MiR-129-5p inhibits the proliferation, migration, and invasion of U87 glioblastoma cells by targeting CaMK IV via the MAPK signaling pathway

Abstract

Objective The purpose was to investigate the expression of CaMK IV in glioblastoma (GBM) and its specific role in GBM cell proliferation, invasion, migration, and other malignant phenotypes. Furthermore, this study aimed to validate the functional relationship between miR-129-5p and CaMK IV, then explore the underlying molecular mechanisms involved. Results Bioinformatics analysis revealed that CaMK IV was expressed at higher levels in normal brain tissues than in GBM tissues. qRT-PCR demonstrated significantly lower expression of CaMK IV in GBM cell samples than in the control tissue. CCK-8 assays indicated that overexpression of CaMK IV significantly inhibited the activity and proliferation of U87 GBM cells, while knockdown of CaMK IV had the opposite effect. Similarly, the introduction of miR-129-5p mimics inhibited the activity and proliferation of U87 GBM cells, while the use of a miR-129-5p inhibitor yielded the opposite results. The scratch assay revealed that, compared with control treatment, CaMK IV overexpression led to larger scratch areas at 12 and 24 hours, indicating that CaMK IV has an inhibitory effect on the migration ability of the GBM cell line U87. Conversely, knockdown of CaMK IV had the opposite effect. Moreover, Transwell assays demonstrated that the overexpression of CaMK IV resulted in significantly fewer penetrating cells than was observed in the control group, suggesting that CaMK IV has an inhibitory effect on the invasion ability of U87 GBM cells. Again, knockdown of CaMK IV yielded the opposite results. Western blot analysis revealed that the overexpression of CaMK IV significantly suppressed the expression of proliferation-related proteins while promoting the activity of apoptosis-related proteins in the U87 GBM cell line. Moreover, overexpression of CaMK IV inhibited the phosphorylation of the MAPK pathway-related proteins JNK and P38, indicating CaMK IV has an inhibitory effect on GBM cells via the MAPK signaling pathway. Conclusion MiR-129-5p selectively inhibits the proliferation, migration, and invasion of U87 cells by targeting CaMK IV via the MAPK signaling pathway. MiR-129-5p and CaMK IV play an important inhibitory roles in GBM and may serve as potential therapeutic targets for GBM treatment.