The afatinib resistance of in vivo generated H1975 lung cancer cell clones is mediated by SRC/ERBB3/c-KIT/c-MET compensatory survival signaling.
The afatinib resistance of in vivo generated H1975 lung cancer cell clones is mediated by SRC/ERBB3/c-KIT/c-MET compensatory survival signaling.
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DOI：
10.18632/oncotarget.7746
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发表时间：
2016-04-12
期刊：
影响因子：
--
通讯作者：
Dent P
中科院分区：
文献类型：
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作者：
Booth L;Roberts JL;Tavallai M;Webb T;Leon D;Chen J;McGuire WP;Poklepovic A;Dent P
我们通过将已建立的肿瘤短暂暴露于药物，并收集再生长的肿瘤，产生H1975肺癌细胞的阿法替尼耐药克隆。与对照克隆相比，阿法替尼耐药H1975克隆在原癌基因中未显示任何额外突变。阿法替尼耐药H1975肿瘤克隆表达的PTEN低于对照克隆，在阿法替尼耐药克隆中，这与基础SRC Y 416、ERBB 3 Y1289、AKT T308和mTOR S2448磷酸化增加、ERBB 1、ERBB 2和ERBB 3表达降低以及c-MET、c-KIT和PDGFRβ总表达增加相关。阿法替尼耐药克隆通过[ERBB 3 + c-MET + c-KIT]敲低而非单个或双联体敲低组合选择性杀死。ERBB 1/2/4抑制剂阿法替尼与SRC家族抑制剂达沙替尼联合使用，以大于相加的方式杀死阿法替尼耐药H1975细胞;与达沙替尼联合使用的其他药物（如舒尼替尼、克唑替尼和amufatinib）的有效性较低。[阿法替尼+达沙替尼]处理深度灭活了H1975阿法替尼耐药克隆中的ERBB 3、AKT和mTOR，并增加了ATG 13 S318磷酸化。ATG 13、Beclin 1或eIF 2 α的敲低强烈抑制了[ERBB 3 + c-MET + c-KIT]敲低的杀伤作用，但仅对[阿法替尼+达沙替尼]致死性具有中度保护作用。因此，阿法替尼耐药H1975 NSCLC细胞依赖于残留ERBB 3的ERBB 1和SRC依赖性超活化以及野生型c-MET和c-KIT蛋白表达升高导致的信号传导升高来保持存活。通过SRC和ERBB 1的阻断来抑制ERBB 3信号传导导致肿瘤细胞死亡。
We generated afatinib resistant clones of H1975 lung cancer cells by transient exposure of established tumors to the drug and collected the re-grown tumors. Afatinib resistant H1975 clones did not exhibit any additional mutations in proto-oncogenes when compared to control clones. Afatinib resistant H1975 tumor clones expressed less PTEN than control clones and in afatinib resistant clones this correlated with increased basal SRC Y416, ERBB3 Y1289, AKT T308 and mTOR S2448 phosphorylation, decreased expression of ERBB1, ERBB2 and ERBB3 and increased total expression of c-MET, c-KIT and PDGFRβ. Afatinib resistant clones were selectively killed by knock down of [ERBB3 + c-MET + c-KIT] but not by the individual or doublet knock down combinations. The combination of the ERBB1/2/4 inhibitor afatinib with the SRC family inhibitor dasatinib killed afatinib resistant H1975 cells in a greater than additive fashion; other drugs used in combination with dasatinib such as sunitinib, crizotinib and amufatinib were less effective. [Afatinib + dasatinib] treatment profoundly inactivated ERBB3, AKT and mTOR in the H1975 afatinib resistant clones and increased ATG13 S318 phosphorylation. Knock down of ATG13, Beclin1 or eIF2α strong suppressed killing by [ERBB3 + c-MET + c-KIT] knock down, but were only modestly protective against [afatinib + dasatinib] lethality. Thus afatinib resistant H1975 NSCLC cells rely on ERBB1- and SRC-dependent hyper-activation of residual ERBB3 and elevated signaling, due to elevated protein expression, from wild type c-MET and c-KIT to remain alive. Inhibition of ERBB3 signaling via both blockade of SRC and ERBB1 results in tumor cell death.