Integrated Transcriptome and Metabolome Analysis Reveals the Regulatory Mechanisms of FASN in Geese Granulosa Cells
Abstract
:1. Introduction
2. Results
2.1. Efficiency of FASN Overexpression and Interference
2.2. Metabolomics Analysis
2.3. Expression Profile of DEGs
2.4. Functional Enrichment Analysis of DEGs
2.5. Network Analysis for DEGs in FASN-Overexpressed phGCs and hGCs
2.6. Combined Metabolome and Transcriptome Analysis
3. Discussion
4. Materials and Methods
4.1. Experimental Animals
4.2. GC Culture and Transfection
4.3. Metabolomics Detection
4.4. Metabolites Identification
4.5. Differential Metabolite Analysis
4.6. RNA-Seq and Functional Analysis of the DEGs
4.7. qRT-PCR for Validation of the DEGs
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Compared Samples | Num. of Total Sig. | Num. of Sig. Up | Num. of Sig. Down |
---|---|---|---|
ph_OE vs. ph_OENC | 63 | 59 | 4 |
po_OE vs. po_OENC | 54 | 33 | 21 |
ph_SI vs. ph_SINC | 36 | 19 | 17 |
po_SI vs. po_SINC | 35 | 17 | 18 |
Group | Map ID | Map Title | p-Value |
---|---|---|---|
ph_OE vs. ph_OENC | map00240 | Pyrimidine metabolism | 0.019931862 |
map00520 | Amino sugar and nucleotide sugar metabolism | 0.042016807 | |
map00480 | Glutathione metabolism | 0.031746032 | |
ph_SI vs. ph_SINC | map00780 | Biotin metabolism | 0.015873016 |
po_SI vs. po_SINC | map04726 | Serotonergic synapse | 0.001424299 |
map04080 | Neuroactive ligand-receptor interaction | 0.001914757 | |
map04540 | Gap junction | 0.004199855 | |
map04742 | Taste transduction | 0.004199855 | |
map04750 | Inflammatory mediator regulation of TRP channels | 0.004199855 | |
map04024 | cAMP signaling pathway | 0.008255516 | |
map04721 | Synaptic vesicle cycle | 0.008255516 | |
map03320 | PPAR signaling pathway | 0.042016807 | |
map04261 | Adrenergic signaling in cardiomyocytes | 0.042016807 | |
map04270 | Vascular smooth muscle contraction | 0.042016807 | |
map04924 | Renin secretion | 0.042016807 | |
map04970 | Salivary secretion | 0.042016807 |
Group | Num. of DEG | Num. of Sig. Up | Num. of Sig. Down |
---|---|---|---|
ph_OE vs. ph_OENC | 1099 | 724 | 375 |
po_OE vs. po_OENC | 591 | 373 | 128 |
ph_SI vs. ph_SINC | 224 | 102 | 122 |
po_SI vs. po_SINC | 161 | 70 | 91 |
Group | Experimental Group | Control Group |
---|---|---|
ph_OE vs. ph_OENC | phGCs transfected with pEGFP-N1-FASN | phGCs transfected with pEGFP-N1 |
po_OE vs. po_OENC | hGCs transfected with pEGFP-N1-FASN | hGCs transfected with pEGFP-N1 |
ph_SI vs. ph_SINC | phGCs transfected with siRNA-FASN | phGCs transfected with siRNA-NC |
po_SI vs. po_SINC | hGCs transfected with siRNA-FASN | hGCs transfected with siRNA-NC |
Genes | AccessionNumber | Primer (5′-3′) | Tm (°C) | Size (bp) |
---|---|---|---|---|
TLR3 | KC292270.1 | F:CCCCCAGACTGAAGGAGGTAR:TGACATTGCTTTCCGACCGA | 63 | 129 |
EIF2AK2 | XM_013170695.1 | F:GACCTCACCATGTGGACCAGR:CTTCTTTTGCAGCGGCTTGT | 63 | 227 |
FADD | XM_013194898.1 | F:ACCATGGACCCGTTTCTGACR:AGGAAGTTGAAGAGCTCCGC | 63 | 149 |
IKBKE | XM_013189616.1 | F:GGAGCATCGAGTGGAGCTAC R:TCGAACCCCCAGCATTTCTC | 65 | 124 |
IRF7 | XM_013174398.1 | F:CAGCAGCGACATCGAGATCTR:TCGCTGTTCTTGGAGTGGTC | 63 | 156 |
BID | XM_013172977.1 | F:CGAGAGAAGGCCATGTTGGTR:GCTGGAGATCTCTCACTCGC | 63 | 162 |
BIRC2 | XM_013178135.1 | F:CTCTGGTACCTGGAGCTCCT R:AGAATGGACTCACTGCTGGC | 63 | 179 |
MAP3K8 | XM_013191750.1 | F:ATTCCTCGGGGAGCTTTTGGR:GATGGATCGTCTCGTCCCAC | 63 | 184 |
STAT1 | XM_013187488.1 | F:GGATCTCAAAACGAGCCCCAR:ATGTAGCCGTTTCCCTTGGG | 63 | 245 |
TNFAIP3 | XM_013175520.1 | F:GACAGCCCGAAAGAACAGGAR:CACAGCTCCTCACTTCTGCA | 63 | 107 |
AGT | XM_048045900.1 | F:AAACAGGGATGAGGAGCTGCR:TGGCCTCCACAAAGGTGTTT | 63 | 182 |
HMOX1 | XM_013181078.2 | F:GGAGGAGGCCAAGAAAGCATR:GCCTTTGACTGCACAAGGTG | 63 | 105 |
β-actin * | M26111.1 | F:CAACGAGCGGTTCAGGTGTR:TGGAGTTGAAGGTGGTCTCG | 60 | 170 |
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Chen, X.; Huang, K.; Hu, S.; Lan, G.; Gan, X.; Gao, S.; Deng, Y.; Hu, J.; Li, L.; Hu, B.; et al. Integrated Transcriptome and Metabolome Analysis Reveals the Regulatory Mechanisms of FASN in Geese Granulosa Cells. Int. J. Mol. Sci. 2022, 23, 14717. https://doi.org/10.3390/ijms232314717
Chen X, Huang K, Hu S, Lan G, Gan X, Gao S, Deng Y, Hu J, Li L, Hu B, et al. Integrated Transcriptome and Metabolome Analysis Reveals the Regulatory Mechanisms of FASN in Geese Granulosa Cells. International Journal of Molecular Sciences. 2022; 23(23):14717. https://doi.org/10.3390/ijms232314717
Chicago/Turabian StyleChen, Xi, Kailiang Huang, Shenqiang Hu, Gang Lan, Xiang Gan, Shanyan Gao, Yan Deng, Jiwei Hu, Liang Li, Bo Hu, and et al. 2022. "Integrated Transcriptome and Metabolome Analysis Reveals the Regulatory Mechanisms of FASN in Geese Granulosa Cells" International Journal of Molecular Sciences 23, no. 23: 14717. https://doi.org/10.3390/ijms232314717