Which Drug Inhibits H3K27me3 ​Better in Human Melanoma Cells?​

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Chandler Ross

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Poster presented at the Annual Biomedical Research Conference for Minoritized Scientists (ABRCMS) in Pittsburgh, PA.

Metastatic cancer cells move through tight junctions smaller than their own nucleus, in order to reach a secondary site. Exerting force on the cell's nucleus can often cause changes in the 3D genome structure, gene transcription, and the addition of epigenetic marks. Previous findings have classified A375 cells that have undergone ten rounds of constricted migration as Bottom-10 cells and those that have not as Parental cells. During constricted migration, the A375 cell line is marked with H3K27me3. Previous data suggests that this histone is a major component of the chromatin compartment switches and gene expression of Bottom-10 cells following constricted migration. H3K27me3 is more expressed in Bottom-10 cells in downregulated gene expression and during shifts from the active (euchromatin) to inactive (heterochromatin) compartment. Within this project, we aim to understand the impact of H3K27me3 on chromatin compartment switches and transcriptional differences between parental and Bottom-10 A375 cells through the use of an H3K27me3-specific inhibitor. The enhancer of zeste homolog 2 (EZH2), which is responsible for the methylation of H3K27me3, was inhibited using the drugs UNC1999 and EPZ005687. During constricted migration, histones typically lose acetylation marks and gain methylation marks to prevent transcription. However, the inhibition of EZH2 causes the removal of methylation marks to allow transcription. To analyze the impact of the epigenetic mark, H3K27me3, following its inhibition, immunofluorescent staining and imaging procedures were used to identify H3K27me3 signals within the cells.

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