|
|
吉非替尼与TRAIL协同诱导A549人肺腺癌细胞系凋亡的作用研究
|
Abstract:
目的：本论文旨在探讨吉非替尼与肿瘤坏死因子相关凋亡诱导配体TRAIL (TNF-Related Apoptosis Inducing Ligand)的协同抗人肺癌作用并阐明两者协同的机制。方法：实验采用人A549肺癌细胞。实验分为对照组、吉非替尼处理组、TRAIL处理组及吉非替尼和TRAIL共同处理组。分别采用MTT实验，流式细胞术，Western-blot检测吉非替尼和TRAIL对A549细胞的存活、凋亡以及凋亡相关蛋白等方面的影响。结果：吉非替尼和TRAIL共同处理组的细胞存活率相较于空白对照组和各单独处理组均有显著变化。流式细胞术检测结果显示：对照组诱导的A549细胞凋亡比率分别为2.7%，15 μM吉非替尼单独处理组诱导的凋亡率为8.5%，100 ng/ml TRAIL单独处理组诱导的凋亡率为17.5%，而两者共同处理组诱导的凋亡比例为69.8%。相较与对照组或各单独处理组都有显著升高(P < 0.01)。Western-blot实验结果表明：吉非替尼和TRAIL显著诱导凋亡相关蛋白的水解。结论：吉非替尼和TRAIL可协同抑制肺癌细胞存活，促进细胞凋亡。
Objective: This paper aims to explore the synergistic anti-lung cancer effect of gefitinib and TRAIL (TNF-Related Apoptosis Inducing Ligand), and elucidate the mechanism of their synergy. Method: The experiment applied human A549 lung cancer cells. The experiment was divided into control group, gefitinib treatment group, TRAIL treatment group, and gefitinib and TRAIL co-treatment group. MTT assay, flow cytometry, and Western-blot were used to detect the effects of gefitinib and TRAIL on the cell viability, apoptosis, and apoptosis related proteins of A549 cells. Result: The cell viability of the co-treated group with gefitinib and TRAIL showed significant changes compared to the control group and each individual treatment group. The results of flow cytometry showed that the apoptosis ratio of A549 cells induced by the control group were 2.7%. The apoptosis induced by 15 μM of gefitinib alone treatment group was 8.5%, the apoptosis induced by 100 ng/ml TRAIL alone treatment group was 17.5%, and the apoptosis induced by the combined treatment group was 69.8%. Compared with the control group or each individual treatment group, there was a sig-nificant increase (P < 0.01). The Western-blot experiment results showed that gefitinib and TRAIL significantly induced the hydrolysis of apoptosis related proteins. Conclusion: Gefitinib and TRAIL can synergistically inhibit the viability of lung cancer cells and promote cell apoptosis.
| [1] | Ivanov, V.N., Bhoumik, A. and Ronai, Z. (2003) Death Receptors and Melanoma Resistance to Apoptosis. Oncogene, 22, 3152-3161. https://doi.org/10.1038/sj.onc.1206456 |
| [2] | Leblanc, H.N. and Ashkenazi, A. (2003) Apo2L/TRAIL and Its Death and Decoy Receptors. Cell Death & Differentiation, 10, 66-75. https://doi.org/10.1038/sj.cdd.4401187 |
| [3] | Ozoren, N. and El-Deiry, W.S. (2003) Cell Surface Death Receptor Signaling in Normal and Cancer Cells. Seminars in Cancer Biology, 13, 135-147. https://doi.org/10.1016/S1044-579X(02)00131-1 |
| [4] | Montinaro, A. and Walczak, H. (2023) Harnessing TRAIL-Induced Cell Death for Cancer Therapy: A Long Walk with Thrilling Discoveries. Cell Death & Differentiation, 30, 237-249. https://doi.org/10.1038/s41418-022-01059-z |
| [5] | Hellwig, C.T., Eugenia Delgado, M., Skoko, J., et al. (2022) Proteasome Inhibition Triggers the Formation of TRAIL Receptor 2 Platforms for Caspase-8 Activation That Accumulate in the Cytosol. Cell Death & Differentiation, 29, 147-155. https://doi.org/10.1038/s41418-021-00843-7 |
| [6] | Phillips, D.C., Buchanan, F.G., Cheng, D., Solomon, L.R., Xiao, Y., Xue, J., et al. (2021) Hexavalent TRAIL Fusion Protein Eftozanermin Alfa Optimally Clusters Apoptosis-Inducing TRAIL Receptors to Induce on-Target Antitumor Activity in Solid Tumors. Cancer Research, 81, 3402-3414. https://doi.org/10.1158/0008-5472.CAN-20-2178 |
| [7] | Overdijk, M.B., Strumane, K., Beurskens, F.J., Ortiz Buijsse, A., Vermot-Desroches, C., Vuillermoz, B.S., et al. (2020) Dual Epitope Targeting and Enhanced Hexamerization by DR5 Antibodies as a Novel Approach to Induce Potent Antitumor Activity through DR5 Agonism. Molecular Cancer Therapeutics, 19, 2126-2138.
https://doi.org/10.1158/1535-7163.MCT-20-0044 |
| [8] | Kumar, P., Mangla, B., Javed, S., et al. (2023) Gefitinib: An Updated Review of Its Role in the Cancer Management, Its Nanotechnological Interventions, Recent Patents and Clinical Trials. Recent Patents on Anti-Cancer Drug Discovery, 18, 448-469. https://doi.org/10.2174/1574892818666221026164940 |
| [9] | Lemke, J., von Karstedt, S., Zinngrebe, J. and Walczak, H. (2014) Getting TRAIL Back on Track for Cancer Therapy. Cell Death & Differentiation, 21, 1350-1364. https://doi.org/10.1038/cdd.2014.81 |
| [10] | Belada, D., Mayer, J., Czuczman, M.S., et al. (2010) Phase II Study of Dulanermin plus Rituximab in Patients with Relapsed Follicular Non-Hodgkin’s Lymphoma (NHL). Journal of Clinical Oncology, 28, Article 8104.
https://doi.org/10.1200/jco.2010.28.15_suppl.8104 |
| [11] | Cohen, S.J., Cohen, R.B. and Meropol, N.J. (2005) Tar-geting Signal Transduction Pathways in Colorectal Cancer—More than Skin Deep. Journal of Clinical Oncology, 23, 5374-5385. https://doi.org/10.1200/JCO.2005.02.194 |
| [12] | Yuan, Y., Li, X.F., Chen, J.Q., et al. (2014) Critical Appraisal of the Role of Gefitinib in the Management of Locally Advanced or Metastatic Non-Small Cell Lung Cancer. OncoTargets and Therapy, 28, 841-852.
https://doi.org/10.2147/OTT.S34124 |
| [13] | Yerbes, R., López-Rivas, A., Reginato, M.J. and Palacios, C. (2012) Control of FLIPL Expression and TRAIL Resistance by the Extracellular Signal-Regulated Kinase1/2 Pathway in Breast Epithelial Cells. Cell Death & Differentiation, 19, 1908-1916. https://doi.org/10.1038/cdd.2012.78 |
| [14] | Jun, H., Jang, E., Kim, H., et al. (2022) TRAIL & EGFR Affibody Dual-Display on a Protein Nanoparticle Synergistically Sup-presses Tumor Growth. Journal of Controlled Release, 349, 367-378.
https://doi.org/10.1016/j.jconrel.2022.07.004 |
