Synthetic miR-143 Inhibits Growth of HER2-Positive Gastric Cancer Cells by Suppressing KRAS Networks Including DDX6 RNA Helicase
Abstract
:1. Introduction
2. Results
2.1. Expression Levels of miR-143 Were Down-Regulated in Gastric Cancer Cell Lines
2.2. Expression Levels of KRAS and Downstream Molecules Were Up-Regulated in HER2-Positive Gastric Cancer Cell Lines
2.3. Ectopic Expression of miR-143 Inhibited the Growth of MKN-7 and KATO-III Cells by Targeting KRAS and Its Related Signaling Molecules
2.4. Transfection of syn-miR-143 Induced Apoptosis in MKN-7 Cells and Cell-Cycle Arrest in KATO-III Cells
2.5. KRAS and Its Related Molecules are Targets of syn-miR-143
2.6. DDX6 Is a Direct Target Gene of syn-miR-143 and syn-miR-143 Down-Regulated the Expression Level of HER2 through Targeting DDX6
2.7. Knockdown of KRAS Alone Was Not Sufficient to Suppress the Cell Growth and the Effector Signals of KRAS in HER2-Positive Gastric Cancer Cell Lines
2.8. Syn-miR-143 Exhibited an Anti-Tumor Effect on Xenografted Tumors in Nude Mice
3. Discussion
4. Material and Methods
4.1. Cell Culture and Cell Viability
4.2. Cell Transfection Experiment with miRNA or siRNA
4.3. Western Blotting Analysis
4.4. Quantitative RT-PCR
4.5. Dual luciferase Reporter Assay
4.6. Hoechst 33352 Staining
4.7. Cell-Cycle Analysis
4.8. Human Tumor Xenograft Model
4.9. Statistics
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
DDX6 | DEAD/H-box RNA helicase 6 |
Syn-miR-143 | synthetic miR-143 |
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Tokumaru, Y.; Tajirika, T.; Sugito, N.; Kuranaga, Y.; Shinohara, H.; Tsujino, T.; Matsuhashi, N.; Futamura, M.; Akao, Y.; Yoshida, K. Synthetic miR-143 Inhibits Growth of HER2-Positive Gastric Cancer Cells by Suppressing KRAS Networks Including DDX6 RNA Helicase. Int. J. Mol. Sci. 2019, 20, 1697. https://doi.org/10.3390/ijms20071697
Tokumaru Y, Tajirika T, Sugito N, Kuranaga Y, Shinohara H, Tsujino T, Matsuhashi N, Futamura M, Akao Y, Yoshida K. Synthetic miR-143 Inhibits Growth of HER2-Positive Gastric Cancer Cells by Suppressing KRAS Networks Including DDX6 RNA Helicase. International Journal of Molecular Sciences. 2019; 20(7):1697. https://doi.org/10.3390/ijms20071697
Chicago/Turabian StyleTokumaru, Yoshihisa, Toshihiro Tajirika, Nobuhiko Sugito, Yuki Kuranaga, Haruka Shinohara, Takuya Tsujino, Nobuhisa Matsuhashi, Manabu Futamura, Yukihiro Akao, and Kazuhiro Yoshida. 2019. "Synthetic miR-143 Inhibits Growth of HER2-Positive Gastric Cancer Cells by Suppressing KRAS Networks Including DDX6 RNA Helicase" International Journal of Molecular Sciences 20, no. 7: 1697. https://doi.org/10.3390/ijms20071697