塞来昔布诱导人肝癌细胞株QGY-7701凋亡的研究

目的探讨塞来昔布体外诱导人肝癌细胞株QGY-7701凋亡的作用。方法设立空白对照组(培养液中不加任何药物)及塞来昔布5、25、50、100、150μmol.L-1 5个剂量组,作用时间分为24、48、72 h,应用四甲基偶氮唑盐(MTT)法观察塞来昔布不同浓度和作用时间对QGY-7701细胞生存率的影响,选择合适的作用时间和3个剂量再做其他凋亡方法的检测;采用Hoechst 33258染色荧光显微镜观察凋亡细胞的形态学变化;采用双荧光染色(AnnexinV-EGFP/PI)流式细胞检测、末端脱氧核苷酰基转移酶介导性dUTP切口末端标记(TUNEL)检测、DNA含量分析(检测细胞周期)等方法多指标检测细胞凋亡率。结果通过Hoechst 33258染色可以从细胞形态上看出塞来昔布诱导QGY-7701细胞凋亡,而Annexin V-EGFP/PI流式细胞检测、TUNEL检测、细胞周期及MTT检测结果显示随着塞来昔布浓度升高和作用时间延长,凋亡率升高(P<0.05,P<0.01)。结论塞来昔布有诱导肝癌细胞QGY-7701凋亡,抑制其生长的作用,具有治疗肝癌的潜在功能。

Apoptosis of human hepatoma carcinoma cell strain QGY-7701 induced by celecoxib

Objective To investigate the function of celecoxib in inducing apoptosis of human liver cancer cell line QGY-7701.Methods One control group（medium without any drug）and five dosage groups with celecoxib 5,25,50,100,150 μmol·L-1 were established with action time 24,48,72 hours.The cytostatic effect of celecoxib on QGY-7701 cell was observed by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT）.Three doses were detected by other apoptotic method at appropriate action time.After staining by Hoechst 33258,apoptotic cell morphology change was assessed by fluoroscope.The cell apoptosis rate was assessed by double fluorescent staining（Annexin V-EGFP/PI） and flow cytometry（FCM）,terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling（TUNEL） detection and cell cycle analysis.Results Observing from cell morphous,apoptosis of cells QGY-7701 which induced by celecoxib could be found by Hoechst 33258 dyeing.The findings of Annexin V-EGFP/PI,TUNEL cell cycle assay and MTT showed that the apoptosis rate increased with the increasing concentration and action time of celecoxib.Conclusion Celecoxib can inhibit the proliferation of QGY-7701 by inducing its apoptosis,which suggests celecoxib has potential function in curing liver cancer.