CROCC abrogates centrosome-related mitotic errors in 1p36

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CROCC abrogates centrosome-related mitotic errors in 1p36 CROCC abrogates centrosome-related mitotic errors in 1p36.13-deleted cancer cells. CROCC abrogates centrosome-related mitotic errors in 1p36.13-deleted cancer cells. A, Images of T84 cells with a large micronucleus (white arrow) and significantly reduced CROCC staining (enlarged in insets), anaphase bridges (arrow), or monopolar spindle (arrow) associated to deficient or fragmented γ-tubulin dots (enlarged in insets). Scale bar, 5 μm. The top right graph reports the percentage of anaphase showing segregation errors and micronuclei in T84 cells with serum supplementation or serum deprivation (SD) at 12 hours. Error bars represent mean ± SEM; *, P < 0.05, two-tailed Student t test). B, Representative images of T84 cells transfected with full-length human CROCC-GFP or GFP alone, immunostained for γ-tubulin (enlarged in insets). The graph on the right shows the survival of T84 transfected with CROCC and matched control cells maintained in neomycin selection (0.6 μg/mL) for the indicated time. Viability was assessed by a colony formation assay. The GFP vector was used as a control. Cells were fixed, stained, and photographed after 6 and 12 days of culture. C, The left graph reports flow cytometry analysis after 6 days. Error bars represent mean ± SEM of five independent experiments; **, P < 0.01; ***, P < 0.001, two-tailed Student t test). Tetraploid on diploid cells ratio after 6 days quantified by metaphases spreads (16 independent experiments for each condition; **, P < 0.01, two-tailed Student t test). The right graph shows the percentage of γH2AX foci in prometaphase (>250 cells per cell line, ***, P < 0.001, Mann–Whitney U test). D, Schematic representation of rhabdoid colorectal cancer progression. In colorectal cancer with defective microtubule functions, BRAFV600E mutation depletion of CROCC causes defective centrosome structure, abnormal mitotic progression, and lethal cancer phenotypes. Andrea Remo et al. Mol Cancer Res 2018;16:1385-1395 ©2018 by American Association for Cancer Research