BCL-2's effect on gene regulation in MCF10A cells as detected by qRT-PCR / by Andrew R. Taylor.

Author/creator Taylor, Andrew R. author.
Other author McConnell, Thomas J., degree supervisor.
Other author East Carolina University. Department of Biology.
Format Theses and dissertations
Production2009.
Descriptionviii, 45 leaves : illustrations (some color) ; 28 cm
Supplemental ContentAccess via ScholarShip
Subjects

Summary Cells over-expressing the proto-oncogene B-cell lymphoma (BCL2), belonging to the Bcl-2 family of proteins, have been shown to slow the cell cycle and resist apoptosis. Microarray data has shown differences in a number of genes within and between several gene systems in the MCFI0A cell line. We hypothesized that genes shown to be differentially expressed in MCFI0A cells could be further verified through quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR). Using Ingenuity Pathway Analysis, twelve differentially expressed genes were chosen from the top five highest rated networks of genes from the microarray data. Through the use of the comparative C1 method ten of the genes were found to be up-regulated, while the remaining two were shown to be down-regulated. The two genes shown to be down-regulated were MCL-1 and FOXMl. The ten genes shown to be up-regulated are as follows: THRA, ITGB5, MAPIIIKII, ANXAl, PSENEN, RPS6KB1, HCLSl, PYCARD, RPS6KB2, and USP3. Of the twelve genes, seven correlated with the microarray data and are: MCLl, PSENEN, RP6KB2, HCLSl, PYCARD, and USP3. One-way analysis of variance (ANOVA), a statistical test, showed that the differential expression of ANXAl, PSENEN, HCLSl, and PYCARD were significant. We concluded that the over-expression of BCL2 does affect the expression of other genes within the MCFI0A cell line, and that these expressional changes could be directly affecting the regulation of apoptosis and the cell cycle. The exact mechanism to how this regulation is occurring remains unclear, however these experiments have provided a better understanding of gene expression regulation in MCFI0A cells.
General notePresented to the faculty of the Department of Biology.
General noteAdvisor: Thomas J. McConnell
Dissertation noteM.S. East Carolina University 2009
Bibliography noteIncludes bibliographical references (leaves 38-45).
Genre/formdissertations.
Genre/formAcademic theses.
Genre/formAcademic theses.
Genre/formThèses et écrits académiques.

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