Analysis of MicroRNA Expression in Newborns with Differential Birth Weight Using Newborn Screening Cards
Abstract
:1. Introduction
2. Results
2.1. Isolation of miRNAs from Dried Blood Spots on Newborn Screening Cards (NSC)
2.2. miR-33b, miR-375 and miR-454-3p Relative Expression in Neonates with Differential Birth Weight
2.3. Prediction of Target Genes and Signaling Pathways for miR-33b, miR-375 and miR-454-3p
3. Discussion
4. Materials and Methods
4.1. Ethics Statement
4.2. RNA Purification
4.3. cDNA Synthesis
4.4. Real Time Quantitative PCR
4.5. Metabolic Pathway Analysis
4.6. Statistical Analysis
5. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
DBS | dried blood spots |
NSC | newborn screening cards |
LBW | low birth weight |
NBW | normal birth weight |
T2D | type 2 diabetes mellitus |
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KEGG Pathway | p Value | Faslse Discovery Rate Adjustment | KEGG ID |
---|---|---|---|
Axon guidance | 0.0001975 | 0.00237 | ko04360 |
cGMP-PKG signaling pathway | 0.01559 | 0.00237 | ko04022 |
Type 2 diabetes mellitus | 0.01987 | 0.00237 | ko04930 |
Adherens junction | 0.02716 | 0.00237 | ko04520 |
GnRH signaling pathway | 0.02716 | 0.04484 | ko04912 |
Glutamatergic synapse | 0.02894 | 0.04484 | ko04724 |
Pathogenic Escherichia coli infection | 0.02926 | 0.04484 | ko05130 |
Cholinergic synapse | 0.03329 | 0.04484 | ko04725 |
Amphetamine addiction | 0.03363 | 0.04484 | ko05031 |
Insulin secretion | 0.03774 | 0.04484 | ko04911 |
Nicotine addiction | 0.0423 | 0.04501 | ko05033 |
Vascular smooth muscle contraction | 0.04501 | 0.04501 | ko04270 |
KEGG Pathway | p Value | FDR Adjustment | KEGG ID |
---|---|---|---|
Glutamatergic synapse | 0.0108 | 0.03308333 | ko04724 |
Transcriptional misregulation in cancer | 0.01638 | 0.03308333 | ko05202 |
Maturity onset diabetes of the young | 0.01985 | 0.03308333 | ko04950 |
Mitogen-activated protein kinase (MAPK) signaling pathway | 0.03948 | 0.03979 | ko04010 |
Axon guidance | 0.03979 | 0.03979 | ko04360 |
KEGG Pathway | p Value | FDR Adjustment | KEGG ID |
---|---|---|---|
Endocytosis | 0.001724 | 0.012068 | ko04144 |
TGF-β signaling pathway | 0.01607 | 0.04421 | ko04350 |
Axon guidance | 0.02799 | 0.04421 | ko04360 |
FoxO signaling pathway | 0.03921 | 0.04421 | ko04068 |
p53 signaling pathway | 0.03971 | 0.04421 | ko04115 |
Proteoglycans in cancer | 0.04393 | 0.04421 | ko05205 |
Hippo signaling pathway | 0.04421 | 0.04421 | ko04390 |
Name | Sequence (5′–3′) | Accession Number of Mature miRNA |
---|---|---|
hsa-miR-33b RT stem-loop | GTTGGCTCTGGTGCAGGGTCCGAGGTATTCGCACCAGAGCCAACGCAATG | MIMAT0003301 |
hsa-miR-33b specific forward | GTTTGGGTGCATTGCTGTTG | |
hsa-miR-375 RT stem-loop | GTTGGCTCTGGTGCAGGGTCCGAGGTATTCGCACCAGAGCCAACTCACGC | MIMAT0000728 |
hsa-miR-375 specific forward | TGGTTTTTGTTCGTTCGGCT | |
hsa-miR-454-3p RT stem-loop | GTTGGCTCTGGTGCAGGGTCCGAGGTATTCGCACCAGAGCCAACACCCTA | MIMAT0003885 |
hsa-miR-454-3p specific forward | GGTGTGGTAGTGCAATATTGCTTA | |
hsa-miR-106a RT stem-loop | GTTGGCTCTGGTGCAGGGTCCGAGGTATTCGCACCAGAGCCAACCTACCT | MIMAT0000103 |
hsa-miR-106a specific forward | TGGGTAAAAGTCCTTACAGTGC | |
hsa-miR-16-5p RT stem-loop | GTTGGCTCTGGTGCAGGGTCCGAGGTATTCGCACCAGAGCCAACCGCCAA | MIMAT0000069 |
hsa-miR-16-5p specific forward | TGTTTTTTTTTGTAGCAGCACGTAAATA | |
Universal reverse primer | GTGCAGGGTCCGAGGT | NA |
Universal ProbeLibrary probe #21 | TGGCTCTG | NA |
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Rodil-Garcia, P.; Arellanes-Licea, E.D.C.; Montoya-Contreras, A.; Salazar-Olivo, L.A. Analysis of MicroRNA Expression in Newborns with Differential Birth Weight Using Newborn Screening Cards. Int. J. Mol. Sci. 2017, 18, 2552. https://doi.org/10.3390/ijms18122552
Rodil-Garcia P, Arellanes-Licea EDC, Montoya-Contreras A, Salazar-Olivo LA. Analysis of MicroRNA Expression in Newborns with Differential Birth Weight Using Newborn Screening Cards. International Journal of Molecular Sciences. 2017; 18(12):2552. https://doi.org/10.3390/ijms18122552
Chicago/Turabian StyleRodil-Garcia, Patricia, Elvira Del Carmen Arellanes-Licea, Angélica Montoya-Contreras, and Luis A. Salazar-Olivo. 2017. "Analysis of MicroRNA Expression in Newborns with Differential Birth Weight Using Newborn Screening Cards" International Journal of Molecular Sciences 18, no. 12: 2552. https://doi.org/10.3390/ijms18122552