Albayrak Gulsah, Korkmaz Funda Demirtas, Tozcu Duygu, Dogan Turacli Irem
Department of Medical Biology, Faculty of Medicine, Ufuk University, Ankara, Turkey.
Department of Medical Biology and Genetics, Faculty of Medicine, Gazi University, Ankara, Turkey.
J Cell Biochem. 2019 Jun;120(6):10564-10571. doi: 10.1002/jcb.28342. Epub 2019 Jan 10.
Lung cancer stands out as the most common cancer type worldwide. The most common genetic alteration detected in adenocarcinoma patients is KRAS. KRAS mutated patients still cannot get benefit from precision medicine approaches and lack a targeted therapy. Elesclomol is an investigational agent for melanoma and other malignancies. In this study, we evaluated its effect on cellular apoptosis, survival, and metastasis mechanisms on KRAS mutant A549 and Calu-1 cell lines.
The cytotoxic effects of Elesclomol on A549 and Calu-1 cells were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell viability test. Cells were treated with IC concentration and then apoptosis-related (Casp-3, Casp-9, Bcl-2, and Bcl-xL), survival-related (Akt, p-Akt, Erk, and p-Erk), and metastasis-related (E-cadherin, Vimentin, MMP-2, and MMP-9) protein expressions were determined by Western blot analysis. Elesclomol's effect on cell migration was evaluated by wound healing. Total oxidant, malondialdehyde (MDA), and glutathione (GSH) levels after Elesclomol treatment were assessed.
Elesclomol not only induced apoptotic proteins but also inhibited metastatic protein expressions and migration in both cells. Also, p-Erk activity was diminished by Elesclomol treatment as a reflection of decreased proliferation. However, p-Akt was enhanced as a cellular survival mechanism. Although Elesclomol's effects on oxidative stress parameters were puzzling, it induced total oxidant status (TOS), and MDA in Calu-1 cells.
Elesclomol might provide an alternative treatment approach for patients with KRAS mutant lung adenocarcinoma and other solid tumor malignancies that harbor KRAS mutations. This would enable the development of biomarker-driven targeted therapy for KRAS mutant adenocarcinoma patients.
肺癌是全球最常见的癌症类型。在腺癌患者中检测到的最常见基因改变是KRAS。KRAS突变患者仍无法从精准医学方法中获益，且缺乏靶向治疗。艾乐替尼是一种用于黑色素瘤和其他恶性肿瘤的研究药物。在本研究中，我们评估了其对KRAS突变的A549和Calu-1细胞系的细胞凋亡、存活及转移机制的影响。
通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）细胞活力试验测定艾乐替尼对A549和Calu-1细胞的细胞毒性作用。用半数抑制浓度（IC）处理细胞，然后通过蛋白质免疫印迹分析测定凋亡相关蛋白（Casp-3、Casp-9、Bcl-2和Bcl-xL）、存活相关蛋白（Akt、p-Akt、Erk和p-Erk）以及转移相关蛋白（E-钙黏蛋白、波形蛋白、基质金属蛋白酶-2和基质金属蛋白酶-9）的表达。通过伤口愈合实验评估艾乐替尼对细胞迁移的影响。评估艾乐替尼处理后的总氧化剂、丙二醛（MDA）和谷胱甘肽（GSH）水平。
艾乐替尼不仅诱导凋亡蛋白表达，还抑制两种细胞中的转移蛋白表达和迁移。此外，艾乐替尼处理降低了p-Erk活性，反映出增殖减少。然而，作为一种细胞存活机制p-Akt活性增强。尽管艾乐替尼对氧化应激参数的影响令人困惑，但它在Calu-1细胞中诱导了总氧化剂状态（TOS）和MDA。
艾乐替尼可能为KRAS突变的肺腺癌患者及其他携带KRAS突变的实体瘤恶性肿瘤患者提供一种替代治疗方法。这将有助于为KRAS突变的腺癌患者开发生物标志物驱动的靶向治疗。
