Objective: To investigate the inhibitory effect of aumolertinib combined with anlotinib on proliferation of non-small cell lung cancer (NSCLC) cells.
Methods: CCK-8 assay, colony formation assay, and flow cytometry were used to assess the effect of different concentrations of aumolertinib or anlotinib on proliferation, survival, and apoptosis of PC-9 and HCC827 cells, and their synergistic effect was evaluated using the SynergyFinder model. In PC-9 and HCC827 cells treated with aumolertinib combined with anlotinib, the changes in cell invasion and migration abilities were assessed with Transwell assay, and the expressions of apoptosis- and invasion/migration-related proteins (Bax, Bcl-2, E-cadherin, vimentin, MMP2, and MMP9) and the key PI3K-Akt pathway proteins were detected using Western blotting.
Results: In PC-9 cells, the IC50 of aumolertinib and anlotinib was 1.701 μmol/L and 4.979 μmol/L, respectively, with a synergy score (ZIP) of 19.112; in HCC827 cells, their IC50 was 2.961 μmol/L and 7.934 μmol/L, respectively, with a ZIP of 12.325. Compared with aumolertinib and anlotinib used alone, their combined treatment more strongly inhibited the proliferation and survival, enhanced apoptosis and suppressed invasion and migration abilities of PC-9 and HCC827 cells. Western blotting showed that in both PC-9 and HCC827 cells, the combined treatment significantly upregulated the expressions of E-cadherin and Bax proteins, downregulated the expressions of Bcl-2, vimentin, MMP2, and MMP9 proteins, and reduced phosphorylation levels of PI3K and Akt.
Conclusion: Aumolertinib combined with anlotinib can effectively inhibit NSCLC cell proliferation by downregulating the PI3K-Akt pathway, suggesting a potentially new option for NSCLC treatment.
目的: 探究阿美替尼联合安罗替尼抑制非小细胞肺癌的作用。
方法: CCK-8法、集落克隆法和流式细胞术检测不同浓度阿美替尼和安罗替尼对PC-9细胞和HCC827细胞增殖、细胞存活和细胞凋亡的影响；SynergyFinder模型评价阿美替尼与安罗替尼的协同作用；Transwell小室实验检测阿美替尼联合安罗替尼对PC-9细胞和HCC827细胞侵袭及迁移的作用；Western blotting实验检测阿美替尼联合安罗替尼对PC-9细胞和HCC827细胞凋亡和侵袭迁移相关蛋白Bax、Bcl-2、E-Cadherin、Vimentin、MMP2和MMP9及PI3K-Akt通路关键蛋白表达的影响。
结果: 阿美替尼作用于PC-9细胞的IC50为1.701 μmol/L，安罗替尼作用于PC-9细胞的IC50=4.979 μmol/L，协同得分（ZIP）为19.112；阿美替尼对HCC827细胞IC50=2.961 μmol/L，安罗替尼对HCC827细胞IC50=7.934 μmol/L，协同得分（ZIP）为12.325，阿美替尼联合安罗替尼能够显著抑制PC-9细胞和HCC827细胞增殖（P<0.05）。相比于单药组，集落克隆结果表明阿美替尼联合安罗替尼能够显著抑制PC-9细胞和HCC827细胞存活（P<0.01）；流式细胞术结果表明，阿美替尼联合安罗替尼能够促进PC-9细胞和HCC827细胞发生凋亡（P<0.05）；Transwell实验结果表明，阿美替尼联合安罗替尼能够显著抑制PC-9细胞和HCC827细胞的侵袭迁移能力（P<0.01）；Western blotting实验结果表明，阿美替尼联合安罗替尼能够显著促进PC-9细胞和HCC827细胞中E-Cadherin和Bax蛋白的表达，抑制Bcl-2、Vimentin、MMP2和MMP9蛋白的表达（P<0.01），同时显著降低了PI3K-Akt通路中关键蛋白PI3K、Akt的磷酸化蛋白水平（P<0.01）。
结论: 阿美替尼联合安罗替尼通过下调PI3K-Akt通路抑制NSCLC肿瘤细胞，这种联合治疗方式可能成为临床上治疗 NSCLC患者的一种潜在治疗策略。
Keywords: PI3K-Akt pathway; anlotinib; apoptosis; aumolertinib; invasion and migration; non-small cell lung cancer.