Downregulation of miR-140-3p Contributes to Upregulation of CD38 Protein in Bronchial Smooth Muscle Cells
Abstract
:1. Introduction
2. Results
2.1. Differentially Expressed miRNAs in BSMs of the Antigen-Induced AHR Mice
2.2. Change in miR-140-3p Level in BSMs of the Antigen-Induced AHR Mice
2.3. CD38 mRNA as a Predicted Target of miR-140-3p
2.4. Upregulation of CD38 Protein in BSMs of the Antigen-Induced AHR Mice
2.5. Effect of Inhibition of miR-140-3p on CD38 Protein Expression in Cultured Human BSM Cells (hBSMCs)
2.6. Effect of Inhibition of miR-140-3p on CD38 Protein Exp 2.6. Effects of IL-13 on Expression Levels of CD38 and miR-140-3p in Cultured Human BSM Cells (hBSMCs)
2.7. Effects of Cyclic ADP Ribose (cADPR) and Its Antagonist 8-Bromo-cADPR (8-Br-cADPR) on BSM Function
3. Discussion
4. Materials and Methods
4.1. Mouse Model of Allergic Bronchial Asthma
4.2. miRNA Microarray Analyses
4.3. Cell Culture and Sample Collection
4.4. Transfection of miR-140-3p Inhibitor and Mimic
4.5. Quantitative RT-PCR Analyses
4.6. Western Blot Analyses
4.7. Data and Statistical Analyses
Author Contributions
Funding
Conflicts of Interest
Abbreviations
ACh | acetylcholine |
AHR | airway hyperresponsiveness |
ANOVA | analysis of variance |
ASM | airway smooth muscle |
BSM | bronchial smooth muscle |
cADPR | cyclic ADP-ribose |
COPD | chronic obstructive pulmonary disease |
CT | threshold cycle |
GAPDH | glyceraldehyde-3-phosphate dehydrogenase |
hBSMC | human bronchial smooth muscle cell |
hEGF | human epidermal growth factor |
hFGF-b | human fibroblast growth factor-basic |
IL-13 | interleukin-13 |
LNA | locked nucleic acid |
miRNA | microRNA |
mRNA | messenger RNA |
NAD | nicotinamide adenine dinucleotide |
OA | ovalbumin |
pre-miRNA | precursor miRNA |
PCR | polymerase chain reaction |
pri-miRNA | primary miRNA |
PVDF | polyvinylidene difluoride |
RT | reverse transcription |
RT-qPCR | quantitative real-time RT-PCR |
snRNA | small nuclear RNA |
UTR | untranslated region |
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miRNA (mmu) | Accession | Mature Sequence | Human Expression | AHR/Cont |
---|---|---|---|---|
miR-140-3p | MIMAT0000152 | 5′-UACCACAGGGUAGAACCACGG-3′ | Yes | 0.448 |
miR-140-5p | MIMAT0000151 | 5′-CAGUGGUUUUACCCUAUGGUAG-3′ | Yes | 0.449 |
miR-133a-3p | MIMAT0000145 | 5′-UUUGGUCCCCUUCAACCAGCUG-3′ | Yes | 0.465 |
miR-1971 | MIMAT0009446 | 5′-GUAAAGGCUGGGCUGAGA-3′ | No | 2.004 |
miR-142-3p | MIMAT0000155 | 5′-UGUAGUGUUUCCUACUUUAUGGA-3′ | Yes | 2.101 |
miR-669c-5p | MIMAT0003479 | 5′-AUAGUUGUGUGUGGAUGUGUGU-3′ | No | 2.123 |
miR-1897-5p | MIMAT0007864 | 5′-CUUUGGAUGGAGAAAGAGGGGG-3′ | No | 2.257 |
miR-300-5p | MIMAT0004578 | 5′-UUGAAGAGAGGUUAUCCUUUGU-3′ | Yes | 2.262 |
miR-1196-5p | MIMAT0005857 | 5′-AAAUCUACCUGCCUCUGCCU-3′ | No | 2.488 |
miR-302a-3p | MIMAT0000380 | 5′-UAAGUGCUUCCAUGUUUUGGUGA-3′ | Yes | 2.525 |
miR-133b-3p | MIMAT0000769 | 5′-UUUGGUCCCCUUCAACCAGCUA-3′ | Yes | 3.134 |
miR-1947-3p | MIMAT0017343 | 5′-GCACUGAGCUAGCUCUCCCUCC-3′ | No | 6.173 |
miR-3100-3p | MIMAT0014920 | 5′-CUGUGACACACCCGCUCCCAG-3′ | No | 6.410 |
miR-3474 | MIMAT0015646 | 5′-CCCUGGGAGGAGACGUGGAUUC-3′ | No | 12.195 |
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Chiba, Y.; Matsumoto, M.; Hanazaki, M.; Sakai, H. Downregulation of miR-140-3p Contributes to Upregulation of CD38 Protein in Bronchial Smooth Muscle Cells. Int. J. Mol. Sci. 2020, 21, 7982. https://doi.org/10.3390/ijms21217982
Chiba Y, Matsumoto M, Hanazaki M, Sakai H. Downregulation of miR-140-3p Contributes to Upregulation of CD38 Protein in Bronchial Smooth Muscle Cells. International Journal of Molecular Sciences. 2020; 21(21):7982. https://doi.org/10.3390/ijms21217982
Chicago/Turabian StyleChiba, Yoshihiko, Mayumi Matsumoto, Motohiko Hanazaki, and Hiroyasu Sakai. 2020. "Downregulation of miR-140-3p Contributes to Upregulation of CD38 Protein in Bronchial Smooth Muscle Cells" International Journal of Molecular Sciences 21, no. 21: 7982. https://doi.org/10.3390/ijms21217982