# 新型TK抑制剂达克替尼在肺腺癌细胞增殖和凋亡的影响Effect of a Novel TK Inhibitor Dacomitinib on Proliferation and Apoptosis of Lung Adenocarcinoma Cells

Abstract: Objective: The objective is to investigate the proliferation and invasion of human lung adenocar-cinoma A549 cells induced by a novel TK inhibitor, dacomitinib, and its mechanism. Methods: Human lung adenocarcinoma cell line A549 was divided into three groups according to the pre-liminary results, namely the blank control group, the low-dose group and the high-dose group. The expression of EFGR and Caspase-3 and Caspase-9 was detected by RT-PCR. The cycle of cell division and apoptotic rate of the cell were detected by flow cytometry. The cell proliferation was detected by CCK-8. Results: The relative expressions of EFGR, caspase-3 and caspase-9 mRNA in blank control group, low-dose group and high-dose group were statistically different (P < 0.05). The relative expression of EFGR mRNA in the blank control group was significantly higher than that in the low-dose group and the high-dose group (P < 0.05). The relative expression of Caspase-3 and caspase-9 mRNA in the blank control group was significantly lower than that in the low-dose group and high-dose group (P < 0.05). There was no significant difference in the relative expression of EFGR, caspase-3 and caspase-9 mRNA between the low-dose group and the high-dose group (P > 0.05). The proportions of G0/G1 phase in low-dose group and high-dose group were significantly higher than those in blank control group (P < 0.05), while the proportions of S phase and G2 phase were significantly lower than those in blank control group (P < 0.05). The total apoptotic rate of the low-dose group and the high-dose group significantly higher than that of the blank control group (P < 0.05), and the late apoptotic rate of the high-dose group was significantly higher than that of the low-dose group (P < 0.05). After 2 days of drug treatment, the cell increment rate of blank control group was significantly higher than that of low-dose group and high-dose group (P < 0.05); after 3 days of drug treatment, the cell increment rate of blank control group was significantly higher than that of low-dose group and high-dose group (P < 0.05). Conclusion: Low-dose of dacomitinib can inhibit the proliferation of lung adenocarcinoma cells, promote the apoptosis of lung adenocarcinoma cells, and inhibit the expression of EFGR. It can be used as a therapeutic regimen for drug-resistant lung adenocarcinoma.Objective: The objective is to investigate the proliferation and invasion of human lung adenocarci-noma A549 cells induced by a novel TK inhibitor, dacomitinib, and its mechanism. Methods: Human lung adenocarcinoma cell line A549 was divided into three groups according to the preliminary re-sults, namely the blank control group, the low-dose group and the high-dose group. The expression of EFGR and Caspase-3 and Caspase-9 was detected by RT-PCR. The cycle of cell division and apop-totic rate of the cell were detected by flow cytometry. The cell proliferation was detected by CCK-8. Results: The relative expressions of EFGR, caspase-3 and caspase-9 mRNA in blank control group, low-dose group and high-dose group were statistically different (P < 0.05). The relative expression of EFGR mRNA in the blank control group was significantly higher than that in the low-dose group and the high-dose group (P < 0.05). The relative expression of Caspase-3 and caspase-9 mRNA in the blank control group was significantly lower than that in the low-dose group and high-dose group (P < 0.05). There was no significant difference in the relative expression of EFGR, caspase-3 and caspa-se-9 mRNA between the low-dose group and the high-dose group (P > 0.05). The proportions of G0/G1 phase in low-dose group and high-dose group were significantly higher than those in blank control group (P < 0.05), while the proportions of S phase and G2 phase were significantly lower than those in blank control group (P < 0.05). The total apoptotic rate of the low-dose group and the high-dose group significantly higher than that of the blank control group (P < 0.05), and the late apoptotic rate of the high-dose group was significantly higher than that of the low-dose group (P < 0.05). After 2 days of drug treatment, the cell increment rate of blank control group was signifi-cantly higher than that of low-dose group and high-dose group (P < 0.05); after 3 days of drug treatment, the cell increment rate of blank control group was significantly higher than that of low-dose group and high-dose group (P < 0.05). Conclusion: Low-dose of dacomitinib can inhibit the proliferation of lung adenocarcinoma cells, promote the apoptosis of lung adenocarcinoma cells, and inhibit the expression of EFGR. It can be used as a therapeutic regimen for drug-resistant lung ade-nocarcinoma.

1. 引言

2. 材料与方法

2.1. 肺腺癌细胞系A549的培养

2.2. 实验分组

2.3. 实时荧光定量PCR检测Akt1、Caspase-3和Caspase-9 mRNA的表达水平

Table 1. The primer sequence of target gene

2.4. 细胞凋亡水平

2.5. 细胞增殖能力

2.6. 统计学方法

3. 结果

3.1. EFGR，Caspase-3和Caspase-9的mRNA相对表达量

Figure 1. The relative mRNA expression of EFGR, caspase-3 and caspase-9. Note: ** indicates statistical difference (P < 0.05); # indicates no statistical difference (P > 0.05)

3.2. 3组细胞周期比较结果

3.3. 3组细胞凋亡率比较结果

3.4. 3组细胞增殖率比较结果

(a) (b)(c)

Figure 2. The comparison of cell cycle among three groups. (a) The blank control group; (b) The low-dose group; (c) The high-dose group

(a) (b)(c)

Figure 3. The comparison of apoptosis rate among three groups. (a) The blank control group; (b) The low-dose group; (c) The high-dose group

Figure 4. The comparison of proliferation rate among three groups. Note: * indicates statistical difference (P < 0.05)

4. 讨论

[1] Jordan, E.J., Kim, H.R., Arcila, M.E., et al. (2017) Prospective Comprehensive Molecular Characterization of Lung Adenocarcinomas for Efficient Patient Matching to Approved and Emerging Therapies. Cancer Discovery, 7, 15 p.
https://doi.org/10.1158/2159-8290.CD-16-1337

[2] Velcheti, V., Hida, T., Reckamp, K.L., et al. (2017) Phase 2 Study of Lenvatinib in Patients with RET Fusion-Positive Adenocarcinoma of the Lung. European Journal of Cancer, 72, S178.
https://doi.org/10.1016/S0959-8049(17)30651-2

[3] Feng, M., Zhu, J., Liang, L., et al. (2017) Diagnostic Value of Tumor Markers for Lung Adenocarcinoma-Associated Malignant Pleural Effusion: A Validation Study and Me-ta-Analysis. International Journal of Clinical Oncology, 22, 283-290.
https://doi.org/10.1007/s10147-016-1073-y

[4] Ono, A., Kenmotsu, H., Watanabe, M., et al. (2014) Mutant Al-lele Frequency Predicts the Efficacy of EGFR-TKIs in Lung Adenocarcinoma Harboring the L858R Mutation. Annals of Oncology, 25, 1948-1953.
https://doi.org/10.1093/annonc/mdu251

[5] Kim, I.A., Lee, J.S., Kim, H.J., et al. (2018) Cumulative Smoking Dose Affects the Clinical Outcomes of EGFR-Mutated Lung Adenocarcinoma Patients Treated with EGFR-TKIs: A Retrospective Study. BMC Cancer, 18, Article No. 768.
https://doi.org/10.1186/s12885-018-4691-0

[6] Zhou, F., Ma, W., Li, W., et al. (2018) Thick-Wall Cavity Predicts Worse Progression-Free Survival in Lung Adenocarcinoma Treated with First-Line EGFR-TKIs. BMC Cancer, 18, Article No. 1033.
https://doi.org/10.1186/s12885-018-4938-9

[7] Chen, H., Yang, X., Liu, H., et al. (2017) Correlation between Serum Tumor Markers and Efficacy of First-Line EGFR-TKIs in Patients with Advanced Lung Adenocarcinoma. Chi-nese Journal of Lung Cancer, 1, 76-87.

[8] Ma, X., Zhu, H., Guo, H., et al. (2016) Risk Factors of Brain Metastasis during the Course of EGFR-TKIs Therapy for Patients with EGFR-Mutated Advanced Lung Adenocarcinoma. Onco-target, 7, 81906-81917.
https://doi.org/10.18632/oncotarget.11918

[9] Ming-Szu, H., Jr-Hau, L., Lin, Y.C., et al. (2016) The Content of Mutant EGFR DNA Correlates with Response to EGFR-TKIs in Lung Adenocarcinoma Patients with Common EGFR Mutations. Medicine, 95, e3991.
https://doi.org/10.1097/MD.0000000000003991

[10] 周玮玮, 王磊, 苗玉, 等. 量子点荧光探针检测肺腺癌组织中EGFR的表达[J]. 标记免疫分析与临床, 2018, 25(5): 736-739.

[11] 史张, 宋长恩, 施睿峰, 等. 肺腺癌CT及临床特征与EGFR 19号外显子突变的相关性研究[J]. 临床放射学杂志, 2017, 36(4): 490-494.

[12] Bose, T.O., Pham, Q.M., Jellison, E.R., et al. (2013) CD11a Regulates Effector CD8 T Cell Differentiation and Central Memory Development in Response to Infection with Listeria Monocytogenes. Infection and Immunity, 81, 1140-1151.
https://doi.org/10.1128/IAI.00749-12

[13] Tai, W., Chen, Z. and Cheng, K. (2013) Expression Profile and Func-tional Activity of Peptide Transporters in Prostate Cancer Cells. Molecular Pharmaceutics, 10, 477-487.
https://doi.org/10.1021/mp300364k

[14] Ferrone, S. and Marincola, F.M. (1995) Loss of HLA Class I Antigens by Melanoma Cells: Molecular Mechanisms, Functional Significance and Clinical Relevance. Immunology Today, 16, 487-494.
https://doi.org/10.1016/0167-5699(95)80033-6

Top