Effects of Transcription Factors Associated with Colorectal Cancer on SOX9 Promoter Activity in Caco-2 Cells: Bachelor Project

Alina Malita, Mette Lassen, Eva Rose Jepsen, von Heymann-Horan & Kevin Marcellus

Studenteropgave: Semesterprojekt

Abstrakt

The purpose of this study was to investigate the regulation of SOX9 in colon cancer. Although SOX9 is significant in organ development and patterning, little is known about its function in colorectal cancer. Moreover, SOX9 regulation by transcription factors which have been found to play a role in colorectal cancer is almost non-existent, with the exception of some Wnt pathway components. Therefore, an investigation of the SOX9 promoter activity was conducted by transfecting Caco-2 cells with a pGL4.10 vector containing a luciferase gene under the control of the SOX9 promoter. Clones of the SOX9 plasmid from two different bacterial colonies were used separately in two sets of transfection. Sequence analysis of the two clones and alignment with the original promoter sequence showed no significant differences (data not shown). Additional co-transfections contained other plasmids expressing transcription factors correlated with colon cancer: CDX2, TCF4, HNF4a, Sp-1, c-jun, c-fos, p65, p52, and p50. All samples were analyzed with a luciferase assay to determine the SOX9 promoter activity under the influence of the aforementioned transcription factors. The transcription factors P65 + P52, CDX2 + HNF4-α, and HNF4-α increase the promoter activity significantly, determined by a Student T-Test. P65 + P50, c-jun + c-fos, and SOX9 decrease the SOX9 promoter activity. In the one clone Sp1 appears to increase the promoter activity while in the other clone it decreased promoter activity. A 2-tailed independent sample Student T-Test was used to check for reproducibility between the two clones, and it was found that all samples except the CDX2 + HNF4-α, Sp1, and P65 + P52 were reproducible.

UddannelserMolekylærbiologi, (Bachelor/kandidatuddannelse) Bachelor el. kandidat
SprogEngelsk
Udgivelsesdato19 jun. 2015
VejledereMarcker Espersen & Maiken Lise

Emneord

  • CaCo-2
  • CDX2
  • Colorectal cancer
  • pGL4.10
  • Luciferase assay
  • c-fos
  • P50
  • P52
  • P65
  • c-jun
  • HNF4-α
  • SP1
  • TCF4
  • SOX9