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Characterization of the LYCD-Dependent Transcriptional Response in the THP-1 Cell Culture Monocytes

Osterburg, Andrew Robert
http://rave.ohiolink.edu/etdc/view?acc_num=ucin1123870203

Abstract Details

2005, PhD, University of Cincinnati, Arts and Sciences : Biological Sciences.
Live Yeast Cell Derivative (LYCD) is a medicinal extract of Saccharomyces cerevisiae that has demonstrated efficacy in improving the rate and quality of wound healing in mouse and human systems. However, the mechanisms by which LYCD promotes healing are largely uncharacterized. We have shown that LYCD has effects on the transcriptional profile of the human monocytic cell line THP-1. In cell culture, LYCD treatments induced a dose-dependent increase in the relative expression of the proto-oncogene c-Fos (6 to 44-fold) in complete (10% FBS) and low serum (0.1% FBS) media after 30 minutes of exposure. Furthermore, protein levels of c-Fos rise at 30 minutes of LYCD exposure and remain detectable through at least 120 minutes of LYCD exposure. However, the relative abundance of the c-Fos transcript returns to basal levels by 120 min. LYCD also induced expression of c-Jun with maximal expression of 3- fold at 60 minutes of exposure. Pretreatments with epidermal growth factor receptor (EGFR) kinase inhibitor AG-1478 and the MEK1 inhibitor PD98059 blocked the LYCD-dependent increases in c-Fos expression. Consistent with signaling through the EGFR we have demonstrated, by RT-PCR, the presence of the mRNA for the EGFR (ErbB1/HER1) in THP-1 cells. Additionally, with microarrays representing 22,102-70mer human oligonucleotides we measured the transcriptional profile of these cells after 4 hours of LYCD treatment. There were 116 genes with a statistically significant response to LYCD treatments of which 67 were up-regulated by at least 1.2-fold and 49 were down-regulated by at least 1.2-fold. Six of these genes (JAK2, CYP1B1, c-Fos, HMOX1, VEGF-C, TNF-alpha) were confirmed by real-time quantitative PCR. The microarray gene list was screened for significant enrichments of Gene Ontology categories. From this evaluation we found that LYCD appears to modulate; cell cycle progression, cell death, oxidoreductase metabolism, response to stress, growth factor signaling and vasculature development. Our data suggests that LYCD may modulate a number of monocyte functions and that this may help to promote wound healing.
Dr. Stephen Keller (Advisor)
156 p.
Biology, Molecular
Live Yeast Cell Derivative; monocyte; medicinal; microarray; real-time PCR; wound healing

Recommended Citations

Citations

  • Osterburg, A. R. (2005). Characterization of the LYCD-Dependent Transcriptional Response in the THP-1 Cell Culture Monocytes [Doctoral dissertation, University of Cincinnati]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1123870203

    APA Style (7th edition)

  • Osterburg, Andrew. Characterization of the LYCD-Dependent Transcriptional Response in the THP-1 Cell Culture Monocytes. 2005. University of Cincinnati, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ucin1123870203.

    MLA Style (8th edition)

  • Osterburg, Andrew. "Characterization of the LYCD-Dependent Transcriptional Response in the THP-1 Cell Culture Monocytes." Doctoral dissertation, University of Cincinnati, 2005. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1123870203

    Chicago Manual of Style (17th edition)

ucin1123870203
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© 2005, all rights reserved.
This open access ETD is published by University of Cincinnati and OhioLINK.