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Possible involvement of NF-?B in the response of cells to folate deficiency.

UNCG Author/Contributor (non-UNCG co-authors, if there are any, appear on document)
Rashmi Ranjana (Creator)
Institution
The University of North Carolina at Greensboro (UNCG )
Web Site: http://library.uncg.edu/
Advisor
Karen Katula

Abstract: Folate, a water soluble B vitamin, serves as a methyl donor for the synthesis of nucleotides and methionine. Folate deficiency has been linked to a variety of health problems that include developmental defects, cancer and neurological diseases. The relation between folate deficiency and human health problems has not been fully explained. Our preliminary results and limited published data have shown that the activity of NF-? B increases in folate deficient conditions. NF-?B is a cytosolic dimeric transcription factor that responds to a variety of stimuli by regulating genes involved in immune response, cell survival and cell growth. The purpose of the study was to determine if folate deficiency leads to altered NF-?B activity, protein levels and nuclear localization, and to determine the possible mechanisms behind this alteration. NIH3T3 cells containing a NF-?B reporter construct (NF-?B-LUC-3T3 cells) were grown for 7 and 11 days in folate sufficient and folate deficient DMEM medium either with or without TNF-a (stimulator of NF-?B). NF-?B activity was higher in folate deficient medium at days 7 and 11 in both unstimulated and stimulated cells, as measured by luciferase assay per unit DNA. NF-?B protein levels analyzed by Western Blot showed no difference between folate sufficient and deficient cells. Immunofluorescence by confocal microscopy showed that there was no significant change in the nuclear localization of NF-?B in folate deficient conditions even when stimulated with TNF-a. Analysis of reactive oxygen species (ROS) using a fluorescent assay with flow cytometry revealed that there was more fluorescence per cell in folate deficient cells at day 11. This increase in ROS could be one of the mechanisms behind the increase in NF-?B activity in folate deficient cells.

Additional Information

Publication
Thesis
Language: English
Date: 2009
Keywords
Folate, NF-kappa B
Subjects
Folic acid deficiency $x Complications.
Cellular control mechanisms.
Proteins $x Metabolism.