Effect Of Tata-Box-Binding-Protein Associated Factor 1 (Taf1) And P0071 (An Adherens Junction Protein) On Cell Polarity Of Caco-2, A Human Colon Carcinoma Cell Line

Zachary Berry

School Name

South Carolina Governor's School for Science and Mathematics

Grade Level

12th Grade

Presentation Topic

Cell and Molecular Biology

Presentation Type

Mentored

Mentor

Mentor: Ilse Hofmann, Division of Vascular Oncology and Metastasis, German Cancer Research Center (DKFZ

Abstract

Cell polarity plays a vital role in cell functionality. In human carcinomas, epithelial cell polarity is destroyed, causing cells to divide and migrate rapidly. From a genome-wide siRNA scan in Drosophila ovaries, the downregulation of the transcription factor TATA-box-binding-protein associated factor 1 (TAF1) showed a major decrease in the polarity of the very clear apical-basal markings of the lining of the Drosophila ovaries. The aim of this research was to explore the effects of TAF1 along with p0071, an adherens junction protein, on the polarity of Caco-2 cells, a human colon carcinoma cell line. The TAF1 and p0071 proteins were then downregulated through reverse siRNA transfection to see the effects on the cell junctions: adherens junctions, tight junctions, and desmosomes. Adherens junctions hold epithelial cells together while assisting in contact inhibition. Tight junctions make small, impermeable channels between cells and desmosomes help to connect cells, but they also assist in attaching to the basal lamina. The knockdowns of the two proteins were checked by Western Blot analysis. The immunofluorescence technique was then used along with microscopy to view the co-localization of TAF1 and p0071 with E-cadherin (adherens junction protein), Cingulin (tight junction protein), and Desmoplakin (desmosome protein). The 2D culture of immunofluorescence showed that there was no effect of the TAF1 and p0071 downregulation on cell junctions. However, since colon cells normally form tubes that attach to the basal lamina, the downregulation of TAF1 in a 3D set-up might disrupt the polarity of the colon cell line.

Start Date

4-11-2015 9:00 AM

End Date

4-11-2015 9:15 AM

COinS
 
Apr 11th, 9:00 AM Apr 11th, 9:15 AM

Effect Of Tata-Box-Binding-Protein Associated Factor 1 (Taf1) And P0071 (An Adherens Junction Protein) On Cell Polarity Of Caco-2, A Human Colon Carcinoma Cell Line

Cell polarity plays a vital role in cell functionality. In human carcinomas, epithelial cell polarity is destroyed, causing cells to divide and migrate rapidly. From a genome-wide siRNA scan in Drosophila ovaries, the downregulation of the transcription factor TATA-box-binding-protein associated factor 1 (TAF1) showed a major decrease in the polarity of the very clear apical-basal markings of the lining of the Drosophila ovaries. The aim of this research was to explore the effects of TAF1 along with p0071, an adherens junction protein, on the polarity of Caco-2 cells, a human colon carcinoma cell line. The TAF1 and p0071 proteins were then downregulated through reverse siRNA transfection to see the effects on the cell junctions: adherens junctions, tight junctions, and desmosomes. Adherens junctions hold epithelial cells together while assisting in contact inhibition. Tight junctions make small, impermeable channels between cells and desmosomes help to connect cells, but they also assist in attaching to the basal lamina. The knockdowns of the two proteins were checked by Western Blot analysis. The immunofluorescence technique was then used along with microscopy to view the co-localization of TAF1 and p0071 with E-cadherin (adherens junction protein), Cingulin (tight junction protein), and Desmoplakin (desmosome protein). The 2D culture of immunofluorescence showed that there was no effect of the TAF1 and p0071 downregulation on cell junctions. However, since colon cells normally form tubes that attach to the basal lamina, the downregulation of TAF1 in a 3D set-up might disrupt the polarity of the colon cell line.