NACC1, as a Target of MicroRNA-331-3p, Regulates Cell Proliferation in Urothelial Carcinoma Cells
Abstract
:1. Introduction
2. Results
2.1. NACC1 Is Overexpressed in Various Stages of UC in the Urinary Bladder
2.2. NACC1 Inhibition Suppresses Proliferation in UC Cells
2.3. NACC1 Is One of the Putative Targets of MiR-331-3p
2.4. NACC1 Inhibits UC Cell Invasion and Migration In Vitro
3. Discussion
4. Materials and Methods
4.1. Cell Lines
4.2. siRNA and miRNA Inhibitor/Precursor Transfection in UC Cells
4.3. Quantitative RT-PCR Analysis of miRNA and mRNA
- NACC1 sense 5′-CTCTCCCGGCTGAACTTATCAAC-3′,
- NACC1 antisense 5′-GTACACGTTGGTGCCTGTCAC-3′;
- Actin sense 5′-CTCTTCCAGCCTTCCTTCCT-3′,
- Actin antisense 5′-AGCACTGTGTTGGCGTACAG-3′;
- CCNA1 sense 5′-ACCCCAAGAGTGGAGTTGTG-3′,
- CCNA1 antisense 5′-GGAAGGCATTTTCTGATCCA-3′;
- CCNA2 sense 5′-TTATTGCTGGAGCTGCCTTT-3′,
- CCNA2 antisense 5′-ACTGTTGTGCATGCTGTGGT-3′;
- CCNB1 sense 5′-CGGGAAGTCACTGGAAACAT-3′,
- CCNB1 antisense 5′-AAACATGGCAGTGACACCAA-3′;
- CCNB2 sense 5′-TGGAAAAGTTGGCTCCAAAG-3′,
- CCNB2 antisense 5′-CCTCCAGCTGCCTGAGATAC-3′.
4.4. Cell Proliferation Assay
4.5. Senescence-Associated β-Galactosidase (SA-β-gal) Assay
4.6. Cell Invasion Assay and Wound Healing Assay
4.7. Tissue Samples
4.8. Immunohistochemistry
4.9. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Morita, K.; Fujii, T.; Itami, H.; Uchiyama, T.; Nakai, T.; Hatakeyama, K.; Sugimoto, A.; Miyake, M.; Nakai, Y.; Tanaka, N.; et al. NACC1, as a Target of MicroRNA-331-3p, Regulates Cell Proliferation in Urothelial Carcinoma Cells. Cancers 2018, 10, 347. https://doi.org/10.3390/cancers10100347
Morita K, Fujii T, Itami H, Uchiyama T, Nakai T, Hatakeyama K, Sugimoto A, Miyake M, Nakai Y, Tanaka N, et al. NACC1, as a Target of MicroRNA-331-3p, Regulates Cell Proliferation in Urothelial Carcinoma Cells. Cancers. 2018; 10(10):347. https://doi.org/10.3390/cancers10100347
Chicago/Turabian StyleMorita, Kohei, Tomomi Fujii, Hiroe Itami, Tomoko Uchiyama, Tokiko Nakai, Kinta Hatakeyama, Aya Sugimoto, Makito Miyake, Yasushi Nakai, Nobumichi Tanaka, and et al. 2018. "NACC1, as a Target of MicroRNA-331-3p, Regulates Cell Proliferation in Urothelial Carcinoma Cells" Cancers 10, no. 10: 347. https://doi.org/10.3390/cancers10100347