Characterization of Glycosylation in a Cancer Model

Document Type

Article

Publication Date

Summer 2023

Keywords

JMG

JAX Location

In: Student Reports, Summer 2023, The Jackson Laboratory

Abstract

Cancer is diagnosed at high rates worldwide, which has prompted a shift towards approach towards individualized treatments as a variety of cancers are present in the population with unclear pathologies. Among molecular targets of interest for therapeutics are altered glycosylation patterns present on the surface of cancer cells. Glycosylation patterns mediate protein folding, cellular signaling, and migration processes, which appear altered in a cancerous state. The goal of this study is to determine the glycoprotein and phosphoprotein patterns of normal and cancerous epithelial cells in normal and high glucose environments, which will then be compared to a tissue-tumor model derived from the same cell types. To prepare samples for subsequent analysis via mass spectrometry, cell surface glycans were oxidized and biotinylated, cellular components were separated, proteins were digested, and peptides underwent glcyo- and phospho-enrichment. The work of this study so far has identified 30 phosphoproteins that have greater abundance in cancerous cells in a high glucose environment, as compared to a normal glucose environment. Initial glycosylation tests indicated the samples were too low in abundance, so that process is currently undergoing modification to increase detection. Once this continuing optimization work on cell surface glycosylation, cytosolic proteins, and tumor models is completed it will provide insight into the tumor microenvironment and the effect high glucose has on cancer proliferation.

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