非小细胞肺癌K-ras突变小分子对吉非替尼耐药细胞的作用

目的 观察针对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,并探讨其机制.方法 选取吉非替尼耐药细胞;观察NSC-741909作用后细胞增殖与凋亡的变化;共聚焦显微镜观察细胞骨架改变;Western blot检测K-ras、JNK、P-JNK的改变.结果 NSC-741909可使耐药细胞在作用24 h后,在1 μmol/L和2 μmol/L时,FITC-A+/PE-A+细胞增加至(6.9±0.6)%和(21.1±3.2)%(P＜0.01);作用30 min后,K-ras表达在2 h下降达70%;p-JNK表达增加,并持续至少15 h,而总JNK蛋白表达无改变;细胞骨架蛋白F-actin呈稀疏、不规则、发散状排列.结论 针对K-ras突变的小分子NSC-741909可特异性杀伤吉非替尼原发耐药细胞,这是通过持续激活JNK途径从而导致细胞凋亡实现的.

Abstract：

Objective To study whether a recently identified novel anticancer agent NSC-741909 can suppresses the growth of non-small cell lung cancer cell line ( NSCLC) which has primary resistance to Gefitinib and explore its molecular mechanisms. Methods Select NSCLC cell line which is resistant to Gefitinib. Observe the cell growth supression effect of NSC-741909 to the cell line, apoptosis and actin cytoskeleton changement. Observe K-ras, JNK, p-JNK protein expression by Western blotting. Results NSC-741909 can induce apoptosis of Gefitinib resistant cell lines at 24 h. At that time point, FITC-A +/ PE-A + increased to (6. 9 ±0.6)% and (21. 1 ±3.2)% (P ＜0. 01) at 1 and 2 μmol/L;K-ras protein decreased to 70% at 2 h; p-JNK expression was increased and lasted for at least 15 h and total JNK remained the same. Cell cytoskeleton F-actin presented as loose, irregular and radiation arrangement. Conclusion NSC-741909 which supress the mutant K-ras expression can induce the apoptosis of the NSCLC which is primary resistant to Gefitinib. This inhibition was mediated by sustained JNK activation.

Effect of small molecule k-ras mutations of non-small cell lung cancer on gefitinib resistant cells

Objective To study whether a recently identified novel anticancer agent NSC-741909 can suppresses the growth of non-small cell lung cancer cell line ( NSCLC) which has primary resistance to Gefitinib and explore its molecular mechanisms. Methods Select NSCLC cell line which is resistant to Gefitinib. Observe the cell growth supression effect of NSC-741909 to the cell line, apoptosis and actin cytoskeleton changement. Observe K-ras, JNK, p-JNK protein expression by Western blotting. Results NSC-741909 can induce apoptosis of Gefitinib resistant cell lines at 24 h. At that time point, FITC-A +/ PE-A + increased to (6. 9 ±0.6)% and (21. 1 ±3.2)% (P ＜0. 01) at 1 and 2 μmol/L;K-ras protein decreased to 70% at 2 h; p-JNK expression was increased and lasted for at least 15 h and total JNK remained the same. Cell cytoskeleton F-actin presented as loose, irregular and radiation arrangement. Conclusion NSC-741909 which supress the mutant K-ras expression can induce the apoptosis of the NSCLC which is primary resistant to Gefitinib. This inhibition was mediated by sustained JNK activation.