摘要
目的比较不同细胞株引入K-ras突变质粒前后对表皮生长因子受体（EGFR）信号传导通路活性的变化，探讨K—ras突变对于EGFR抑制剂耐药的机制。方法通过噻唑蓝（MTT）比色法检测HCC827及H292细胞在转染K-ras基因前后的EGFR下游AKT及STAT通路抑制剂敏感性变化，免疫印迹法（Western blot）检测EGFR下游通路激活状态变化。结果转染后HCC827细胞对于AKT和STAT通路抑制剂敏感性仅下降1．9和5．3倍，H292细胞对于AKT和STAT通路抑制剂敏感性仅下降3．0和2．5倍，远低于吉非替尼敏感性下降程度，转染K-ras的HCC827细胞AKT通路和STAT3通路持续激活，而转染K-ras的H292细胞AKT通路和STAT3通路的活性降低。结论K—ras导致EGFR抑制剂耐药的原因可能还存在其他耐药机制，携带EGFR基因突变或野生型的肺癌细胞中，K—ras突变对于AKT和STAT通路的影响可能存在不同。
Objective By comparing the activity of signaling pathway in different cell lines affect- ed by K-ras mutation, to investigate the resisitance to epidermal growth factor receptor （EGFR） inhibitors due to K-ras mutation. Methods Methyl thiazol tetrazolium （MTT） method was used to mensure the sen- sitivity and half maximal inhibitory concentration （ IC50 ） of AKT and STAT inhibitors after transfection of K-ras or blank plasmid into HCC827 and H292 cells. Western blotting was used to compare the activity of downstream signaling pathway in different cell lines. Results After transfection of mutant K-ras, the ICs0 of AKT and STAT inhibitors in HCC827 cells was reduced by 1.9 and 5.3 times respectively, and that in H292 cells was reduced by 3.0 and 2. 0 times respectively. The activity of AKT and STAT pathways in HCC827 cells transfected with mutant K-ras was continuously activated, while that in H292 cells was suppressed. Conclusion K-ras-induced resistance to EGFR inhibitors may be related to other mechanisms. In the tumor cells bearing mutant or wild-type EGFR gene, there may exist different influences of K-ras mutations on AKT and STAT pathways.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2011年第8期1232-1234,共3页
Chinese Journal of Experimental Surgery
基金
基金项目：国家自然科学基金资助项目（30700995）
北京市自然科学基金资助项目（7082097）
吴阶平基金会基金资助项目（08-2H-003）
关键词
表皮生长因子受体
K—ras
肺癌
耐药
Epidermal growth factor receptor
K-ras
Lung carcinoma
Resistance
