The transcriptional activity of the Yes-associated protein 1 promoter in colon cancer cell lines

Nazu Salehi, Rikke Nørgaard Nielsen, Shukr Naser Karim Naser, Josephine Birkkjær Kristensen & Monika Scheuer Fløjborg

Student thesis: Subject Module Project

Abstract

Colorectal cancer (CRC) can occur due to a misregulation in the tumor suppressors,
oncogenes or pathways. Two pathways involved in CRC are the Hippo pathway and the Wnt
pathway. The components of the Hippo pathway interact with the Yes-associated Protein 1
(YAP1). The Caudal-type homeobox 2 (CDX2) protein, from the transcriptional regulatory
network, has shown to regulate the transcription of YAP1 in CRC cells. YAP1 and CDX2
normally regulate expression of genes in proliferation, cell adhesion, migration and
tumorigenesis. The purpose of this project is to investigate if the YAP1 promoter and
promoter with enhancer works in different colon cells. The intention is to use the Caco-2,
T84 and LS174T as cell models for colon cells, where the kidney cell line, HEK293, is used
as a negative control. For measuring the promoter activities with and without the enhancer
present, transfected into the cells, a promoter reporter assay is used to obtain the luciferase
and β-Galactosidase activities. For the examination of the YAP1 promoter activities, a Student
T-Test was performed to see whether there was a significant difference in the promoter
activities in the different cell lines. No significant difference was found between plasmids
containing the YAP1 promoter alone or those including both the YAP1 promoter and potential
enhancer. However, a tendency of transcriptional activity was seen in the expression of YAP1
in the different cell lines. Our results were compared with other studies that used human CRC
cell lines. The compared studies showed significant differences in the activities of the
promoter and enhancer within the different CRC cell lines and no significant difference in the
kidney cell line.

EducationsMolecular Biology, (Bachelor/Graduate Programme) Bachelor
LanguageEnglish
Publication date18 Dec 2018
Number of pages39
SupervisorsSylvester Larsen