1. BUB1B is validated as a candidate glioblastoma-lethal gene in vitro.
BUB1B is validated as a candidate glioblastoma-lethal gene in vitro. A, BTIC-specific effects of BUB1B knockdown, visualized using shRNA–green fluorescent protein (GFP+) BTICs and NSCs 6 days after posttransduction with pGIPz-shRNA virus. Knockdown of KIF11/EG5, a microtubule motor protein critical for bipolar spindle formation during mitosis, was used as a positive control for both RNAi pathway activity and cell proliferation. B and C, examination of BUB1B knockdown by Western blot and quantitative real-time PCR (qRT-PCR) analysis in BTIC-G166 and NSC-CB660 cells. D, comparison of the effects of BUB1B knockdown on in vitro expansion of multiple BTIC and NSC lines and normal human astrocytes (NHA). **, Student t test; P < See Methods for a description of how BTIC isolates were developmentally subtyped. E, limiting dilution assays (LDA) for in vitro tumor sphere formation. BTIC-0131 cells and also unpassaged primary glioblastoma tumor cells (448T) were transduced with indicated LV-GFP-shRNAs, diluted, and assayed for sphere formation after 14 days. Linear regression analysis was used to evaluate significance.
Yu Ding et al. Cancer Discovery 2013;3:
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