University of Texas MD Anderson Cancer Center (UT-MDACC): High-throughput Screening Identifying Driving Mutations in Endometrial Cancer
Recent advances in next-generation sequencing technology have enabled the unprecedented characterization of a full spectrum of somatic alterations in cancer genomes. Given the large numbers of somatic mutations typically detected by this approach, a key challenge in the downstream analysis is to distinguish “drivers” that functionally contribute to tumorigenesis from “passengers” that occur as the consequence of genomic instability. For this study, the CTD2 Center at the University of Texas MD Anderson Cancer Center implemented a systems biology approach to identify driver cancer genes in endometrial cancers.
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Whole exome sequencing was performed on 13 endometrial cancers along with matched normal samples to detect somatic alterations with high precision and sensitivity. For validated mutated genes, bioinformatics prioritization was combined with high throughput screening of shRNA and overexpression constructs in a highly sensitive BaF3 cell viability assay to identify candidate driver cancer genes. The BaF3 cell line is a murine bone marrow-derived pro-B-cell line that depends on interleukin-3 (IL-3) for proliferation, but readily becomes IL3-independent in the presence of an oncogene or oncogenic event.
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