Nestin Is Required for Spindle Assembly and Cell-Cycle Progression in Glioblastoma Cells

Nestin, a class IV intermediate filament protein, is generally considered as a putative marker of neural stem and progenitor cells in the central nervous system. Glioma is a common type of adult brain tumors, and glioblastoma (GBM) represents the most aggressive form of glioma. Here, we report that Nestin expression is significantly upregulated in human GBM, compared with other types of glioma. Nestin knockdown or deletion in U251 cells and tumor cells fromGBMpatients derived xenografts resulted in G(2)-M arrest, finally leading to apoptosis in tumor cells. Using proximity-dependent biotin identification method, we identified beta II-tubulin as an interacting protein of Nestin in U251 cells. Nestin stabilized beta II-tubulin in U251 cells through physical interaction. Knockdown of Nestin or beta II-tubulin disrupted spindle morphology in tumor cells. Our studies further revealed that Nestin deficiency in U251 cells and GBM PDX cells repressed tumor growth upon transplantation. Finally, we found that Nestin deficiency sensitized GBM cells to microtubule-destabilizing drugs such as vinblastine and vincristine. Our studies demonstrate the essential functions and underlying mechanisms of Nestin in the growth and drug response of GBM cells.