Here, we found that MLK3 mRNA levels in human gliomas was negatively related to EGFR and MAPK8, MAPK9, MAPK10 mRNAs levels, and MLK3 downregulation reduces GBM cell migration and invasion without EGF induction, suggesting that the MLK3 regulation of cell migration and invasion is EGF signaling-independent. This evidence concerns the gene EGFR and central nervous system cancer.