TY - JOUR
T1 - Doxycycline Changes the Transcriptome Profile of mIMCD3 Renal Epithelial Cells
AU - Jung, Hyun Jun
AU - Coleman, Richard
AU - Woodward, Owen M.
AU - Welling, Paul A.
N1 - Funding Information:
This work was supported by grants from the National Institute of Diabetes and Digestive and Kidney Diseases (Grants DK054231, 1U54DK126114, and DK093501) and the Leducq Foundation.
Funding Information:
Next-generation DNA sequencing for RNA-Seq transcriptomic analysis was performed by the Genomic Resource Center, Institute for Genome Sciences, University of Maryland School of Medicine (Director: Dr. Lisa DeShong Sadzewicz).
Publisher Copyright:
© Copyright © 2021 Jung, Coleman, Woodward and Welling.
PY - 2021/11/5
Y1 - 2021/11/5
N2 - Tetracycline-inducible gene expression systems have been used successfully to study gene function in vivo and in vitro renal epithelial models but the effects of the common inducing agent, doxycycline (DOX), on gene expression are not well appreciated. Here, we evaluated the DOX effects on the transcriptome of a widely used renal epithelial cell model, mIMCD3 cells, to establish a reference. Cells were grown on permeable filter supports in the absence and presence of DOX (3 or 6 days), and genome-wide transcriptome profiles were assessed using RNA-Seq. We found DOX significantly altered the transcriptome profile, changing the abundance of 1,549 transcripts at 3 days and 2,643 transcripts at 6 days. Within 3 days of treatment, DOX significantly decreased the expression of multiple signaling pathways (ERK, cAMP, and Notch) that are associated with cell proliferation and differentiation. Genes associated with cell cycle progression were subsequently downregulated in cells treated with DOX for 6 days, as were genes involved in cellular immune response processes and several cytokines and chemokines, correlating with a remarkable repression of genes encoding cell proliferation markers. The results provide new insight into responses of renal epithelial cells to DOX and a establish a resource for DOX-mediated gene expression systems.
AB - Tetracycline-inducible gene expression systems have been used successfully to study gene function in vivo and in vitro renal epithelial models but the effects of the common inducing agent, doxycycline (DOX), on gene expression are not well appreciated. Here, we evaluated the DOX effects on the transcriptome of a widely used renal epithelial cell model, mIMCD3 cells, to establish a reference. Cells were grown on permeable filter supports in the absence and presence of DOX (3 or 6 days), and genome-wide transcriptome profiles were assessed using RNA-Seq. We found DOX significantly altered the transcriptome profile, changing the abundance of 1,549 transcripts at 3 days and 2,643 transcripts at 6 days. Within 3 days of treatment, DOX significantly decreased the expression of multiple signaling pathways (ERK, cAMP, and Notch) that are associated with cell proliferation and differentiation. Genes associated with cell cycle progression were subsequently downregulated in cells treated with DOX for 6 days, as were genes involved in cellular immune response processes and several cytokines and chemokines, correlating with a remarkable repression of genes encoding cell proliferation markers. The results provide new insight into responses of renal epithelial cells to DOX and a establish a resource for DOX-mediated gene expression systems.
KW - RNA-seq
KW - cell proliferation
KW - doxycycline
KW - mIMCD3
KW - transcriptional response
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U2 - 10.3389/fphys.2021.771691
DO - 10.3389/fphys.2021.771691
M3 - Article
C2 - 34803745
AN - SCOPUS:85119410868
SN - 1664-042X
VL - 12
JO - Frontiers in Physiology
JF - Frontiers in Physiology
M1 - 771691
ER -