Culture based screening assay and methods of use thereof to identify agents which modulate tumor development, invasion and differentiation

摘要

A 3D organotypic culture which phenocopies aggressive, invasive cancer and methods of use thereof are provided.

主权项

1. A method for in vitro screening of a compound comprising:
a) providing an organotypic culture comprising an artificial stroma of collagen and human fetal esophageal fibroblasts, said fibroblasts comprising an inducible AKT gene, and being overlayed with
i) esophageal keratinocytes transduced with at least one activated oncogene and a transduced nucleic acid which inhibits p120ctn action selected from the group consisting of small hairpin RNA, siRNA and microRNA, orii) keratinocytes present in an esophageal tumor isolated from a patient; b) contacting said cultures with said compound and an AKT inducing agent, AKT induction producing a more invasive, less differentiated cancer phenotype; c) observing the effect of said compound upon migration and invasive characteristics of said transduced esophageal keratinocytes or esophageal tumor keratinocytes; and d) determining whether said compound modulates expression levels of periostin.