摘要
目的:探讨人髓细胞白血病基因-1(myeloid cell leukemia-1,Mcl-1)是否参与调控非小细胞肺癌(non-small cell lung cancer,NSCLC)对EGFR-TKIs的敏感性,为非小细胞肺癌的治疗提供新的思路。方法:通过Western blot方法检测EGFR-TKIs敏感细胞H3255和耐药细胞H1975中Mcl-1蛋白的表达水平。分别给予敏感细胞H3255和耐药细胞H1975 EGFR-TKIs处理后检测细胞凋亡情况和Mcl-1蛋白表达水平。设计并合成特异性si RNA下调耐药细胞H1975中Mcl-1的表达,采用脂质体转染后通过流式细胞技术检测细胞的凋亡情况。结果:H3255细胞Mcl-1表达水平明显低于H1975细胞。一代EGFR-TKIs Gefitinib显著降低H3255细胞Mcl-1表达而不能减少H1975细胞Mcl-1表达。H1975细胞经二代EGFR-TKIs Afatinib和三代EGFR-TKIs AZD9291处理后Mcl-1表达明显减少。特异性si RNA下调H1975细胞Mcl-1表达可以促进细胞凋亡。结论:Mcl-1参与了调节NSCLC对EGFR-TKIs的敏感性,可能成为防止或逆转NSCLC对EGFR-TKIs耐药的潜在靶点。
Objective： To explore whether myeloid cell leukemia-1 （Mcl-1） participated in regulating the sensitivity to epidermal growth factor receptor tyrosine kinase inhibitors of non-small cell lung cancer, and provide new ideas for the treatment of non -small cell lung cancer （NSCLC）. Methods： The expression levels of Mcl-1 in EGFR-TKIs sensitive cell H3255 and resistant cell H1975 were detected by western blot. Then the expression level of Mcl-1 and apoptosis of sensitive cell H3255 and resistant cell HI 975 after being treated with EGFR-TKIs were examined. The specific siRNA that could down-regulate the expression of Mcl-1 in resistant cell H1975 was designed and synthesized, and the apoptosis of these cells wer detected by flow cytometry after liposome transfection. Results： The expression of Mcl-1 in H3255 cell was significantly lower than that in H1975 cell. Meanwhile, the expression of Mcl-1 prominently diminished in H3255 cell and the expression of Mcl-1 in H1975 cell had no obvious change in the presence of first generation EGFR-TKIs Gefitinib. The expression of Mcl-1 was obviously decreased inH1975 cell after being treated with second generation EGFR-TKIs Afatinib and third generation EGFR-TKIs AZD9291. Flow cytometry showed that specific siRNA which down-regulated the expression of Mcl-1 in H1975 cell could promote the apoptosis. Conclusion： Mcl-1 participated in regulating the sensitivity to EGFR-TKIs of NSCLC, and might be regarded as a target to overcome the resistance to EGFR-TKIs of NSCLC.
出处
《现代生物医学进展》
CAS
2016年第16期3028-3031,3077,共5页
Progress in Modern Biomedicine
基金
国家自然科学基金项目(81272518)
