Microarray profiling analysis of microRNAs expression in glioblastoma cells

Although growing evidence has revealed that ZEB2 was correlated with grades and outcome of glioma positively, the mechanisms underlying ZEB2/miRNAs-mediated gliomas development remain largely unknown. Here the authors revealed that ZEB2 raised in glioblastoma samples compared to the lower-grade gliomas. Besides, miR-637 increased to the most extent per the miRNA array analysis of cells with- or without ZEB2. ZEB2 captured the promoter of miR-637 directly and suppressed miR-637 transcription. In the absence of ZEB2, overexpressed miR-637 targeted WNT7A and repressed the WNT/β-Catenin signaling pathways subsequently, which inhibited the expression of Cyclin D1 and Vimentin. Importantly, the combination of β-Catenin-specific inhibitor LGK974 and temozolomide hampered cell proliferation synergistically in cells expressing miR-637. In line with the expectation, the inhibition of miR-637 counteracted sh-ZEB2-caused tumor regression in U251 xenograft models. The current study evaluated a novel mechanism that the association of ZEB2/miR-637/WNT7A contributed to regulate glioblastoma cell proliferation and migration in a β-Catenin-dependent manner. The results may provide new rationales to optimize the trial designs, which try to select the ideal glioblastoma patients to receive β-Catenin-targeted therapy and chemotherapy. Three U251 sh-NC clones and three U251 sh-ZEB2 clones were generated in the present study. Microarray based miRNA expression profiles were obtained with miRCURY LNA miRNA custom probe PCR assays.