Upregulated microRNA-106a Promotes Porcine Preadipocyte Proliferation and Differentiation by Targeting Different Genes
Abstract
:1. Introduction
2. Materials and Methods
2.1. Isolation and Culture of Porcine Preadipocytes
2.2. Transfection of miRNA Agomir
- Ssc-NC sense: 5-UUCUCCGAACGUGUCACGUTT-3;
- Ssc-antisense: 5-ACGUGACACGUUCGGAGAATT-3;
- Ssc-miR-106a sense: 5-AAAAGUGCUUACAGUGCAGGUAGC-3;
- Ssc-antisense: 5-UACCUGCACUGUAAGCACUUUUUU-3.
2.3. RNA Extractions and RT-qPCR
2.4. Western Blot Analysis
2.5. 5-Ethynyl-20-Deoxyuridine (EdU) Staining
2.6. Cell Counting Kit-8 (CCK-8)
2.7. Flow Cytometry
2.8. Luciferase Reporter Assay
2.9. BODIPY Staining of Lipid Droplets
2.10. Oil Red O Staining and Dye Extraction Analysis
2.11. Bioinformatics Method
2.12. Statistical Analysis
3. Results
3.1. miR-106a is Widely Expressed in Various Porcine Tissues and is Closely Related to The Cell Cycle and Adipose Metabolism
3.2. Upregulated miR-106a Promotes the Proliferation of Porcine Preadipocytes
3.3. miR-106a Can Target p21 Instead of BAMBI in Proliferative Phase
3.4. miR-106a Promotes the Differentiation of Porcine Preadipocytes
3.5. miR-106a Can Target BAMBI Instead of p21 during Porcine Preadipocytes Differentiation.
4. Discussion
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Gene | Primer Sequence | Length | Temp (°C) |
---|---|---|---|
PPARγ | F: AGGACTACCAAAGTGCCATCAAA R: GAGGCTTTATCCCCACAGACAC | 164 | 60 |
aP2 | F: GAGCACCATAACCTTAGATGGA R: AAATTCTGGTAGCCGTGACA | 142 | 60 |
C/EBPβ | F: GCACAGCGACGAGTACAAGA R: TATGCTGCGTCTCCAGGTTG | 98 | 60 |
p21 | F:ACGTCTCAGGAGGACCATGT R:AGAAGATCAGCCGGCGTTTG | 166 | 60 |
BAMBI | F:AGGACAAGGCAACAGGTATTAGC R: GAACCACAACTCTTTGGAGGAAG | 96 | 60 |
β-actin | F: GGACTTCGAGCAGGAGATGG R: AGGAAGGAGGGCTGGAAGAG | 138 | 60 |
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Huang, K.; Shi, X.; Wang, J.; Yao, Y.; Peng, Y.; Chen, X.; Li, X.; Yang, G. Upregulated microRNA-106a Promotes Porcine Preadipocyte Proliferation and Differentiation by Targeting Different Genes. Genes 2019, 10, 805. https://doi.org/10.3390/genes10100805
Huang K, Shi X, Wang J, Yao Y, Peng Y, Chen X, Li X, Yang G. Upregulated microRNA-106a Promotes Porcine Preadipocyte Proliferation and Differentiation by Targeting Different Genes. Genes. 2019; 10(10):805. https://doi.org/10.3390/genes10100805
Chicago/Turabian StyleHuang, Kuilong, Xin’e Shi, Jie Wang, Ying Yao, Ying Peng, Xiaochang Chen, Xiao Li, and Gongshe Yang. 2019. "Upregulated microRNA-106a Promotes Porcine Preadipocyte Proliferation and Differentiation by Targeting Different Genes" Genes 10, no. 10: 805. https://doi.org/10.3390/genes10100805