Effect of the Knockout of FGL1 by CRISPR on the GefitinibSensitivity of PC9/GR Cells

Authors

  • Yufan Ji

DOI:

https://doi.org/10.61173/sbq56197

Keywords:

FGL1, CRISPR, PC9/GR, Gefitinib, Sensitivity

Abstract

This paper will investigate the effect of the Knockout of FGL1 by CRISPR on the EGFR signaling analyzed phosphor
ERK western blot and how it may control the gefitinib sensitivity of PC9/GR cells measured by the MTT Assay. This
experiment will knock out FGL1 using CRISPR. The Knockout will then be verified through Phospho-ERK western
blot. Moreover, the sensitivity of the gefitinib will be measured by the MTT Assay. The possible results generated from
the reports’ experiments may provide insight into future clinical trials of FGL1 Knockout consequences on gefitinib
sensitivity. Gefitinib is a potential drug for lung cancer. Therefore, investigating this drug may help scientists better
understand how the FGL1 may increase the sensitivity of Gefitinib cells and potentially discover a breakthrough to cure
cancer.

References

[1] Lee, J. H., Lee, D., Lu, M. T., Raghu, V. K., Park, C. M., Goo, J. M., ... & Kim, H. (2022). Deep Learning to Optimize Candidate Selection for Lung Cancer CT Screening: Advancing the 2021 USPSTF Recommendations. Radiology, 212877.

[2] Ettinger, D. S., Wood, D. E., Aisner, D. L., Akerley, W., Bauman, J. R., Bharat, A., Bruno, D. S., Chang, J. Y., Chirieac, L. R., D’Amico, T. A., Dilling, T. J., Dowell, J., Gettinger, S., Gubens, M. A., Hegde, A., Hennon, M., Lackner, R. P., Lanuti, M., Leal, T. A., ... Hughes, M. (2021). Non-small cell lung cancer, Version 2.2021 featured updates to the NCCN guidelines. JNCCN Journal of the National Comprehensive Cancer Network, 19(3), 254-266. https://doi.org/10.6004/jnccn.2021.0013

[3] Maemondo M, Inoue A, Kobayashi K, Sugawara S, Oizumi S, Isobe H, et al. Gefitinib or chemotherapy for non-small-cell lung cancer with mutated EGFR. N Engl J Med. 2010;362:2380–8.

[4] Shi, A. P., Tang, X. Y., Xiong, Y. L., Zheng, K. F., Liu, Y. Dean&Francis J., Shi, X. G., Lv, Y., Jiang, T., Ma, N., & Zhao, J. B. (2022). Immune Checkpoint LAG3 and Its Ligand FGL1 in Cancer. Frontiers in immunology, 12, 785091. https://doi.org/10.3389/ fimmu.2021.785091

[5] Wu, M., Yuan, Y., Pan, Y. Y., & Zhang, Y. (2014). Combined gefitinib and pemetrexed overcome the acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors in non-small cell lung cancer. Molecular medicine reports, 10(2), 931–938. https://doi.org/10.3892/mmr.2014.2243

[6] Yu, J., Li, J., Shen, J., Du, F., Wu, X., Li, M., Chen, Y., Cho, C. H., Li, X., Xiao, Z., & Zhao, Y. (2021). The role of Fibrinogenlike proteins in Cancer. International journal of biological sciences, 17(4), 1079–1087. https://doi.org/10.7150/ijbs.56748

[7] Song, YA., Ma, T., Zhang, XY. et al. Apatinib preferentially inhibits PC9 gefitinib-resistant cancer cells by inducing cell cycle arrest and inhibiting VEGFR signaling pathway. Cancer Cell Int 19, 117 (2019). https://doi.org/10.1186/s12935-019- 0836-8

[8] Roberts, P., Der, C. Targeting the Raf-MEK-ERK mitogenactivated protein kinase cascade for the treatment of cancer. Oncogene 26, 3291–3310 (2007). https://doi.org/10.1038/ sj.onc.1210422

[9] Miyamoto, Y., Suyama, K., & Baba, H. (2017). Recent Advances in Targeting the EGFR Signaling Pathway for the Treatment of Metastatic Colorectal Cancer. International Journal of Molecular Sciences, 18(4), 752. MDPI AG. Retrieved from http://dx.doi.org/10.3390/ijms18040752

[10] Agarwal, A., Mahfouz, R.Z., Sharma, R.K. et al. Potential biological role of poly (ADP-ribose) polymerase (PARP) in male gametes. Reprod Biol Endocrinol 7, 143 (2009). https://doi. org/10.1186/1477-7827-7-143

[11] Tanno, S., Ohsaki, Y., Nakanishi, K., Toyoshima, E., & Kikuchi, K. (2004). Small cell lung cancer cells express EGFR and tyrosine phosphorylation of EGFR is inhibited by gefitinib (‘Iressa’, ZD1839). Oncology Reports, 12, 1053-1057. https:// doi.org/10.3892/or.12.5.1053

[12] Sun, C., Gao, W., Liu, J., Cheng, H., & Hao, J. (2020). FGL1 regulates acquired resistance to Gefitinib by inhibiting apoptosis in non-small cell lung cancer. Respiratory research, 21(1), 210. https://doi.org/10.1186/s12931-020-01477-y

[13] Golding, S. E., Morgan, R. N., Adams, B. R., Hawkins, A. J., Povirk, L. F., & Valerie, K. (2009). Pro-survival AKT and ERK signaling from EGFR and mutant EGFRvIII enhances DNA double-strand break repair in human glioma cells. Cancer biology & therapy, 8(8), 730–738. https://doi.org/10.4161/ cbt.8.8.7927

[14] Changjiang Xu, Xiaoling Yuan, Zui Pan, Guoxiang Shen, Jung-Hwan Kim, Siwang Yu, Tin Oo Khor, Wenge Li, Jianjie Ma, Ah-Ng Tony Kong; Mechanism of action of isothiocyanates: the induction of ARE-regulated genes is associated with activation of ERK and JNK and the phosphorylation and nuclear translocation of Nrf2. Mol Cancer Ther 1 August 2006; 5 (8): 1918–1926. https://doi.org/10.1158/1535-7163.MCT-05-0497

[15] Noro, R., Gemma, A., Kosaihira, S., Kokubo, Y., Chen, M., Seike, M., Kataoka, K., Matsuda, K., Okano, T., Minegishi, Y., Yoshimura, A., & Kudoh, S. (2006). Gefitinib (IRESSA) sensitive lung cancer cell lines show phosphorylation of Akt without ligand stimulation. BMC cancer, 6, 277. https://doi. org/10.1186/1471-2407-6-277

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Published

2023-06-01