NSCLC Mutated Isoforms of CCDC6 Affect the Intracellular Distribution of the Wild Type Protein Promoting Cisplatinum Resistance and PARP Inhibitors Sensitivity in Lung Cancer Cells
NSCLC Mutated Isoforms of CCDC6 Affect the Intracellular Distribution of the Wild Type Protein Promoting Cisplatinum Resistance and PARP Inhibitors Sensitivity in Lung Cancer Cells
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DOI：
10.3390/cancers12010044
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发表时间：
2020-01-01
期刊：
影响因子：
5.2
通讯作者：
Celetti, Angela
中科院分区：
文献类型：
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作者：
Cerrato, Aniello;Morra, Francesco;Celetti, Angela
CCDC 6通过同源重组（HR）参与细胞周期检查点和DNA损伤修复。在NSCLC中，约30%的患者几乎不表达CCDC 6，约1%的患者检测到CCDC 6基因与RET和ROS激酶的重排。最近，已在原发性NSCLC中鉴定出命名为E227 K、S351 Y、N394 Y和T462 A的CCDC 6点突变。在这项工作中，我们分析了CCDC 6突变亚型对肺癌细胞的影响。通过下拉实验和免疫荧光，我们评估了CCDC 6肺突变体对CCDC 6野生型蛋白的生化和形态学影响。通过使用两种HR-报告基因测定，我们分析了CCDC 6肺突变体在HR过程中干扰CCDC 6生理学的作用。最后，通过细胞滴度测定，我们评估了表达CCDC 6突变体的肺癌细胞对不同药物治疗的反应。这项工作表明，迄今为止在NSCLC中鉴定的CCDC 6突变和截短亚型影响了野生型蛋白的细胞内分布，并损害了HR过程中的CCDC 6功能，最终诱导了肺癌细胞中的顺铂耐药性和PARP抑制剂敏感性。鉴定涉及CCDC 6基因产物的选定分子改变可能定义NSCLC个体化治疗的预测性生物标志物。
CCDC6 is implicated in cell cycle checkpoints and DNA damage repair by homologous recombination (HR). In NSCLC, CCDC6 is barely expressed in about 30% of patients and CCDC6 gene rearrangements with RET and ROS kinases are detected in about 1% of patients. Recently, CCDC6 point-mutations naming E227K, S351Y, N394Y, and T462A have been identified in primary NSCLC. In this work, we analyze the effects exerted by the CCDC6 mutated isoforms on lung cancer cells. By pull-down experiments and immunofluorescence, we evaluated the biochemical and morphological effects of CCDC6 lung-mutants on the CCDC6 wild type protein. By using two HR-reporter assays, we analyzed the effect of CCDC6 lung-mutants in perturbing CCDC6 physiology in the HR process. Finally, by cell-titer assay, we evaluated the response to the treatment with different drugs in lung cancer cells expressing CCDC6 mutants. This work shows that the CCDC6 mutated and truncated isoforms, identified so far in NSCLC, affected the intracellular distribution of the wild type protein and impaired the CCDC6 function in the HR process, ultimately inducing cisplatinum resistance and PARP-inhibitors sensitivity in lung cancer cells. The identification of selected molecular alterations involving CCDC6 gene product might define predictive biomarkers for personalized treatment in NSCLC.