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Macau Periodical Index (澳門期刊論文索引)

Author
Yu, Dian；Yin, Tianpeng；Li, Gang；Wang, Caiyun；Bai, Li-ping；Zhu, Guoyuan；Zhang, Wei
Title
The Combination of aloe vera and Gefitinib effectively suppresses growth and migration of Gefitinib-resistant H1975 lung cancer cells and inhibits Wnt/β-catenin signaling
Journal Name
澳門科技大學學報
Pub. Info
2023年9月30日, 第17卷第3期, pp. 79-107
Link
https://www.mustjournal.com/CN/10.58664/mustjournal.2023.09.005
Keyword
Aloe vera；Gefitinib；Gefitinib-resistant；EGFR T790M；β-catenin
Abstract
Lung cancer treatment has been revolutionized by the discovery of somatic mutations in epidermal growth factor receptors (EGFRs) and the development of EGFR tyrosine kinase inhibitors (TKIs). However, in most cases, non-small cell lung cancer (NSCLC) patients who are treated with gefitinib eventually develop resistance. The purpose of this study was to investigate the effect of Aloe vera (L.) Burm.f. (A. vera) extract combined with gefitinib on gefitinib-resistant NSCLC cell lines. The cytotoxicity assay showed that there was a minimal inhibitory effect of A. vera on cell proliferation in A549, PC-9, and H1975 cells, while the combined use of A. vera and gefitinib resulted in a significant inhibitory effect on cell growth in a concentration-dependent manner. Importantly, compared to other cell lines, H1975 proliferation was inhibited by the combined treatment more effectively. The IC50 values at 24 h decreased from 37.92 ± 1.19 to 20.09 ± 1.03 μM for H1975 cells. As a result of the combined treatment of A. vera and gefitinib, the migration ability of H1975 cells was decreased compared with that of the gefitinib group. Moreover, A. vera significantly increased the gefitinib-induced apoptosis in H1975 cells and the combination treatment induced G2/M phase arrest in H1975 cells. Furthermore, the Wnt/β-catenin pathway was inhibited in H1975 cells of the high-concentration combination group, suggesting that A. vera is a potential anticancer therapeutic adjuvant for gefitinib-resistant NSCLC with the EGFR-T790M mutation. Paragraph Headings: 1. Introduction 2. Materials and methods 2.1. A. vera preparation 2.2. Cell lines and cell culture 2.3. Sulforhodamine B assay for cytotoxicity screening 2.4. Scratch wound healing assay 2.5. Apoptosis assay 2.6. Cell cycle analysis 2.7. Western blot analysis 2.8. Statistical analysis 3. Results 3.1. Combination treatment with A. vera and gefitinib inhibits NSCLC cell growth 3.2. Combination treatment with A. vera and gefitinib decreases the migration ability of H1975 cells 3.3. A. vera enhances gefitinib-induced apoptosis in H1975 cells 3.4. Combination of A.vera and gefitinib induced G2/M phase arrest in H1975 cells 3.5. Combination of A. vera and gefitinib inhibits the Wnt/β-catenin signaling in H1975 cells 4. Discussion 5. Conclusion Tables: 1. Characteristics of NSCLC cell lines used in the study Figures: 1. Cytotoxic effects of gefitinib and A. vera on NSCLC cells: (a) Cells were incubated with various doses of A.vera for 24 h and cell viability was analyzed using an SRB assay; (b) H1975 cells were treated with A. vera (200 μg/mL) combined with different doses of gefitinib for 24 h. Cell viability was examined by SRB assay and absorbance was read at 570 nm. The error bars, represent the standard deviation of three independent measurements. 2. A. vera and gefitinib inhibit cell migration in gefitinib-resistant H1975 cells for 24 h. (a) Scratch wound healing assay. (b) Changes in cell motility were assessed by scratch wound healing assay. Error bars indicate the standard deviation of three independent measurements. (∗ p < 0.05, ∗∗ p < 0.01, compared with the Gefitinib group). 3. A. vera and gefitinib induce the cell apoptosis in gefitinib-resistant H1975 cells for 24h. (a) Representative images of cell apoptosis analysis after the indicated treatment. (b) The graph indicated the average ratio (column) ± SD of cell apoptosis (∗ p < 0.05, ∗∗ p < 0.01, compared with Gefitinib group). 4. A. vera and gefitinib induce the G2/M phase cell cycle arrest in gefitinib-resistant H1975 cells for 24 h. (a) Representative images of cell cycle analysis after the indicated treatment in H1975 cells. (b) The graph demonstrated the average ratio (column) ± SD at different phases. (∗ p < 0.05, compared with Gefitinib group). 5. Detection of the Wnt/β-catenin pathway in H1975 cells. (a) The levels of Wnt/β-catenin pathway proteins were assessed by Western blotting, and GAPDH was used as a loading control. (b) Relative expression levels of Wnt/β-catenin pathway proteins. (∗ p < 0.05, ∗∗ p < 0.01, compared with the Gefitinib group).