活性氧在导致吉非替尼耐药细胞凋亡中的作用

Role of Reactive Oxygen Species in Apoptosis of Primary Gefitinib-Resistant Cancer Cells

目的:研究针对K-ras突变小分子NSC-741909是否可特异性杀伤吉非替尼原发耐药细胞,并对活性氧(reactive oxygenspecies,ROS)在其中的作用进行探讨。方法:选取H322(K-ras野生)及A549(K-ras突变)人源非小细胞肺癌细胞系,观察吉非替尼对不同细胞系生长的影响;NSC-741909作用于吉非替尼耐药细胞,观察细胞增殖与凋亡的变化;DCFH-DA荧光探针标记细胞内ROS,FCM检测细胞内ROS水平的变化,Western blot检测细胞内JNK通路蛋白变化。结果:K-ras突变型细胞对吉非替尼原发耐药,但NSC-741909可使这种原发耐药细胞出现凋亡;细胞内产生ROS,p-JNK表达增加而总JNK无改变,MKP1表达受抑制。结论:针对K-ras突变的小分子NSC-741909可使吉非替尼耐药细胞产生凋亡,NSC-741909通过使细胞内产生ROS持续激活JNK通路,是其导致吉非替尼耐药细胞凋亡的可能机制之一。

Objective： To investigate the specific cytotoxicity of the anticancer agent NSC-741909 on the growth of a non-small cell lung cancer （ NSCLC ） cell line primarily resistant to gefitinib, and to explore the role of reactive oxygen species （ ROS ） in the process. Methods： Two NSCLC cell lines with different sensitivities to gefitinib were selected, and the effects of gefltinib on cell growth, and induction of apoptosis induction by NSC-741909 were observed. The fluorescent probe 2＇,7＇-dichlorofluorescein diacetate （ DCFH-DA ） was used to label the intracellular ROS and the intracellular fluorescence intensity was detected using flow cytometry （ FCM ）. Jurl N-terminal kinase （ JNK ） activation was detected using Western blot analysis. Results： NSC-741909 induced the apoptosis of the NSCLC cells primarily resistant to gefltinib. ROS was detected in this cell line. There was an increase in P-JNK expression, whereas the total JNK protein expression remained unchanged. The MKP1 expression was suppressed. Conclusion： NSC-741909, which suppresses mutant K-Ras expression, induced the apoptosis of NSCLC cells that are primarily resistant to gefltinib. This inhibition was mediated by increased ROS production and subsequent JNK activation.