WNT-β-Catenin Is Upregulated in Very Aggressive Tumor Pattern of Invasion
Tatyana Isayeva, Margaret Brandwein-Gensler. UAB, Birmingham, AL
Background: The WNT pathway is a critical to proliferation, differentiation, oncogenesis and is upregulated in many cancers. Among the downstream effects, WNT-β-catenin is involved in tumor invasion. We developed 4 cell lines from oral/oropharyngeal SCC, which are unique as they are matched to different worst patterns of invasion (WPOI) in the corresponding resections. These histopathological WPOI were developed and validated by us as outcome prognosticators.
Design: Cell lines were derived from fresh resections, cancer cells were sorted from cancer-associated fibroblasts by flow cytometry and grown in keratinocyte media/keratinocyte growth supplements. HPV was determined by RT-PCR with HPV16/18 E6/E7 primers, with appropriate controls. Invasion was measured with the BioCoat Matrigel chamber, 24h after stimulation with TGFß. Cells that invaded across the chamber membrane were counted after fixation and staining. UAB-1, UAB-4, and the corresponding frozen samples from the 2 resections were assessed for WNT signalling with the RT2 96 well RT-PCR kit (SABiosciences).
Results: Table 1 shows the clinicopathologic features for these cells.
|Pattern of invasion, corresponding resection||WPOI3||WPOI3||WPOI5||WPOI5|
|Gene||Tumor-1 (WPOI3)||UAB-1 derived from Tumor-1||Tumor-2 (WPOI5)||UAB-4 derived from Tumor-2|