Effect of siRNAs targeting the EGFR T790M mutation in a non-small cell lung cancer cell line resistant to EGFR tyrosine kinase inhibitors and combination with various agents
Effect of siRNAs targeting the EGFR T790M mutation in a non-small cell lung cancer cell line resistant to EGFR tyrosine kinase inhibitors and combination with various agents
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DOI：
10.1016/j.bbrc.2012.12.070
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发表时间：
2013-02-15
影响因子：
3.1
通讯作者：
De Greve, Jacques
中科院分区：
文献类型：
--
作者：
Chen, Gang;Kronenberger, Peter;De Greve, Jacques
表皮生长因子受体（EGFR）是治疗非小细胞肺癌（NSCLC）的有效靶点。然而，一些突变赋予对当前可用药物的抗性，特别是经常发生的T790M突变。在目前的研究中，我们在包含L858R和T790M突变的非小细胞肺癌细胞系H1975中检测了T790M特异性sirna与其他egfr特异性sirna的作用。在H1975细胞中，T790M特异性sirna能够抑制T790M和EGFR mRNA，降低EGFR蛋白表达，抑制细胞生长，诱导细胞caspase活性。然而，与其他egfr特异性sirna相比，这种效应的效力较弱。egfr特异性sirna在H358、H1650、H292、HCC827和H1975细胞中强烈抑制细胞生长并诱导细胞凋亡，这些细胞对酪氨酸激酶抑制剂（TKIs）或西妥昔单抗的反应较弱。T790M特异性sirna的加入可以恢复T790M突变体H1975细胞对TKIs的敏感性。t790m特异性sirna与西妥昔单抗联合使用也增强了H1975细胞生长抑制和诱导凋亡的作用。在所测试的抗egfr药物中，当阿法替尼与t790m特异性sirna联合使用时，观察到最强的生物学效应。在H1975细胞中，阿法替尼与西妥昔单抗联合使用也提供了额外的效果。综上所述，sirna敲低T790M转录物进一步降低T790M突变肺癌细胞在TKIs或西妥昔单抗治疗下的细胞生长。有效的、不可逆的激酶抑制剂如阿法替尼与T790M特异性sirna联合使用，作为治疗含有耐药T790M突变的肺癌的新策略，应进一步研究。(C) 2012爱思唯尔公司版权所有。
The epidermal growth factor receptor (EGFR) is a validated therapeutic target in non-small cell lung cancer (NSCLC). However, some mutations confer resistance to current available agents, especially the frequently occurring T790M mutation. In the current study, we have examined, in a NSCLC cell line H1975 containing both L858R and T790M mutations, the effect of T790M-specific-siRNAs versus other EGFR-specific-siRNAs. T790M-specific-siRNAs were able to inhibit T790M and EGFR mRNA, to reduce EGFR protein expression, as well as to reduce the cell growth and induce cell caspase activity in H1975 cells. However, this effect showed less potency compared to the other EGFR-specific-siRNAs. EGFR-specific-siRNAs strongly inhibited cell growth and induced apoptosis in H358, H1650, H292, HCC827 and also in H1975 cells, which showed weak response to tyrosine kinase inhibitors (TKIs) or cetuximab. The addition of T790M-specific-siRNAs could rescue the sensitivity of T790M mutant H1975 cells to TKIs. The combination of T790M-specific-siRNAs and cetuximab also additively enhanced cell growth inhibition and induction of apoptosis in H1975 cells. Among the anti-EGFR agents tested, the strongest biological effect was observed when afatinib was combined with T790M-specific-siRNAs. Afatinib also offered extra effect when combined with cetuximab in H1975 cells. In conclusion, knock-down of T790M transcript by siRNAs further decreases the cell growth of T790M mutant lung cancer cells that are treated with TKIs or cetuximab. The combination of a potent, irreversible kinase inhibitor such as afatinib, with T790M-specific-siRNAs should be further investigated as a new strategy in the treatment of lung cancer containing the resistant T790M mutation. (C) 2012 Elsevier Inc. All rights reserved.