目的： 探讨序贯或联合吉非替尼（gefitinib，G）和紫杉醇（paclitaxel，P）对三维细胞模型中非小细胞肺癌（non-small cell lung cancer，NSCLC）的抑制作用及其机制。 方法: 采用超低黏附表面细胞培养板培养人NSCLC细胞株A427、Calu-3，使其形成三维细胞模型；分别采用磺酸罗丹明B（sulforhodamine B，SRB）、Cell Titer-Blue法检测紫杉醇序贯吉非替尼（P-G）、吉非替尼序贯紫杉醇（G-P）及紫杉醇联合吉非替尼（P+G）处理对贴壁和三维培养细胞增殖的抑制作用，流式细胞术检测细胞周期，Western blotting检测细胞中EGFR和Akt总蛋白及其磷酸化的水平。 结果: 单层细胞培养时，P-G、G-P和P+G处理后，A427细胞的存活率分别为（39.5±0.07）%、（57.7±0.03）%和（53.7±0.05）%，Calu-3细胞的存活率分别为（23.9±002）%、（58.2±0.05）%和（48.8±0.07）%，以P-G的抑制作用最强（P<0.05）；三维细胞模型中，P-G、G-P和P+G处理后，A427细胞的存活率分别为（19.9±2.89）%、（43.2±8.64）%和（36.6±9.79）%，Calu-3细胞的存活率分别为（10.2±076）%、（50.0±3.45）%和（31.4±6.15）%，也以P-G的抑制作用最强（P<0.05）；而且，P-G对这两细胞的抑制作用，三维培养细胞显著强于单层培养细胞（P<0.05）。P-G治疗可提高subG1期细胞比例，并诱导细胞阻滞在G1期；P-G治疗可明显下调三维培养细胞中磷酸化Akt和磷酸化EGFR的水平。 结论: 三维细胞模型中，P-G较P+G或G-P对NSCLC细胞的增殖有更强的抑制作用，可能与细胞周期阻滞和磷酸化Akt、EGFR水平下调有关。

Objective：To explore the inhibitory effect and mechanism of sequential or combined treatment of gefitinib (G) and paclitaxel (P) on three-dimensional non-small cell lung cancer (NSCLC) cell lines. Methods: The human NSCLC cell lines A427 and Calu-3 were cultured in the ultralow attachment plates and the three-dimensional cell clusters were formed; the inhibitory effect of paclitaxel followed by gefitinib (P-G), gefitinib followed by paclitaxel (G-P), and paclitaxel combined with gefitinib (P+G) on the monolayer and three-dimensional cell lines were detected using sulforhodamine B (SRB) and Cell Titer-Blue assay, respectively; the cell cycle was detected by flow cytometry; and the expression of total EGFR and Akt protein and their phosphorylation were detected using Western blotting. Results: In the monolayer cells, the survival rates of A427 cells in the group of P-G, G-P, and P+G were (39.5±0.07)%, (57.7±003)% and (53.7±0.05)% respectively. Those of Calu-3 cells in the group of P-G, G-P and P+G were (23.9±002)%, (58.2±0.05)% and (48.8±0.07)% respectively. The inhibitory effect of P-G was the strongest (P<005). In the three-dimensional cells, the survival rates of A427 cells in the group of P-G, G-P and P+G were (19.9±2.89)%, (43.2±8.64)% and (36.6±9.79)% respectively. Those of Calu-3 cells in the group of P-G, G-P and P+G were (10.2±0.76)%, (50.0±3.45)%, (31.4±6.15)% respectively. The inhibitory effect of P-G was the strongest (P<0.05). In addition, the inhibitory effects of P-G on A427 and Calu-3 in the three-dimensional cells were stronger than those in the monolayer cells (P<0.05). The sequence of P-G resulted in an increase of subG1 proportion and arrested monolayer cells in G1 phase. The phosphorylation of EGFR and Akt was decreased in subsequent exposure to P-G. Conclusion: The sequence of P-G seems to be superior to the reverse schedule or combination in the inhibition of proliferation of NSCLC cells, based on the factors of G1 arrest and the decrease of phosphorylation of Akt and EGFR in three-dimensional cells.

国家自然科学基金资助项目（No. 30873023）；同济大学青年优秀人才培养计划项目（No. 1511219011）