小分子酪氨酸激酶抑制剂安罗替尼对鼻咽癌细胞的抗肿瘤活性及其机制

目的 探索小分子酪氨酸激酶抑制剂安罗替尼对鼻咽癌细胞的抗肿瘤作用及其潜在机制，为临床应用安罗替尼治疗鼻咽癌提供可靠的实验证据。方法 通过CCK-8实验和克隆形成实验检测不同剂量梯度安罗替尼对鼻咽癌细胞6-10B和SUNE-1的细胞活性、增殖能力的影响；通过Transwell和划痕实验分析安罗替尼对鼻咽癌细胞侵袭及迁移能力的影响；利用流式细胞术检测安罗替尼对鼻咽癌细胞周期及凋亡的影响；采用Western blotting验证安罗替尼处理后鼻咽癌细胞凋亡相关蛋白的表达变化。结果 在鼻咽癌细胞中加入安罗替尼处理后，6-10B实验组和SUNE-1实验组细胞存活分数降低，细胞集落形成数量明显减少，且呈显著的剂量依赖性，在药物浓度为2.0μmol·L^-1时可达到约50%的增殖及克隆抑制率(P<0.01)。Western blotting分析发现，安罗替尼可能是由Erk通路介导线粒体通路激活，促进鼻咽癌细胞凋亡，且以药物浓度达到2.0μmol·L^-1时作用最为明显。凋亡检测结果证实，2.0μmol·L^-1安罗替尼处理后的实验组较...

Anti-tumor activity and mechanism of the small molecular tyrosine kinase inhibitor anlotinib in nasopharyngeal cancer cells

Objective To explore the anti-tumor effects of anlotinib,a small molecular tyrosine kinase inhibitor,on nasopharyngeal carcinoma (NPC) cells and its potential mechanism,and to provide reliable experimental evidence for the clinical treatment of NPC with anlotinib.Methods The responses of serial dose gradients of anlotinib on the cell viability and proliferation of 6-10B and SUNE-1 cell lines were detected by cell proliferation viability assay and colony formation assay.Then,the downstream pathway proteins in cell lines after anlotinib treatment were visualized by Western blotting.The effects of promoting apoptosis and inducing cell cycle arrest by anlotinib were confirmed through flow cytometry.The capacity of anlotinib in inhibiting migration and invasion of 6-10B and SUNE-1 cell lines was verified by transwell assay and wound healing assay.Results After treatment of anlotinib,cell viability and proliferation were inhibited in nasopharyngeal carcinoma 6-10B and SUNE-1 cell lines in a dose-dependent way,and the inhibition rate was up to 50%when then concentration of anlotinib was 2μmol·L^-1(P<0.01).Western blotting results showed that anlotinib might induce the cell apoptosis by activating the mitochondrial pathway and potentially the Erk pathway,with the highest apoptosis rate when the anlotinib concentration up to 2μmol·L^-1.The results of flow cytometry showed that 2μmol·L^-1anlotinib induced remarkable cell apoptosis,with early apoptosis predominantly (P<0.001).What’s more,the nasopharyngeal carcinoma cells had obvious changes in cycle distribution after treated with 2μmol·L^-1anlotinib.Both 6-10B and SUNE-1 cells had a G₂/M arrest,and there was a significant difference in 6-10B group (P=0.0004),but no statistical differences in SUNE-1 group (P=0.0723).By transwell assay and wound healing assay,the significant suppression of invasion and migration in 6-10B and SUNE-1 was definitude (P<0.05).Conclusion This study indicated that anlotinib promoted the apoptosis and inhibited the proliferation of nasopharyngeal carcinoma cells 6-10B and SUNE-1 through the Erk pathway.It could be a potential target drug for nasopharyngeal carcinoma.