LabAutomation 2006 - SLAS

MP37
Weisong Gu
Ohio Supercomputer Center
Springfield, Ohio
weisong@osc.edu
Where Laboratory Technologies Emerge and Merge
Co-Author(s)
Xi Chen
Paul Evans
University of Texas
Eric Stahlberg
Ohio Supercomputer Center
Chunming Liu
University of Texas
Genome-Wide Location and Analysis of b-catenin/TCF Target Genes
Wnt/b-catenin signaling plays essential roles in both development and tumorigenesis. Wnt signaling is mediated by b-catenin, which binds
T cell factor (TCF) in the nucleus and activates gene transcription. In the absence of Wnt stimulation, a protein complex consisting of
Glycogen synthase kinase-3 (GSK-3), Casein kinase I alpha (CKIa) and tumor suppressor proteins Axin and Adenomatous polyposis coli
(APC), phosphorylates b-catenin. The phosphorylated b-catenin is degraded by the ubiquitin/proteasome pathway. However, mutations in
the Wnt/b-catenin signaling pathway prevent b-catenin degradation. Accumulated b-catenin enters the nucleus and forms a complex with
TCF and activates TCF target genes that ultimately lead to tumor formation, e.g. colorectal cancers. Although gene expression microarray
studies have revealed some b-catenin/TCF related genes, many of them are actually not regulated by b-catenin/TCF directly. To identify
the complete direct target genes that b-catenin/TCF transcribes, a custom human promoter array has been designed locating all possible
candidate TCF binding sites throughout the human genome. ChIP-on-chip analysis is performed in human colon cancer cell lines using
antibody against TCF4. A high resolution map of b-catenin/TCF target genes is constructed using Bioinformatics approach, and the genetic
b-catenin/TCF regulatory network is further characterized based on data from both gene expression microarray and ChIP-on-chip assay.
MP38
Kurtis Guggenheimer
University Of British Columbia
Vancouver, British Columbia, Canada
kurtis@physics.ubc.ca
Co-Author(s)
Jared Slobodan
Mark Homenuke
Keddie Brown
Roy Belak
Andre Marziali
Automation of Novel Protocols for Immunohistochemistry Staining
In an effort to reduce costs associated with cancer diagnosis, a need has been realized for a clinical device that can automate
immunohistochemical staining of individual cell biopsies located on a tissue microarray. This device, named the Cancer Biopsy Array
Spotter, or CBAS, is currently being developed and will provide fundamental improvements over current biopsy analysis techniques.
Precision application of costly primary antibody solutions to each individual biopsy will reduce the volume of solution needed, and therefore,
lower the analysis costs associated with purchasing these reagents. In addition, the CBAS will allow the use of many different antibody
solutions on one microarray, as opposed to the current method, which involves batch treatment of the whole slide. This feature will allow
one to test for many different forms of cancer simultaneously. A novel method for delivering and incubating nanolitre volumes of reagent
has been designed and developed for the CBAS. An overview of the CBAS and novel immunohistochemistry staining protocols will be
presented.
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