摘要
目的探讨5-Aza-2′-dC增加肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导乳腺癌细胞凋亡敏感性的作用及其可能机制。方法 5μmol/L 5-Aza-2′-dC预处理细胞96 h,TRAIL(0,50,100,200 ng/mL)处理细胞12 h,MTT检测细胞增殖抑制率,Annexin V-FITC/PI法检测细胞凋亡,流式细胞技术检测TRAIL受体DR4及DR5的表达,Western blot检测caspase-8蛋白表达水平,Real-time PCR检测细胞DR4、DR5和caspase-8 mRNA表达。结果 5-Aza-2′-dC预处理组与对照组相比,TRAIL对细胞增殖抑制率和诱导凋亡率明显高于对照组(P<0.05)。5-Aza-2′-dC能增加细胞表面DR4和DR5表达,促进凋亡通路关键蛋白caspase-8活化。5-Aza-2′-dC在mRNA水平增加了DR4、DR5和caspase-8的表达。结论 5-Aza-2′-dC增加了TRAIL诱导乳腺癌细胞凋亡的敏感性,其可能机制是通过上调DR4、DR5和caspase-8表达实现。
Objective To observe the effect of 5-Aza-2＇-dC on TRAIL-induced apoptosis of breast cancer cells and explore the potential mechanism. Methods Human breast caneer cells MDA-MB-231 were pretreated tbr 96 h with/without 5-Aza-2＇-dC （5 txmol/L）,and se- quentially treated with TRAIL（0,50,100,200 ng/mL ） for 12 h. MTT assay was used to deteet the cell proliferation and Annexin V-FITC/PI was apphed to analyze eell apoptosis. The expression of DR4 and DR5 was analyzed by flow-cytometry and the expression of caspase-8 was determined by Western blot. The mRNA expression of DRd, DR5 and caspase-8 was analyzed by Real-time PCR. Results The sequential treatment with 5-Aza-2＇-dC and TRAIL resuhed in significant inhibition of the cell proliferation and apoptosis. Moreover,pretreatment of breast caneer cells with 5-Aza-2＇-dC caused enhancement of the expression of DR4, DR5 and caspase-8 both in protein and mRNA levds. Conclusion These findings suggest that pretreatment with 5-Aza-2＇-dC enhances TRAIL-induced apoptosis in breast caneer cells. 5-Aza-2＇ -dC mediated up-regulation of DR4, DR5 and caspase-8 expression may be the potential mechanism.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2012年第12期1102-1105,共4页
Journal of China Medical University
基金
辽宁省教育厅高校科研计划(L2012281)
