Department of Biochemistry and Molecular Biology, Institute of Pathology and Pathophysiology, School of Basic Medical Science, China Medical University, Shenyang, Liaoning 110001, P.R. China.
Oncol Rep. 2013 Jul;30(1):139-48. doi: 10.3892/or.2013.2424. Epub 2013 Apr 24.
Lung cancer accounts for 13% (1.6 million) of the total cases and 18% (1.4 million) of the deaths in 2008. Crizotinib (PF-02341066) is identified as an ATP competitive small-molecular inhibitor for anaplastic lymphoma kinase (ALK). The US Food and Drug Administration (FDA) approved crizotinib to be used for the treatment of patients with locally advanced or metastatic ALK-positive NSCLC in 2011. In the present study, the side population (SP) and main population (MP) cells were obtained from Lewis lung carcinoma cells (LLC) and analyzed by DNA dye (Hoechst 33342) and flow cytometry. LLC SP and MP cells were confirmed as no ALK fusion gene by fluorescence in situ hybridization. The effects of crizotinib on LLC SP and MP cells both in vivo and in vitro were identified. Our results indicate that crizotinib can induce apoptosis and G1 phase arrest in LLC MP cells. Crizotinib used in combination with verapamil can inhibit proliferation of LLC SP cells. Moreover, crizotinib decreased tumor size and weight and inhibited angiogenesis in established xenografted tumors. To analyze the signaling pathway involved, computer simulation, Affymetrix microarray analysis and western blot analysis were performed. In these assays, crizotinib was found to dock into Smad3 and activate the Smad signaling pathway. Overall, these studies demonstrate the antitumor activity of crizotinib in LLC cell line, and provide a novel use for crizotinib.
肺癌占 2008 年总病例数的 13%（160 万）和总死亡数的 18%（140 万）。克唑替尼（PF-02341066）被鉴定为一种间变性淋巴瘤激酶（ALK）的 ATP 竞争性小分子抑制剂。美国食品和药物管理局（FDA）于 2011 年批准克唑替尼用于治疗局部晚期或转移性 ALK 阳性非小细胞肺癌患者。在本研究中，从 Lewis 肺癌细胞（LLC）中获得侧群（SP）和主群（MP）细胞，并通过 DNA 染料（Hoechst 33342）和流式细胞术进行分析。通过荧光原位杂交证实 LLC SP 和 MP 细胞无 ALK 融合基因。鉴定了克唑替尼对 LLC SP 和 MP 细胞的体内和体外作用。结果表明，克唑替尼可诱导 LLC MP 细胞凋亡和 G1 期阻滞。克唑替尼与维拉帕米联合使用可抑制 LLC SP 细胞的增殖。此外，克唑替尼可减小已建立的异种移植瘤的肿瘤大小和重量，并抑制血管生成。为了分析涉及的信号通路，进行了计算机模拟、Affymetrix 微阵列分析和 Western blot 分析。在这些测定中，发现克唑替尼与 Smad3 结合并激活 Smad 信号通路。总之，这些研究表明克唑替尼在 LLC 细胞系中具有抗肿瘤活性，并为克唑替尼提供了新的用途。
