Sequence-dependent synergistic cytotoxicity of icotinib and pemetrexed in human lung cancer cell lines in vitro and in vivo
Sequence-dependent synergistic cytotoxicity of icotinib and pemetrexed in human lung cancer cell lines in vitro and in vivo
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DOI：
10.1186/s13046-019-1133-z
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发表时间：
2019-04-05
影响因子：
11.3
通讯作者：
Chen, Hongtao
中科院分区：
文献类型：
--
作者：
Liu, Tianze;Jin, Lizi;Chen, Hongtao
背景资料：近期的临床试验显示，表皮生长因子受体酪氨酸激酶抑制剂（EGFR-TKI）联合标准一线化疗未能改善晚期NSCLC患者的生存率，但EGFR-TKI联合化疗序贯治疗有望改善NSCLC患者的生存率。本研究旨在探讨培美曲塞与埃克替尼不同序贯联合应用对非小细胞肺癌（NSCLC）细胞株的抗增殖作用，以确定最佳联合方案，并探讨其可能的作用机制。采用CCK-8试剂盒和集落形成实验检测细胞增殖;流式细胞仪检测细胞周期和凋亡;创伤愈合实验检测细胞迁移和侵袭能力; Western blot检测蛋白表达。培美曲塞联合埃克替尼治疗（P-I）的抗癌能力显著强于埃克替尼联合培美曲塞治疗（I-P）和培美曲塞和埃克替尼联合治疗（P + I）。细胞周期分析显示培美曲塞阻滞细胞于S期，而埃克替尼阻滞细胞于G1期。我们还发现埃克替尼通过细胞色素C/Caspase/Bcl-2信号通路显著增强培美曲塞的促凋亡活性。此外，我们的研究结果表明，培美曲塞单独使用可增加p-EGFR、p-AKT和p-MAPK的水平，而埃克替尼可抑制这些水平。结论：培美曲塞联合埃克替尼序贯治疗NSCLC细胞具有较强的抗增殖作用，有望成为治疗NSCLC的一种新的有效联合方案。
Background: Recent Clinical trials of administration of epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) in combination with standard first-line chemotherapy have failed to improve survival in patients with advanced NSCLC, However, the sequential treatment with EGFR-TKIs and chemotherapy is expected to improve survival of NSCLC. The aim of this study is to test the antiproliferative effect of pemetrexed combined with icotinib in different sequences on non-small cell lung cancer (NSCLC) cell lines to determine the optimal combination schedule, and subsequently elaborated the potential mechanisms.Methods: Six human lung cancer cell lines with wild-type or mutant EGFR gene were exposed to pemetrexed and icotinib combined in different sequences. Cell proliferation was examined by cell counting kit-8 (CCK-8) and colony formation assay; cell cycle and apoptosis were evaluated by flow cytometry; cell migration and invasion were measured by wound healing and transwell invasion assays respectively; protein expression was by detected by Western blot.Results: The growth inhibition effect of pemetrexed combined with icotinib on NSCLC cells were schedule-dependent in vitro and in vivo. Treatment with pemetrexed followed by icotinib (P-I) had significantly stronger anticancer ability than treatment with icotinib followed by pemetrexed (I-P) and concomitant treatment with pemetrexed and icotinib (P + I). Cell cycle analysis revealed that pemetrexed blocked cells in S phase, whereas icotinib arrested cells in G1 phase. We also found that icotinib markedly enhanced the pro-apoptotic activity of pemetrexed via cytochrome-C/Caspase/Bcl-2 signaling pathway. In addition, our results showed that pemetrexed alone increased the levels of p-EGFR, p-AKT and p-MAPK, which were inhibited by icotinib. Finally, we showed that the washout period of icotinib was no less than 96 h.Conclusions: Sequential treatment of NSCLC cells with pemetrexed followed by icotinib had powerful antiproliferative effect, and it could become a novel effective combination therapy for NSCLC patients.