miR-3189-3p Mimics Enhance the Effects of S100A4 siRNA on the Inhibition of Proliferation and Migration of Gastric Cancer Cells by Targeting CFL2
Abstract
:1. Introduction
2. Results
2.1. S100A4 Knockdown Leads to Decreased Expression of miR-3189-3p in MGC803 Cells
2.2. miR-3189-3p Inhibits the Proliferation of MGC803 Cells
2.3. miR-3189-3p Retards MGC803 Cells Migration
2.4. miR-3189-3p Mimics Enhanced the Effects of S100A4 siRNA on the Inhibition of Proliferation and Migration of MGC803 Cells
2.5. CFL2 Is a Direct Target Gene of miR-3189-3p in MGC803 Cells
2.6. CFL2 siRNA Inhibits the Proliferation and Migration of MGC803 Cells
2.7. CFL2 Mediates the Functional Effects of miR-3189-3p on MGC803 Cells.
2.8. CFL2 Is an Unfavorable Prognostic Factor for Gastric Cancer
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. Cell Transfection
4.3. Quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR)
4.4. Western Blotting Analysis
4.5. Dual Luciferase Reporter Assay
4.6. Cell Proliferation
4.7. Transwell Assay
4.8. Wound Healing Assay
4.9. Kaplan–Meier Plotter
4.10. Statistical Analysis
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
GC | gastric cancer |
miRNA | microRNA |
3′-UTR | 3′-untranslated region |
CFL2 | cofilin 2 |
OS | overall survival |
FP | first progression |
HR | hazard ratio |
95%CI | 95% confidence intervals |
IBMS | Institute of Basic Medical Sciences |
CAMS | Chinese Academy of Medical Sciences |
PUMC | Peking Union Medical College |
SPSS | Statistical Package for the Social Sciences |
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Name of Short Nucleotides | Sequences |
---|---|
S100A4-siRNA | 5′-GCAUCGCCAUGAUGUGUAATT-3′ |
5′-UUACACAUCAUGGCGAUGCTT-3′ | |
CFL2-siRNA | 5′-GCAAGUAAAUGGCUUGGAUTT-3′ |
5′-AUCCAAGCCAUUUACUUGCTT-3′ | |
hsa-miR-3189-3p mimics | 5′-CCCUUGGGUCUGAUGGGGUAG-3′ |
5′-ACCCCAUCAGACCCAAGGGUU-3′ | |
Negative Control (NC) | 5′-UUCUCCGAACGUGUCACGUTT-3′ |
5′-ACGUGACACGUUCGGAGAATT-3′ | |
hsa-miR-3189-3p inhibitor | 5′-CUACCCCAUCAGACCCAAGGG-3′ |
hsa-miR-3189-3p inhibitor NC | 5’-CAGUACUUUUGUGUAGUACAA-3’ |
Gene | Primer Sequence (5′–3′) |
---|---|
S100A4 | F: CCCTGGATGTGATGGTGT |
R: GTTGTCCCTGTTGCTGTC | |
GDF15 | F: CTCCAGATTCCGAGAGTTGC |
R: AGAGATACGCAGGTGCAGGT | |
pri-miR-3189 | F: CAAGCAGCCCCCATATCTAA |
R: CCAAGGGGATCCAGGATATT | |
miR-3189-3p | F: ATGCTGCCCTTGGGTCTG |
R: CACTTCCTCAGCACTTGTTGGTAT | |
GAPDH | F: ATCATCAGCAATGCCTCC |
R: CATCACGCCACAGTTTCC | |
U6 | F: ATTGGAACGATACAGAGAAGATT |
R: GGAACGCTTCACGAATTTG |
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Bian, Y.; Guo, J.; Qiao, L.; Sun, X. miR-3189-3p Mimics Enhance the Effects of S100A4 siRNA on the Inhibition of Proliferation and Migration of Gastric Cancer Cells by Targeting CFL2. Int. J. Mol. Sci. 2018, 19, 236. https://doi.org/10.3390/ijms19010236
Bian Y, Guo J, Qiao L, Sun X. miR-3189-3p Mimics Enhance the Effects of S100A4 siRNA on the Inhibition of Proliferation and Migration of Gastric Cancer Cells by Targeting CFL2. International Journal of Molecular Sciences. 2018; 19(1):236. https://doi.org/10.3390/ijms19010236
Chicago/Turabian StyleBian, Yue, Junfu Guo, Linlin Qiao, and Xiuju Sun. 2018. "miR-3189-3p Mimics Enhance the Effects of S100A4 siRNA on the Inhibition of Proliferation and Migration of Gastric Cancer Cells by Targeting CFL2" International Journal of Molecular Sciences 19, no. 1: 236. https://doi.org/10.3390/ijms19010236