miR-196a Upregulation Contributes to Gefitinib Resistance through Inhibiting GLTP Expression
Abstract
:1. Introduction
2. Results
2.1. miR-196a Expression Levels Were Greatly Upregulated in Gefitinib-Resistant Lung Cancer Cells, and Its Levels Were Positively Correlated with Poor Overall Survival in Lung Cancer Patients
2.2. Forced Expression of miR-196a-Induced Gefitinib Resistance and Cell Proliferation, and Inhibited Cell Apoptosis That Was Important in Cell Resistance to Gefitinib Treatment
2.3. NRF2 Expression Levels Were Increased in Gefitinib-Resistant Cells and NRF2 Induced miR-196a Expression at Transcriptional Level
2.4. GLTP Was a Direct Functional Target of miR-196a, and GLTP Suppression Was Important for the Resistance of Cells to Gefitinib Treatment
2.5. GLTP-Forced Expression Inhibited miR-196a-Inducing Gefitinib Resistance
2.6. miR-196a-Forced Expression Induced Tumor Growth and Inhibited Cell Apoptosis In Vivo, Which May Contribute to Gefitinib Resistance
3. Discussion
4. Materials and Methods
4.1. Human Lung Cancer Specimens
4.2. Cell Culture and Reagents
4.3. RNA Extraction and Quantitative Real-Time PCR Analysis
4.4. Cell Viability Assay
4.5. Cell Apoptosis Assay
4.6. Western Blot Analysis
4.7. Transfection
4.8. Luciferase Reporter Assay
4.9. Tumor Growth Assay In Vivo
4.10. Immunohistochemistry (IHC) Assay
4.11. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Liu, B.-J.; Li, F.-F.; Xie, Y.-X.; Fan, C.-Y.; Liu, W.-J.; Qiu, J.-G.; Jiang, B.-H. miR-196a Upregulation Contributes to Gefitinib Resistance through Inhibiting GLTP Expression. Int. J. Mol. Sci. 2022, 23, 1785. https://doi.org/10.3390/ijms23031785
Liu B-J, Li F-F, Xie Y-X, Fan C-Y, Liu W-J, Qiu J-G, Jiang B-H. miR-196a Upregulation Contributes to Gefitinib Resistance through Inhibiting GLTP Expression. International Journal of Molecular Sciences. 2022; 23(3):1785. https://doi.org/10.3390/ijms23031785
Chicago/Turabian StyleLiu, Bing-Jie, Fang-Fang Li, Yun-Xia Xie, Chong-Yuan Fan, Wen-Jing Liu, Jian-Ge Qiu, and Bing-Hua Jiang. 2022. "miR-196a Upregulation Contributes to Gefitinib Resistance through Inhibiting GLTP Expression" International Journal of Molecular Sciences 23, no. 3: 1785. https://doi.org/10.3390/ijms23031785