吉非替尼对非小细胞肺癌细胞株H358增殖的影响及其作用机制

Effects and mechanism of gefitinib on the proliferation of non-small cell lung cancer cell line H358

目的观察吉非替尼(gefitinib)对非小细胞肺癌(NSCLC)细胞株H358增殖的影响,并对其分子机制进行探讨。方法体外培养人NSCLC细胞株H358,随机分为正常培养对照组和1μmol/L吉非替尼处理组,培养1、3、5d后,应用MTS法对2组细胞增殖速率进行分析;分别提取胞质、胞核蛋白,蛋白印迹法检测表皮生长因子受体(EGFR)的亚细胞定位变化情况;另外,对细胞中磷酸化-Akt(p-Akt)的表达进行检测,分析其对磷脂酰肌醇-3激酶(PI3K)/Akt信号通路的影响。结果 MTS结果表明,1μmol/L吉非替尼能显著抑制H358的增殖(P<0.01),并具有时间依赖性。蛋白印迹法检测结果显示,吉非替尼能够改变EGFR的亚细胞定位,减少其在胞核中的表达,此外,吉非替尼处理后H358细胞内p-Akt的蛋白表达受到抑制。结论吉非替尼能够显著抑制H358细胞的增殖,减少EGFR在细胞核中的表达,并抑制PI3K/Akt信号通路,这可能是其抗NSCLC的重要细胞及分子机制。

Objective To investigate the effects of gefitinib on the proliferation of non-small cell lung canc-er （NSCLC） cell line H358 ,and to explore the underlying molecular mechanism .Methods Human NSCLC H358 cells were cultured in vitro .Cells were divided randomly into the control group and gefitinib treated group ,after cultured for 1 ,3 ,5 days .The proliferation of H358 was investigated by M TS assay ,and the Western blotting was applied to investigate the sub-cellular location of epidermal growth factor receptor （EGFR） by extracting the protein of cytoplasm and nucleus separately ,and the protein expression of phospho-Akt ,a component of the PI3K/Akt signaling pathway .Results M TS assay results showed that gefitinib inhibited the proliferation of H 358 sig-nificantly （P〈0.01） in a time dependent manner .Western blotting results showed that the sub-cellular location of EGFR was affected by gefitinib and its expression in nucleus is reduced ,the protein expression of phospho-Akt in H358 cells was reduced by gefitinib .Conclusion Gefitinib inhibits the proliferation of H358 cells significantly , reduces the protein expression of EGFR in nucleus and inhibits the PI3K/Akt signaling pathway ,which may serve as an important mechanism for its anti-NSCLC effects .