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Induction of MiR-17-3p and MiR-106a [corrected] by TNFα and LPS
1Signosis, Inc., 528 Weddell Drive #5, Sunnyvale, CA 94089, USA. xinjiang@signosisinc.com
Abstract:
MicroRNAs (miRNAs), small non-coding molecules, regulate gene expression in response to stimuli. Lipopolysaccharide (LPS) was reported to induce the expression of miR-146 and miR-155 in HL-60. The effects of LPS and the related stimulus, tumour necrosis factor alpha (TNFα), on miRNA expression required to be further studied. Using T7-oligo ligation assay (OLA)-based miRNA array, we profiled the expression of 132 miRNAs and identified a number of TNFα-regulated miRNAs in HeLa cells, including miR-17-3p and miR-106a. TNFα induction of miR-17-3p and miR-106a was verified by Northern blot analysis with RNU48 normalization. Northern blot analysis also showed that LPS was able to induce the expression of both miR-17-3p and miR-106a in HeLa cells. Furthermore, both array assay and Northern blot analysis showed that the expression levels of miR-146 and miR-155 were either low or undetectable in HeLa cells and TNFα- and LPS-mediated induction of these two miRNAs was not found. Luciferase reporter analysis confirmed the induction of miR-17-3p and miR-106a in response to TNFα and LPS treatment in HeLa cells. These results suggested that the expression of miR-17-3p and miR-106a is regulated by TNFα and LPS in HeLa cells.
Insights
Tumor necrosis factor alpha (TNFα) and lipopolysaccharide (LPS) regulate specific microRNAs (miRNAs). This study identifies TNFα and LPS induction of miR-17-3p and miR-106a in HeLa cells.
Area of Science:
- Molecular Biology
- Genetics
- Immunology
Background:
- MicroRNAs (miRNAs) are small non-coding RNA molecules that regulate gene expression.
- Lipopolysaccharide (LPS) and tumor necrosis factor alpha (TNFα) are known immune stimuli.
- Previous studies indicated LPS induces miR-146 and miR-155 in HL-60 cells, but the effects of LPS and TNFα on miRNA expression in other cell types needed further investigation.
Purpose of the Study:
- To investigate the effects of TNFα and LPS on miRNA expression profiles in HeLa cells.
- To identify specific miRNAs regulated by TNFα and LPS in HeLa cells.
- To validate the expression changes of identified miRNAs using independent methods.
Main Methods:
- miRNA expression profiling using T7-oligo ligation assay (OLA)-based miRNA array.
- Verification of miRNA expression using Northern blot analysis with RNU48 normalization.
- Luciferase reporter assays to confirm miRNA regulation.
Main Results:
- The miRNA array identified several TNFα-regulated miRNAs in HeLa cells, including miR-17-3p and miR-106a.
- Northern blot analysis confirmed TNFα-induced expression of miR-17-3p and miR-106a.
- LPS also induced the expression of miR-17-3p and miR-106a in HeLa cells, as confirmed by both array and Northern blot.
- miR-146 and miR-155 were found at low or undetectable levels in HeLa cells, with no induction observed upon TNFα or LPS treatment.
- Luciferase reporter assays validated the induction of miR-17-3p and miR-106a by TNFα and LPS.
Conclusions:
- TNFα and LPS regulate the expression of miR-17-3p and miR-106a in HeLa cells.
- The expression of miR-146 and miR-155 is not significantly affected by TNFα or LPS in HeLa cells.
- These findings highlight cell-type-specific responses of miRNA expression to immune stimuli.
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