Silencing Stem Cell Factor Gene in Fibroblasts to Regulate Paracrine Factor Productions and Enhance c-Kit Expression in Melanocytes on Melanogenesis
Abstract
:1. Introduction
2. Results
2.1. The Validation of Short Interfering RNA Knockdown Efficiency
2.2. The Proliferation of Fibroblasts Was Not Suppressed by SCF Gene Silencing
2.3. The Variations in Paracrine Factor Expressions through SCF Silencing in Fibroblasts
2.4. The Variations in Paracrine Factor Expressions through SCF Silencing in Fibroblasts Using the UVB-Stimulated Model
2.5. The Influence on Melanocytes When Treated with Conditioned Medium
2.6. The Variation in Melanin Content in Melanocytes
2.7. c-Kit Activation and SCF Regulation through Autocrine Mechanism
3. Discussion and Conclusions
4. Materials and Methods
4.1. Reagents and Materials
4.2. Cell Cultures
4.3. Transfection of SCF-Targeted siRNA into Fibroblasts
4.4. MTT Assay in Cell Viability
4.5. UVB Exposure to Fibroblasts and Melanocytes Treated with Conditioned Medium
4.6. Total RNA Isolation and Extraction
4.7. Detection of mRNA Gene Expression Using qPCR
4.8. Melanin Content Assessment
4.9. Statistical Analysis
Author Contributions
Acknowledgments
Conflicts of Interest
References
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Target sequence 1: GGAAUCGUGUGACUAUAA |
Target sequence 2: AUAAGUAUGUUGCAAGAGA |
Target sequence 3: UAAGCGAGAUGGUAGUACA |
Target sequence 4: GCUUUAUAGUUGCCGAUUA |
SCF |
Forward: 5′-CTGCCAACGATCCTATCTTCCT-3′ |
Reverse: 5′-GGTTATGTCCAATGGGTGCATT-3′ |
HGF |
Forward: 5′-TCCCTACCTCTCTCGCTGTCT-3′ |
Reverse: 5′-GGGTAAGGGCCAGCATGTA-3′ |
DKK-1 |
Forward: 5′-GCGGGAATAAGTACCAGACCAT-3′ |
Reverse: 5′-TGCAGGCGAGACAGATTTG-3′ |
NRG |
Forward: 5′-AGGCCAGGACCCTATTATTTC-3′ |
Reverse: 5′-TTCAGATTGAGCCCTAGAGACA-3′ |
CRH |
Forward: 5′-AGGCACCGGAGAGAGAAAG-3′ |
Reverse: 5′-GTTTCCTGTTGCTGTGAGCTT-3′ |
ET-1 |
Forward: 5′-TGGTTCCTGACTGGCAAAG-3′ |
Reverse: 5′-GGAAGCCAGTGAAGATGGTT-3′ |
Tyrosinase |
Forward: 5′-CTGCCAACGATCCTATCTTCCT-3′ |
Reverse: 5′-GGTTATGTCCAATGGGTGCATT-3′ |
MITF |
Forward: 5′-TTGGTGCCACCTAAAACATTGT-3′ |
Reverse: 5′-CCGTTGGGCTTGCTGTATG-3′ |
ERK-1 |
Forward: 5′-CAACACCACCTGCGACCTT-3′ |
Reverse: 5′-GCCACATACTCCGTCAGGAA-3′ |
ERK-2 |
Forward: 5′-CGGTGTTCTTCTTCCCAGTTC-3′ |
Reverse: 5′-AAAGCCACAACTACCAGAAACC-3′ |
Pmel17 |
Forward: 5′-GGATGGTACAGCCACCTTAAGG-3′ |
Reverse: 5′-CAGGATCTCGGCACTTTCAATAC-3′ |
Myo5a |
Forward: 5′-GCCCAGATTGTGAAAGTGTTGA-3′ |
Reverse: 5′-CCTGTCTCGTAAACGCATCTGT-3′ |
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Li, P.-H.; Liu, L.-H.; Chang, C.-C.; Gao, R.; Leung, C.-H.; Ma, D.-L.; David Wang, H.-M. Silencing Stem Cell Factor Gene in Fibroblasts to Regulate Paracrine Factor Productions and Enhance c-Kit Expression in Melanocytes on Melanogenesis. Int. J. Mol. Sci. 2018, 19, 1475. https://doi.org/10.3390/ijms19051475
Li P-H, Liu L-H, Chang C-C, Gao R, Leung C-H, Ma D-L, David Wang H-M. Silencing Stem Cell Factor Gene in Fibroblasts to Regulate Paracrine Factor Productions and Enhance c-Kit Expression in Melanocytes on Melanogenesis. International Journal of Molecular Sciences. 2018; 19(5):1475. https://doi.org/10.3390/ijms19051475
Chicago/Turabian StyleLi, Pin-Hui, Li-Heng Liu, Cheng-Chung Chang, Rong Gao, Chung-Hang Leung, Dik-Lung Ma, and Hui-Min David Wang. 2018. "Silencing Stem Cell Factor Gene in Fibroblasts to Regulate Paracrine Factor Productions and Enhance c-Kit Expression in Melanocytes on Melanogenesis" International Journal of Molecular Sciences 19, no. 5: 1475. https://doi.org/10.3390/ijms19051475