Yoneyama Remi, Aoshiba Kazutetsu, Furukawa Kinya, Saito Makoto, Kataba Hiroaki, Nakamura Hiroyuki, Ikeda Norihiko
Department of Thoracic Surgery, Tokyo Medical University Hospital, Shinjuku-ku, Tokyo 160-0023, Japan; Department of Chest Surgery, Tokyo Medical University Ibaraki Medical Center, Inashiki-gun, Ibaraki 300-0395, Japan.
Department of Respiratory Medicine, Tokyo Medical University Ibaraki Medical Center, Inashiki-gun, Ibaraki 300-0395, Japan.
Oncol Lett. 2016 Jan;11(1):673-677. doi: 10.3892/ol.2015.3930. Epub 2015 Nov 17.
Cigarette smoking not only promotes lung carcinogenesis, but it has also been demonstrated to promote the progression of lung cancer. Despite nicotine being a major component of cigarette smoke, it is not carcinogenic when acting alone. Instead, it is believed to function as a tumor promoter. Due to the fatal consequences of lung cancer being primarily associated with the processes of invasion and metastasis, the present study aimed to determine the effect of nicotine on the migratory activity of lung cancer cells. The effect of nicotine on the migration of lung cancer A549 cells was evaluated by a wound healing assay. Hepatocyte growth factor (HGF) was used as a pro-migratory stimulus. During several of the experiments, specific inhibitors of α7-nicotine acetylcholine receptor (α7-nAchR), phosphoinositide kinase-3 (PI3K) and extracellular signal-related kinase (ERK)1/2 were included. The phosphorylation levels of Akt and ERK1/2 were examined using a cell-based protein phosphorylation assay. It was observed that nicotine did not induce cell migration by itself, but that it instead promoted HGF-induced cell migration. The effects of nicotine were inhibited by the pretreatment of the cells with the α7-nAchR inhibitor, methyllycaconitine, and the PI3K inhibitor, LY294002. The mitogen-activated protein kinase/ERK kinase kinase inhibitor exerted modest, but non-significant inhibitory activity on the effect of nicotine. Nicotine did not induce Akt phosphorylation by itself, but instead promoted the HGF-induced phosphorylation of Akt. It was also observed that nicotine had no effect on ERK1/2 phosphorylation. The results from the present study indicate that nicotine, when alone, does not have a pro-migratory function, but instead enhances responsiveness to the pro-migratory stimulus emitted by HGF. The current study provides an insight into the mechanism of tumor promotion by demonstrating that nicotine and α7-nAchRs act in synergy with the HGF-induced PI3K/Akt signaling pathway, increasing the sensitivity of lung cancer cells to HGF, and thereby promoting cell migration, a vital step in invasion and metastasis.
吸烟不仅会促进肺癌的发生,而且业已证明它还会促进肺癌的进展。尽管尼古丁是香烟烟雾的主要成分,但单独作用时它并无致癌性。相反,它被认为起到肿瘤促进剂的作用。由于肺癌的致命后果主要与侵袭和转移过程相关,本研究旨在确定尼古丁对肺癌细胞迁移活性的影响。通过伤口愈合试验评估尼古丁对肺癌A549细胞迁移的影响。肝细胞生长因子(HGF)用作促迁移刺激物。在若干实验中,纳入了α7-尼古丁乙酰胆碱受体(α7-nAchR)、磷酸肌醇激酶-3(PI3K)和细胞外信号调节激酶(ERK)1/2的特异性抑制剂。使用基于细胞的蛋白质磷酸化试验检测Akt和ERK1/2的磷酸化水平。观察到尼古丁本身不会诱导细胞迁移,而是促进HGF诱导的细胞迁移。用α7-nAchR抑制剂甲基lycaconitine和PI3K抑制剂LY294002对细胞进行预处理可抑制尼古丁的作用。促分裂原活化蛋白激酶/ERK激酶激酶抑制剂对尼古丁的作用有一定但不显著的抑制活性。尼古丁本身不会诱导Akt磷酸化,而是促进HGF诱导的Akt磷酸化。还观察到尼古丁对ERK1/2磷酸化没有影响。本研究结果表明,尼古丁单独作用时不具有促迁移功能,而是增强对HGF发出的促迁移刺激的反应性。当前研究通过证明尼古丁和α7-nAchRs与HGF诱导的PI3K/Akt信号通路协同作用,增加肺癌细胞对HGF的敏感性,从而促进细胞迁移(侵袭和转移的关键步骤),深入了解了肿瘤促进机制。