NEAT1 is Required for the Expression of the Liver Cancer Stem Cell Marker CD44
Abstract
:1. Introduction
2. Results
2.1. Increased Expression of NEAT1 in Spheroids
2.2. Increased Spheroid Formation and CD44 Expression by Mouse Neat1v1
2.3. Decreased Spheroid Formation and CD44 Expression by NEAT1 Knockout
2.4. Increased Expression of p21CDKN1A in NEAT1-KO Cells
2.5. NEAT1v1-Induced CD44 Expression
2.6. Rescue of CD44-KO Cells by NEAT1v1
2.7. Association of NEAT1v1 with Poor Prognosis in Patients with HCC
2.8. Association of NEAT1v1 with Poor Prognosis in Patients with HCC
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. Reverse-Transcription Quantitative PCR (RT-qPCR)
4.3. Western Blot Analysis
4.4. Stable Transfection of NEAT1-Expressing Vectors
4.5. Knockout of the NEAT1 Gene
4.6. SiRNA Transfection
4.7. Cell viability Assay
4.8. Analysis of Human Samples
4.9. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
5-FU | 5-Fluorouracil |
Blst | Blasticidin |
CD44v | CD44 variant isoform |
CD44s | CD44 standard isoform |
CSC | Cancer stem cell |
EPCAM | Epithelial adhesion molecule |
GAPDH | Glyceraldehyde-3-phosphate dehydrogenase |
HCC | Hepatocellular carcinoma |
HPRT1 | Hypoxanthine phosphoribosyltransferase 1 |
KO | Knock out |
lncRNA | Long noncoding RNA |
miR | MicroRNA |
NEAT1 | Nuclear enriched abundant transcript 1 |
PTBP3 | Polypyrimidine tract-binding proteins 3 |
RT-qPCR | Reverse-transcription quantitative PCR |
SD | Standard deviation |
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Koyama, S.; Tsuchiya, H.; Amisaki, M.; Sakaguchi, H.; Honjo, S.; Fujiwara, Y.; Shiota, G. NEAT1 is Required for the Expression of the Liver Cancer Stem Cell Marker CD44. Int. J. Mol. Sci. 2020, 21, 1927. https://doi.org/10.3390/ijms21061927
Koyama S, Tsuchiya H, Amisaki M, Sakaguchi H, Honjo S, Fujiwara Y, Shiota G. NEAT1 is Required for the Expression of the Liver Cancer Stem Cell Marker CD44. International Journal of Molecular Sciences. 2020; 21(6):1927. https://doi.org/10.3390/ijms21061927
Chicago/Turabian StyleKoyama, Shigemi, Hiroyuki Tsuchiya, Masataka Amisaki, Hiromi Sakaguchi, Soichiro Honjo, Yoshiyuki Fujiwara, and Goshi Shiota. 2020. "NEAT1 is Required for the Expression of the Liver Cancer Stem Cell Marker CD44" International Journal of Molecular Sciences 21, no. 6: 1927. https://doi.org/10.3390/ijms21061927