Targeting Nuclear Exporter Protein XPO1/CRM1 in Gastric Cancer
Abstract
:1. Introduction
2. Results
2.1. XPO1 Is Elevated in Gastric Adenocarcinoma Compared to Normal Tissues
2.2. Selective Inhibitors of Nuclear Export (SINE) Target XPO1 and Inhibit Cell Viability
2.3. SINE Interfere with Processes Necessary for Gastric Cancer Cell Survival
2.4. SINE Induce Apoptosis in Gastric Cancer Cell Lines
2.5. XPO1 Inhibition Causes Perturbation of Small RNA Expression
2.6. KPT-330 Exhibits Synergy with Paclitaxel In Vitro and In Vivo
3. Discussion
4. Materials and Methods
4.1. Cell Lines, Culture Conditions and Reagents
4.2. Annexin V and 7-Aminoactinomycin D (7AAD) Staining Assay
4.3. Cell Cycle Arrest
4.4. Colony Formation
4.5. Growth Inhibition by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide Assay (MTT)
4.6. Growth Inhibition using Trypan Blue Cell Viability Assay
4.7. Immunofluorescence Assay
4.8. In Vivo Model
4.9. miRNA Transfection
4.10. RNA Extraction
4.11. Non-Coding RNA Data Analysis (miRNA)
4.12. Non-Coding Data Analysis (piRNA)
4.13. siRNA Interference
4.14. The Human Protein Atlas Database Search
4.15. Western Blotting
4.16. 3D Spheroid Culture
4.17. ImageJ Analysis
4.18. Statistical Analysis
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
XPO1 | Exportin 1 |
miRNA | microRNAs |
piRNA | piwiRNA |
SINE | Selective inhibitors of nuclear export |
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microRNA | Expression | Log2 Fold Change |
---|---|---|
Hsa-miR-130a-3p | Downregulated | −2.16 |
Hsa-miR-7974 | Downregulated | −1.97 |
Hsa-let-7c-3p | Downregulated | −1.95 |
Hsa-miR-33b-3p_R-1 | Downregulated | −1.89 |
Hsa-miR-12135_1ss9TC | Downregulated | −1.80 |
Hsa-miR-409-5p | Upregulated | 2.05 |
Hsa-miR-129-1-3p | Upregulated | 2.09 |
Hsa-miR-548az-5p_L-1R+1 | Upregulated | 2.18 |
Hsa-miR-147b-5p | Upregulated | 2.24 |
Hsa-miR-376c-3p | Upregulated | 2.96 |
piRNA | Expression | Log2 Fold Change |
---|---|---|
piRNA-88 | Downregulated | −2.78 |
piRNA-57 | Downregulated | −2.70 |
piRNA-76 | Downregulated | −2.45 |
piRNA-78 | Downregulated | −2.00 |
piRNA-77 | Downregulated | −1.91 |
piRNA-109 | Upregulated | 1.34 |
piRNA-20 | Upregulated | 1.96 |
piRNA-111 | Upregulated | 2.55 |
piRNA-112 | Upregulated | 4.13 |
Paclitaxel (nM) | KPT-330 (nM) | Fa Value | Combination Index (CI) Value |
---|---|---|---|
12.5 | 25 | 0.822 | 0.486 |
25 | 0 | 0.916 | 0.530 |
37.5 | 75 | 0.957 | 0.638 |
50 | 100 | 0.979 | 0.567 |
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Sexton, R.; Mahdi, Z.; Chaudhury, R.; Beydoun, R.; Aboukameel, A.; Khan, H.Y.; Baloglu, E.; Senapedis, W.; Landesman, Y.; Tesfaye, A.; et al. Targeting Nuclear Exporter Protein XPO1/CRM1 in Gastric Cancer. Int. J. Mol. Sci. 2019, 20, 4826. https://doi.org/10.3390/ijms20194826
Sexton R, Mahdi Z, Chaudhury R, Beydoun R, Aboukameel A, Khan HY, Baloglu E, Senapedis W, Landesman Y, Tesfaye A, et al. Targeting Nuclear Exporter Protein XPO1/CRM1 in Gastric Cancer. International Journal of Molecular Sciences. 2019; 20(19):4826. https://doi.org/10.3390/ijms20194826
Chicago/Turabian StyleSexton, Rachel, Zaid Mahdi, Rahman Chaudhury, Rafic Beydoun, Amro Aboukameel, Husain Y. Khan, Erkan Baloglu, William Senapedis, Yosef Landesman, Anteneh Tesfaye, and et al. 2019. "Targeting Nuclear Exporter Protein XPO1/CRM1 in Gastric Cancer" International Journal of Molecular Sciences 20, no. 19: 4826. https://doi.org/10.3390/ijms20194826