.05was defined as statistically significant.Result:Transfection efficiency observed under a fluorescence microscope was more than90%, the qRT-PCR melting curve shows the melting curves are single peak of miR-195, as well as internal reference U6, showing specific primers, a single product; amplification curve shows obvious logarithmic amplification of a typical PCR amplification curve, the number of amplification and plateau, indicating that there is enough PCR product this shows primer has higher sensitivity; PCR results showed that for the untreated cell lines blank control, miR-195expression levels of miR-195 transfection group were significantly increased,6to7times higher of the control group; cell wound healing experiment shows blank and control groups in the48hours after a large number of cells has been moved to the center of the scratches, the miR-195transfected group, the number of invasion cells was significantly less than the first two groups; the number of cells migrated Transwell experiments in U251cell line blank control groups and miR-195transfection group (416.9±28.7)(469.2±27.6)(221.3±28.3); blank group, control group and miR-195transfected cells group migration in Transwell experiments of SHG-44cell line is (264.4±26.9)(251.9±25.9)(397.2±37.9),The difference was statistically significant.Conclusion:The suppressive effect of miR-195mimics group is better than control and blank group in U251vitro,which indicate that miR-195may function as tumor suppressor in human gliomas. miR-195acting as a potent target of gene therapy with bright future.But in SHG-44vitro, miR-195function as a tumor gene improves the migration ability of cells.