Microscopic analysis of aneuploidy induced by the mutation of the CCDC124 gene
Al-Murtadha, Asma Abdullah Abdulwahab (2015) Microscopic analysis of aneuploidy induced by the mutation of the CCDC124 gene. [Thesis]
The Coiled-coil domain containing protein 124 (Ccdc124) is a centrosomal protein that relocates to the midbody region at the cytokinesis stage of the cell cycle. Cytokinetic abscission is the cellular process that leads to physical separation of two postmitotic sister cells by severing the intercellular bridge. Mutation of the Ccdc124 gene by CRISPR/Cas9 genome editing in HEK293T cells leads to the failure of cytokinesis and formation of aneuploid (multinucleated-MN) aberrant cells. In this study, the MN cells were analyzed using flow cytometry and confocal imaging followed by quantitative image analysis. MN cells had mitotic and chromosome attachment aberrations, multiple centrosomes and micronuclei. These aberrations are known to occur in tumour cells, a finding that links Ccdc124 to cancer. MN cells also upregulated the p53 protein, which induced senescence. Furthermore, MN cells had increased numbers of 53BP1 foci which indicates that the mutation of Ccdc124 induces the DNA damage response and activates the p53 pathway. This study documents a relationship between Ccdc124 mutation-associated cytokinesis failure and p53-dependent senescence.
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