目的建立人非小细胞肺癌吉非替尼( Gefitinib)耐药的细胞株PC9/Gefitinib,并研究其可能的耐药机制。方法采用体外浓度梯度递增的诱导方法建立Gefitinib获得性耐药细胞株 PC9/Gefitinib；MTT 法计算出 PC9和 PC9/Ge-fitinib的半数抑制浓度( IC50)和耐药指数( RI )及 PC9/Ge-fitinib细胞对多西他赛、培美曲塞及吉西他滨的敏感性；流式细胞仪检测其细胞周期分布；Western blot法检测p-AKT和Bcl-2蛋白的表达。结果建立了对 Gefitinib耐药的 PC9/Gefitinib细胞株,其RI是亲代PC9的106.95倍,且PC9/Ge-fitinib细胞株对化疗药物多西他赛的敏感性增加,而对培美曲塞表现出轻度耐药。耐药前后2种细胞株的G0/G1期、S期差异有统计学意义(P<0.05),G2/M期差异无统计学意义。与PC9细胞相比,PC9/Gefitinib中Bcl-2蛋白表达升高,而p-AKT表达水平下降。结论成功建立一株耐Gefitinib的人非小细胞肺癌细胞株PC9/Gefitinib,推测其耐药机制与Bcl-2蛋白表达升高及p-AKT表达下降有关。%Objective To establish PC9 cell line resistant to gefitinib and investigate its possible resistant mecha-nisms. Methods Human gefitinib-resistant non-small cell lung cancer cell line PC9/Gefitinib was established by gradually increasing concentration of gefitinib from its parental cell line PC9 in vitro. The IC50 value and resistance index( RI) of gefitinib were determined by MTT. MTT was also used to evaluate the sensitivity of docetaxel, peme-trexed and gemcitabine in PC9/Gefitinib cell line. Cell cycles were assayed by flow cytometry. Western blot analy-sis was used to investigate the expression of p-AKT and Bcl-2 . Results Gefitinib-resistant human non-small cell lung cancer PC9/Gefitinib had been established successfully, with the RI being 106.95. Compared to PC9 cells, PC9/Gefitinib cells had an increased sensitivity to docetaxel and showed certain resistance to pemetrexed. The cell cycles of G0/G1 period and S period had a significant difference between these two cell lines(P<0.05),the G2/M period change was not obvious. Compared with its parental PC9 cell line, the expression of Bcl-2 was significantly increased while the levels of p-AKT were obviously decreased in PC9/Gefitinib cell line. Conclusion Human ge-fitinib-resistant non-small cell lung cancer cell line PC9/Gefitinib is established successfully, and we speculate that the activation of Bcl-2 and decrease of p-AKT expression appeared to be associated with resistance to gefitinib.
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