5-氮-2'-脱氧胞苷诱导膀胱癌BIU87细胞凋亡的研究

目的:研究5-氮-2'-脱氧胞苷(5-Aza-2'-decxycytidine,5-Aza-CdR)对膀胱癌BIU87细胞凋亡的影响。方法:以不同浓度的5-氮-2'-脱氧胞苷作用于膀胱癌BIU87细胞,以四甲基偶氮唑蓝(MTT)法检测细胞的增殖程度,AnnexinV-FITC/PI双染流式细胞仪(flow cytometry,FCM)检测凋亡率,AO荧光染色对细胞凋亡形态学观察,同时应用Western blot检测膀胱癌BIU87细胞RASSF1A蛋白表达的改变。结果:BIU87细胞加入(0.1、0.5、1.0、5.0)nmol/L的5-Aza-CdR后与对照组相比各组BIU87细胞增长速度减慢。不同浓度的5-Aza-CdR可以显著降低BIU87细胞的存活率,与对照相组比差异显著(P<0.05)。BIU87细胞的凋亡率逐渐增高且与5-Aza-CdR呈浓度依赖关系(P<0.05)。荧光显微镜下见典型的凋亡形态学改变,随5-Aza-CdR浓度的升高BIU87细胞RASSF1A蛋白的表达也升高。结论:5-Aza-CdR可能通过对RASSF1A基因的再表达而抑制肿瘤细胞的生长,从而诱导膀胱癌BIU87细胞凋亡的增加。

Study on apoptosis of BIU87 cell line induced by 5-Aza-CdR

Objective: To investigate the apoptosis of bladder cancer cell line-BIU87 cell by 5-Aza-CdR.Methods: Cultured BIU87 cells were treated with variable doses of 5-Aza-CdR. MTT was used to detect the proliferation of BIU87 cell;The AnnexinV-FITC/PI double staining analysis by flow cytometry was used to evaluate apoptosis rate.AO fluorescent staining was used to observe apoptosis structure feature of cells.The changes of RASSF1A protein expression were detected by western blot.Results: BIU87 cells were treated with different doses of 5-Aza-CdR[(0.1,0.5,1.0,5.0)nmol/ L],displayed a slowly growth rate in contrast to the control group.Different doses of 5-Aza-CdR could obviously decrease the viability rate and obviously increase the apoptotic rate of BIU87 cells respectively compared with control group(P<0.05),the apoptotic rate was dose-dependant(P<0.05).The treated BIU87 cells exhibited some morphological features of apoptosis under the fluorescence microscope.The expression of RASSF1A protein increased with the doses of 5-Aza-CdR.Conclusion: 5-Aza-CdR may inhibit growth by reacting RASSF1A gene and induce BIU87 cells apoptosis;it has an inhibiting effect on proliferation of BIU87 cells.