【摘要】 目的 探讨培美曲塞序贯吉非替尼对A549细胞的疗效,及其对A549细胞增殖、凋亡及凋亡相关蛋白表达的影响。方法 采用MTT[3-（4,5-二甲基噻唑-2）-2,5-二苯基四氮唑溴盐]比色法检测培美曲塞、吉非替尼、培美曲塞联合吉非替尼及培美曲塞序贯吉非替尼对A549细胞增殖的影响;Western blot检测凋亡相关蛋白的表达。DAPI核染色法检测A549细胞凋亡的形态学变化。结果 培美曲塞序贯吉非替尼对A549细胞增殖的抑制作用较单药及两药联合治疗更明显,A549细胞中培美曲塞、吉非替尼作用72 h的半数抑制浓度IC₅₀分别为0.84μmol/L和3.35μmol/L。Western blot检测到培美曲塞序贯吉非替尼组与培美曲塞、吉非替尼单药治疗组比较,Bcl-2、Caspase3、Caspase8及PARP蛋白表达明显降低,Bax、p-H2AX蛋白表达明显升高,差异有统计学意义（P<0.05）。DAPI核染色后可观察到培美曲塞序贯吉非替尼诱导A549细胞72 h后凋亡明显增强。结论 培美曲塞、吉非替尼均可明显抑制A549细胞增殖,诱导A549细胞凋亡,培美曲塞序贯吉非替尼作用更明显。更多还原

【Abstract】 Objective To investigate the effects of pemetrexed+gefitinib-gefitinib sequence on the proliferation and apoptosis of human non-small cell lung cancer（NSCLC） A549 cells.Methods The MTT method was used to detect the effect of pemetrexed（pemetrexed group）,gefitinib（gefitinib group）,combination of pemetrexed and gefitinib（pemetrexed+gefitinib group）,and pemetrexed+gefitinib-gefitinib sequence（pemetrexed+gefitinib-gefitinib group） on proliferation of A549 cells.Western blot was performed to analyze the expression of apoptosis-related proteins and the morphological changes of apoptosis were detected by DAPI staining.Results The inhibitory effects of pemetrexed or gefitinib alone and pemetrexed combined with gefitinib on A549 cells was observed.While the inhibitory effects of pemetrexed+gefitinib-gefitinib sequence were more obvious compared with pemetrexed or gefitinib alone and pemetrexed combined with gefitinib.The IC₅₀（inhibitory concentration 50%） values of pemetrexed and gefitinib on A549 cells for 72 h were 0.84 μmol/L and 3.35 μmol/L,respectively.Western blot showed that compared with pemetrexed group and gefitinib group,the expression of Bcl-2,caspase 3,caspase 8 and procyclic acidic repetitive proteins（PARP） in pemetrexed+gefitinib-gefitinib group was decreased,but Bax and p-H2AX was increased（P <0.05）.DAPI staining showed that the apoptosis rate of pemetrexed+gefitinib-gefitinib group on A549 cells for 72 h was the highest.Conclusion Both pemetrexed and gefitinib have obvious inhibitory and apoptosis effect on A549 cells,with the most prominent effect in the pemetrexed+gefitinib-gefitinib group.更多还原