Tao Lu
Cancer Center Indiana University School of Medicine, Indianapolis
Title: VBIM technology identifies adenomatous polyposis coli like protein (ALP) as a novel negative regulator of NF-ï«B
Biography
Biography: Tao Lu
Abstract
Colon cancer is the second leading cause of cancer related deaths in the United States. The nuclear factor ï«B (NF-ï«B) is an important family of transcription factors whose aberrant activation has been found in many types of cancer, including colon cancer. Therefore, understanding the regulation of NF-ï«B is of ultimate importance for cancer therapy. The purpose of this study is to use a novel validation-based insertional mutagenesis (VBIM) strategy to identify novel regulators of NF-ï«B, and further evaluate their roles in the regulation of NF-ï«B signaling in colon cancer cells. We infected Z cells (293 derived cells with hyper active NF-ï«B activity) with VBIM virus to cause the overexpression of negative regulators of NF-ï«B, and then further selected the mutant cells with low NF-ï«B activity under ganciclovir (GCV) treatment. Targeted gene was then identified by using VBIM specific primers. In a preliminary screen, we identified the novel adenomatous polyposis coli like protein (ALP) gene as a negative regulator of NF-ï«B. Overexpression of ALP led to decreased NF-ï«B activity by ï«B reporter assay, while knocking it down had the opposite effect. Furthermore, we found that overexpression of ALP in HT29 colon cancer cells greatly reduced both the number and the size of colonies that were formed in a soft agar assay, while using shRNA resulted in an opposite effect, confirming that ALP is a tumor suppressor in HT29 cells. Future experiments aim to further assess the role of ALP in colon tumor formation in a mouse xenograft model. In summary, by using the novel VBIM technique, we identified ALP as a novel negative regulator of NF-ï«B. This discovery could lead to the establishment of ALP as a potential biomarker and therapeutic target in colon cancer.