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- W2912640447 abstract "Cancer cells exhibit genomic instability, with gains or losses of entire chromosomes and chromosome segments that cause massive changes in gene expression. This aneuploidy occurs early in cancer development and is particularly prominent, ongoing, and variable in solid tumors – stiff tumors in particular. Here we develop live cell fluorescence monitoring for loss or gain of two chromosomes (Ch3 (green) and Ch5 (red)) in a lung cancer-derived cell line in order to quantify factors that affect aneuploidy. In standard culture, ∼10% of mitotic cancer cells show chromosome mis-segregation, but mis-segregation increases several-fold with mechanical pressing during mitosis or drug inhibition of the spindle assembly checkpoint. Because multiple DNA repair factors mis-localize transiently from nucleus to cytoplasm when cells exhibit nuclear envelope rupture, transient knockdown of such physically-regulated factors was also studied, with knockdown cells showing more DNA damage and more chromosome mis-segregation. After such perturbations, more red-fluorescent-null cells are observed indicating a loss of Ch5. The aneuploid changes are confirmed by single cell karyotyping as well as SNP arrays and marker-specific PCR on stable clones. Single cell transcriptomics of these red-null cells further reveal gene expression on Ch5 is significantly downregulated in >80% of cells. In addition, ∼10% total genes on average are mis-regulated more than 2-fold in response to such Ch5 loss. Normal cells are being similarly studied, but viability seems much lower with chromosome loss. Such system thus lend insight into factors affecting genome variation, gene expression, and cell viability." @default.
- W2912640447 created "2019-02-21" @default.
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- W2912640447 date "2019-02-01" @default.
- W2912640447 modified "2023-10-17" @default.
- W2912640447 title "Loss of Chromosomes Monitored in Live Cells" @default.
- W2912640447 doi "https://doi.org/10.1016/j.bpj.2018.11.2238" @default.
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