DNA Polymerase Alpha Subunit B Is a Binding Protein for Erlotinib Resistance in Non-Small Cell Lung Cancer.
DNA Polymerase Alpha Subunit B Is a Binding Protein for Erlotinib Resistance in Non-Small Cell Lung Cancer.
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DOI：
10.3390/cancers12092613
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发表时间：
2020-09-13
期刊：
影响因子：
5.2
通讯作者：
Kwon HJ
中科院分区：
文献类型：
--
作者：
Kim TY;Ji ES;Lee JY;Kim JY;Yoo JS;Szasz AM;Dome B;Marko-Varga G;Kwon HJ
非小细胞肺癌（NSCLC）占所有肺癌的近85%，是整体癌症死亡率的主要贡献者。厄洛替尼通过抑制上皮生长因子受体（EGFR）激酶活性来治疗NSCLC。尽管它的疗效很高，但对药物产生耐药性的患者可能会复发。我们进行DARTS LC-MS/MS与SWATH的DIA分析，并确定了一种新的结合蛋白的厄洛替尼，可能是NSCLC耐药的基础。我们的研究表明，厄洛替尼结合POLA 2除了EGFR。DARTS和CETSA结果证实了这一点。重要的是，四种NSCLC细胞系中的POLA 2表达水平与抗增殖厄洛替尼功效正相关（Pearson相关系数，R = 0.9886）。这些结果表明，POLA 2是一种新的互补靶蛋白的厄洛替尼，并可能在临床上提供有效性作为一个替代标记物的耐药与NSCLC患者。厄洛替尼抑制上皮生长因子受体（EGFR）激酶活性，用于治疗非小细胞肺癌（NSCLC）。尽管它的疗效很高，但对药物产生耐药性的患者可能会复发。为了解决厄洛替尼耐药的潜在机制，我们研究了与药物作用模式相关的其他机制，通过多种蛋白质结合相互作用，除了EGFR，通过使用药物亲和力响应靶稳定性（DARTS）和液相色谱-质谱（LC-MS/MS）方法与非标记的厄洛替尼。DNA聚合酶α亚基B（POLA 2）被鉴定为一种新的厄洛替尼结合蛋白，通过DARTS平台进行了验证，并辅以细胞热位移试验。POLA 2的基因敲低促进了药物在具有高POLA 2表达的厄洛替尼耐药细胞系H1299中的抗增殖作用，而POLA 2的过表达恢复了具有低POLA 2表达的厄洛替尼敏感细胞系HCC 827中的抗增殖作用。重要的是，四种NSCLC细胞系中的POLA 2表达水平与抗增殖厄洛替尼功效正相关（Pearson相关系数，R = 0.9886）。这些结果表明，POLA 2是一种新的互补靶蛋白的厄洛替尼，并可能在临床上提供有效性作为一个替代标记物的耐药与NSCLC患者。
Non-small-cell lung carcinoma (NSCLC) covers for almost 85% of all lung cancers and a major contributor to the overall cancer death rate. Erlotinib is used to treat NSCLC via inhibition of epithelial growth factor receptor (EGFR) kinase activity. Despite its high efficacy, recurrence can occur in patients who become resistant to the drug. We performed DARTS LC-MS/MS with SWATH of DIA analysis and identified a novel binding protein of Erlotinib that may underlie NSCLC resistance. Our study indicated that Erlotinib binds POLA2 in addition to EGFR. This was confirmed by DARTS and CETSA results. Importantly, POLA2 expression levels in four NSCLC cell lines were positively correlated with anti-proliferative Erlotinib efficacy (Pearson correlation coefficient, R = 0.9886). These results suggest that POLA2 is a novel complementary target protein of Erlotinib, and could clinically provide validity as a surrogate marker for drug resistance in patients with NSCLC. Erlotinib inhibits epithelial growth factor receptor (EGFR) kinase activity and is used to treat non-small cell lung cancer (NSCLC). Despite its high efficacy, recurrence can occur in patients who become resistant to the drug. To address the underlying mechanism of Erlotinib resistance, we investigated additional mechanisms related to mode-of-drug-action, by multiple protein-binding interactions, besides EGFR by using drug affinity responsive target stability (DARTS) and liquid chromatography-mass spectrometry (LC-MS/MS) methods with non-labeled Erlotinib. DNA polymerase alpha subunit B (POLA2) was identified as a new Erlotinib binding protein that was validated by the DARTS platform, complemented with cellular thermal shift assays. Genetic knock-down of POLA2 promoted the anti-proliferative effect of the drug in the Erlotinib-resistant cell line H1299 with high POLA2 expression, whereas the overexpression of POLA2 restored anti-proliferative effects in the Erlotinib-sensitive cell line HCC827 with low POLA2 expression. Importantly, POLA2 expression levels in four NSCLC cell lines were positively correlated with anti-proliferative Erlotinib efficacy (Pearson correlation coefficient, R = 0.9886). These results suggest that POLA2 is a novel complementary target protein of Erlotinib, and could clinically provide validity as a surrogate marker for drug resistance in patients with NSCLC.
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影响因子：
5.3
作者：
Neumair, P.;Joos, L.;Joerger, M.
通讯作者：
Joerger, M.
影响因子：
4.2
作者：
Qiao, Haishi;Zhao, Dan;Li, Erguang
通讯作者：
Li, Erguang
通过抑制磷酸甘油酸脱氢酶克服 EGFR 突变阳性肺腺癌的厄洛替尼耐药性
影响因子：
12.4
作者：
Dong JK;Lei HM;Liang Q;Tang YB;Zhou Y;Wang Y;Zhang S;Li WB;Tong Y;Zhuang G;Zhang L;Chen HZ;Zhu L;Shen Y
通讯作者：
Shen Y
DOI：
10.6004/jnccn.2019.0059
发表时间：
2019-12-01
影响因子：
13.4
作者：
Ettinger, David S.;Wood, Douglas E.;Hughes, Miranda
通讯作者：
Hughes, Miranda
DOI：
10.1073/pnas.0710370104
发表时间：
2007-12-26
影响因子：
11.1
作者：
Bean, James;Brennan, Cameron;Pao, William
通讯作者：
Pao, William