摘要
目的观察5-杂氮-2′-脱氧胞苷(5-Aza-CdR)对顺铂耐药胃癌SGC-7901细胞14-3-3σ基因甲基化的影响;探讨14-3-3σ基因甲基化与顺铂耐药胃癌SGC-7901细胞耐药的关系。方法建立人胃癌顺铂耐药细胞株SGC-7901/CDDP,并用特异性甲基化抑制物5-Aza-CdR干预。MSP法检测14-3-3σ基因甲基化状态,蛋白质印迹法检测14-3-3σ蛋白表达,MTT法检测细胞增殖活性,流式细胞术分析细胞周期与凋亡。结果 SGC-7901/CDDP细胞的14-3-3σ基因高度甲基化并沉默,经过5、10μmol.L-15-Aza-CdR处理后,14-3-3σ蛋白重新表达,细胞发生顺铂耐药逆转,细胞活性抑制、在G0/G1期出现阻滞以及凋亡率明显增加。结论 5-Aza-CdR具有抑制顺铂耐药胃癌SGC-7901细胞14-3-3σ基因甲基化的作用,同时,顺铂作用与耐药机制部分依赖于14-3-3σ基因。
Aim To investigate the effects of 5-Aza-2′-deoxycytidine（5-Aza-CdR） on the methylation of 14-3-3σ in cisplatin-resistant gastric cancer line SGC-7901（SGC-7901/CDDP） and probe into the relationship between 14-3-3σ methylation and SGC-7901/CDDP cells.Methods SGC-7901/CDDP cells were treated with various concentrations of 5-Aza-CdR.The status of 5′CpG island methylation of 14-3-3σ gene in SGC-7901/CDDP cells was analyzed using methylation specific polymerase chain reaction（MSP）.The expression of 14-3-3σ protein was detected by Western blot.Viability of SGC-7901/CDPP cells treated with 5-Aza-CdR was observed by MTT assay.The cell cycle and apoptosis were analyzed by flow cytometry.Results Before 5-Aza-CdR treatment,14-3-3σ gene was hypermethylated and 14-3-3σ protein expression was not detected in SGC-7901/CDDP cells.Treated with 5,10 μmol·L-1 of 5-Aza-CdR for 72 h,hypermethylated 14-3-3σ gene was demethylated,so that 14-3-3σ protein was re-expressed and cisplatin resistance was reversed.Meanwhile,5-Aza-CdR could inhibit cell viability,increase the cell number in G0/G1 phase and induce the cell apoptosis in SGC-7901/CDDP cells in a dosedependent manner.Conclusion 5-Aza-CdR possesses inhibitory effects on the methylation of 14-3-3σ gene in SGC-7901/CDDP cells,and the resistance mechanisms of cisplatin are related with methylation of the 14-3-3σ gene.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2010年第7期918-921,共4页
Chinese Pharmacological Bulletin
基金
国家重点基础研究发展计划(973计划)前期研究专项(No2009CB526405)
