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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-smallcell 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subG1期细胞比例,并诱导细胞阻滞在G1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 investigate the inhibitory effect of sequential or combination of gefitinib (G) and paclitaxel (P) on non-small cell lung cancer (NSCLC) in three-dimensional cell model and its mechanism. METHODS: Human NSCLC cell lines A427 and Calu-3 were cultured in ultra low adhesion surface cell culture medium to form a three-dimensional cell model. The expression of paclitaxel was detected by sulforhodamine B (SRB) and Cell Titer-Blue Gefitinib (PG), gefitinib sequential paclitaxel (GP) and paclitaxel combined with gefitinib (P + G) on adherent and cultured three-dimensional cell proliferation inhibition, flow cytometry cells Cycle, Western blotting assay EGFR and Akt total protein and phosphorylation levels. Results: The survival rates of A427 cells were (39.5 ± 0.07)%, (57.7 ± 0.03)% and (53.7 ± 0.05)% when cultured in PG, GP and P + G cells, (23.9 ± 0.02)%, (58.2 ± 0.05)% and (48.8 ± 0.07)%, respectively. The inhibition of PG was the strongest (P <0.05). The PG, GP and P + The survival rates of A427 cells were (19.9 ± 2.89)%, (43.2 ± 8.64)% and (36.6 ± 9.79)%, respectively. The survival rates of Calu-3 cells were (10.2 ± 0.76)% and ± 3.45)% and (31.4 ± 6.15)%, respectively. The inhibitory effect of PG was the strongest (P <0.05). Moreover, the inhibitory effect of PG on these two cells was significantly stronger than that of monolayer cultured cells (P < 0.05). P-G treatment increased the percentage of subG1-phase cells and induced cell arrest in G1 phase. P-G treatment significantly decreased the levels of phosphorylated Akt and phosphorylated EGFR in three-dimensional cultured cells. CONCLUSION: In the three-dimensional cell model, P-G is more potent than P + G or G-P on the proliferation of NSCLC cells, which may be related to the cell cycle arrest and the down-regulation of phosphorylated Akt and EGFR.