达沙替尼体外抗鼻咽癌细胞增殖与转移的作用

Effects of SRC inhibitor dasatinib on nasopharyngeal carcinoma cells proliferation and invasion in vitro

目的 SRC作为酪氨酸蛋白激酶在多种实体肿瘤中存在高表达,通过激活RAS/RAF/MEK/ERK、PI3K/AKT通路促进肿瘤细胞的生长与增殖,并通过激活FAK促进肿瘤细胞的转移。此外,SRC还参与了EGFR抑制剂耐药的发生。本研究旨在研究SRC的抑制剂达沙替尼在体外对鼻咽癌细胞的抑制作用。方法采用MTT方法绘制细胞生长曲线,并计算达沙替尼对鼻咽癌细胞的半数抑制浓度(IC50值);PI/AnnexinV双染法检测达沙替尼诱导鼻咽癌细胞凋亡的作用;Western Blot检测达沙替尼引起鼻咽癌细胞中信号通路的改变;Transwell实验检测达沙替尼对鼻咽癌细胞迁移的影响。结果达沙替尼能明显在体外抑制鼻咽癌细胞的增殖,诱导鼻咽癌细胞的凋亡,并且呈浓度依赖性;应用达沙替尼处理CNE2后发现,能明显抑制RAS/RAF/MEK/ERK、PI3K/AKT通路活性表现为phospho-AKT、Phospho-MEK、Phospho-ERK的表达水平明显减低;达沙替尼还能明显抑制CNE2的迁移能力和Phospho-FAK的表达水平。结论 SRC的抑制剂达沙替尼能在体外明显抑制鼻咽癌细胞中RAS/RAF/MEK/ERK、PI3K/AKT通路的活性和FAK蛋白的表达,进一步抑制鼻咽癌细胞的增殖与转移,促进细胞凋亡。

Objective SRC tyrosine kinase is overexpressed in variety of solid tumors,which promotes cell growth and proliferation by ac tivating the RAS/RAF/MEK/ERK, PI3K/AKT pathway and promotes metastasis by improving FAK activity. In addition, SRC is also in volved in the resistance of EGFR inhibitors. This research aims to research the effects of dasatinib on nasopharyngeal carcinoma cells in vitro. Methods Cell growth＇ rate and 50% inhibitory concentration were calculated by MTI＂ assay, Dasatinibinduced apoptotic cells were investigated by Annexin V/PI staining. Protein expression from cell extracts was analyzed by Western Blot. Cell mobility was investigated by Transwell. Results Dasatinib significantly inhibited CNE2 proliferation and induced apoptosis in vitro;phospho-AKT, Phospho-MEK, Phospho-ERK expression were significantly reduced when treated with dasatinib which means the deregulated RAS/RAF/MEK/ERK, PI3K/AKT pathway activity;dasatinib CNE2 also significantly inhibited the mobility as well as Phospho-FAK expression. Conclusions Sre inhibitor dasatinib inhibited in vitro NPC ceils RAS/RAF/MEK/ERK, PI3K/AKT pathway activity and FAK protein expression, fur ther inhibiting the proliferation and metastasis of nasopharyngeal carcinoma cells and promote apoptosis.