The Protein Expression of CAV-1, COL1A2, and THBS2 proteins in the CaCo2 cell line.
Session Number
C29
Advisor(s)
Joel Pekow, University of Chicago
Location
A-115
Start Date
28-4-2016 1:35 PM
End Date
28-4-2016 2:00 PM
Abstract
Patients with Inflammatory Bowel Disease, which includes Crohn’s disease and ulcerative colitis (UC), are at a significantly higher risk than the general population for development of colon cancer. In preliminary studies, we identified down regulation of microRNA (miRNA), 4728-3p, in non-dysplastic mucosa from UC patients harboring a neoplastic lesion. Bioinformatics analysis suggests that this miRNA regulates three proteins also increased in patients with neoplasia in UC, and are vital to the interaction between cells and the extracellular environment. This investigation was performed to view caveolin 1, collagen, and thrombospondin 2 protein expression in the colon cancer cell line CaCo2, which is a human colorectal adenocarcinoma cell line. Before transfecting miRNA in CaCo2 cells, we measured baseline expression of the three proteins. Since the miRNA is predicted to down-regulate the proteins, high baseline expression is needed to demonstrate a difference. After culturing the CaCo2 cell line and running Western blots, it was concluded that the COL1A2 and THBS2 proteins were expressed, although CAV1 had low expression. Therefore, in subsequent experiments, we will use CaCo2 cells to examine the impact of the miRNA on COL1A2 and THBS2, and an alternative cell line to study the effects of miR-4728-3p on CAV1 expression.
The Protein Expression of CAV-1, COL1A2, and THBS2 proteins in the CaCo2 cell line.
A-115
Patients with Inflammatory Bowel Disease, which includes Crohn’s disease and ulcerative colitis (UC), are at a significantly higher risk than the general population for development of colon cancer. In preliminary studies, we identified down regulation of microRNA (miRNA), 4728-3p, in non-dysplastic mucosa from UC patients harboring a neoplastic lesion. Bioinformatics analysis suggests that this miRNA regulates three proteins also increased in patients with neoplasia in UC, and are vital to the interaction between cells and the extracellular environment. This investigation was performed to view caveolin 1, collagen, and thrombospondin 2 protein expression in the colon cancer cell line CaCo2, which is a human colorectal adenocarcinoma cell line. Before transfecting miRNA in CaCo2 cells, we measured baseline expression of the three proteins. Since the miRNA is predicted to down-regulate the proteins, high baseline expression is needed to demonstrate a difference. After culturing the CaCo2 cell line and running Western blots, it was concluded that the COL1A2 and THBS2 proteins were expressed, although CAV1 had low expression. Therefore, in subsequent experiments, we will use CaCo2 cells to examine the impact of the miRNA on COL1A2 and THBS2, and an alternative cell line to study the effects of miR-4728-3p on CAV1 expression.