藤黄酸衍生物-5诱导人头颈部鳞状细胞癌细胞凋亡的体外研究

Gambogic acid derivative compound 5 induces apoptosis in human head and neck squamous cell carcinoma in vitro

目的:研究藤黄酸(GA)的衍生物5(C5),体外诱导人头颈部鳞状细胞癌(HNSCC)细胞的凋亡作用。方法:不同时间段分别用不同剂量C5作用于HNSCC细胞系CAL27、HN13和HN30,采用MTT法检测细胞存活率,Logit法测定抑制50%的细胞生长所需的浓度(IC50)值。采用Annexin-V/PI双染色流式细胞仪检测细胞凋亡。免疫细胞化学法和Western blot法检测凋亡相关蛋白表达的变化。结果:C5明显抑制CAL27、HN13和HN30细胞的生长,且与剂量和时间呈正相关。IC50值比GA对照组明显降低。应用5.0μmol/L高浓度的C5,处理CAL27细胞系组24 h、48 h和72 h,细胞凋亡比例分别为52.19%、65.24%和84.53%。C5引起明显的、剂量依赖性的Bcl-2表达减少和Bax表达增加。结论:C5可以体外通过上调Bax蛋白和下调Bcl-2蛋白表达诱导头颈部鳞状细胞癌细胞的凋亡。

Objective： To observe the effect of gambogic acid（GA） derivative compound 5（C5） on apoptosis in human head and neck squamous cell carcinoma（HNSCC） in vitro. Methods： In the present study,HNSCC cell lines CAL27,HN13 and HN30 were treated with C5 at different doses for different times. The cell viability was detected with MTT assay and the IC50 values were also determined by Logit method. Cell apoptotic rates induced by C5 in the three cell lines were observed by Annexin-V / PI doubling staining by the flow cytometry assay. The changes on the expressions of apoptosis-related proteins were detected by Western blot and fluorescent immunocytochemistry assay. Results： CAL27,HN13 and HN30 cell lines were dramatically inhibited in a dose- and time-dependent manner after incubation with C5. The IC50 values were less than GA. When CAL27 cells were incubated with high dose C5 of 5.0μmol /L for 24,48 and 72 h,the apoptotic rates were about 52.19%,65.24% and 84.53%,respectively. C5 decreased protein expression levels of Bcl-2 and increased protein levels of Bax in the CAL27,HN13 and HN30 cell lines. Conclusions： Altogether,our data demonstrated that C5 induced apoptosis of HNSCC cell lines with up-regulation of Bax and down-regulation of Bcl-2 in vitro.