5-Aza-CdR对肺癌SPC-A1细胞生长及p16基因异常甲基化的影响  

Effects of 5-Aza-CdR on proliferation of SPC-A1 lung cancer cell line and abnormal methylation of p16 gene

作　　者：马红兵[1] 白明华[1] 王宝峰[1] 任宏涛[1] 

机构地区：[1]西安交通大学医学院第二附属医院肿瘤科,西安710004

出　　处：《陕西医学杂志》2010年第10期1275-1278,共4页Shaanxi Medical Journal

基　　金：陕西省科技攻关基金资助(2004K13-G20)

摘　　要：目的:观察5-氮杂脱氧胞苷(5-Aza-2'-deoxycytidine,5-Aza-CdR)对体外培养的肺癌SPC-A1细胞增生、细胞周期及其对此细胞株中p16基因的甲基化状态的影响。方法:不同浓度5-Aza-CdR处理体外培养肺癌SPC-A1细胞后,用MTT法检测药物处理24、48、72h后的细胞增殖活性;PI染色和流式细胞仪检测药物处理72h后的细胞周期分布;MSP法检测用药前后细胞中p16基因的甲基化状态;RT-PCR法检测用药前后细胞中p16的mRNA表达变化。结果:2×10-6、5×10-6、1×10-5mol/L 5-Aza-CdR处理肺癌SPC-A1细胞24、48、72h后,细胞增生受到抑制,有时间和剂量依赖性。流式细胞仪分析表明,不同药物浓度处理肺癌SPC-A1细胞72h后细胞增殖指数降低明显:5×10-6、1×10-5mol/L组分别为(34.65±0.52)、(28.29±0.79),与对照组(43.85±1.17)相比,差异有统计学意义(P<0.05)。在5-Aza-CdR处理前未检测到肺癌SPC-A1细胞系的p16基因的mRNA表达,经过5-Aza-CdR处理后,p16基因在肺癌SPC-A1细胞系中甲基化状态得到了逆转,p16基因的mRNA重新表达。结论:5-Aza-CdR对肺癌SPC-A1细胞具有增生抑制作用;p16基因的表达情况与其甲基化状态的改变有关。Objective：To observe the effects of 5-Aza-2＇-deoxycytidine,（5-Aza-CdR） on the p roliferation of SPC-A1 lung cancer cell line as well as on the methylation and expression of p16 gene.Methods：Human lung cancer cell line were cultured in RPM I1640 and then treated with different concentrations of 5-Aza-CdR.The proliferation of the cells were detected by MTT assay and flow cytometry（FCM） after being cultured for 24,48 and 72h.The methylation of p16 gene in the cell line was detected by methylation specific polymerase chain reaction（MSP）,and the expression of p16 was detected by RT-PCR.Results：5-Aza-CdR displayed a growth inhibitory effect on SPC-A1 cell in a dose and time dependent manner after exposure to 5-Aza-CdR at different concentrations（2×10-6,5×10-6,and 1×10-5mol/L） for 24,48 and 72 hours.FCM analysis showed that the proliferation index（PI） in SPC-A1 cells（34.65±0.52,28.29±0.79） were decreased significantly after cell＇s exposure to 5-Aza-CdR（5×10-6,1×10-5mol/L） for 72 hours as compared with that（43.85±1.17） in control cells（P0.05）.The methylation and loss of p16 mRNA expression were detected in SPC-A1 cells before 5-Aza-CdR treatment.However,the methylation was reversed and p16 was re-expressed after 5-Aza-CdR treatment.Conclusion：5-Aza-CdR can inhibit the proliferation of SPC-A1 cells through blocking cell cycles and inducing cell apoptosis,during which the reversion of p16 gene methylation plays an important role.

关 键 词：肺肿瘤 基因 p16 DNA甲基化 细胞增殖 @5-Aza-CdR 

分 类 号：R375.2[医药卫生—病原生物学]