摘要
目的:观察5-氮杂-2'-脱氧胞苷(5-aza-2'-deoxycitydine,5-Aza-CdR)对体外培养的胃癌SGC7901细胞和BGC823细胞增生、细胞周期和凋亡的影响及其对此两株细胞中Apaf-1基因的甲基化状态的影响.方法:不同浓度5-Aza-CdR处理体外培养的SGC7901细胞和BGC823细胞后,用MTT法检测处理24,48和72h的细胞增殖活性;PI染色和流式细胞仪检测药物处理后72h细胞周期分布和细胞凋亡率;MSP法检测用药前后细胞中Apaf-1基因的甲基化状态;RT-PCR法及Westernblot法检测用药前后细胞中Apaf-1的mRNA及蛋白表达的变化.结果:1×10-7,5×10-7,1×10-6和5×10-6mol/L5-Aza-CdR处理SGC7901和BGC823细胞24,48,72h后,细胞增生受到抑制,有时间和剂量的依赖性.流式细胞仪分析表明,各药物浓度处理72h后凋亡率增加明显:SGC7901细胞5×10-7,1×10-6和5×10-6mol/L组分别为2.53%±1.19%,5.93%±0.86%,10.14%±1.51%,与对照组(0.12%±0.03%)相比差异显著(P<0.05);BGC823细胞5×10-7,1×10-6和5×10-6mol/L组分别为1.57%±0.26%,4.64%±1.05%,8.21%±1.46%,与对照组(0.57%±0.03%)相比差异显著(P<0.05).在5-Aza-CdR处理前未检测到SGC7901细胞系及BGC823细胞系的Apaf-1基因的mRNA及蛋白表达,经过5-Aza-CdR处理后,Apaf-1基因在SGC7901细胞系及BGC823细胞系中甲基化状态得到了逆转,Apaf-1基因的mRNA及蛋白重新表达.结论:5-Aza-CdR对SGC7901细胞和BGC823细胞具有增生抑制作用;Apaf-1基因的表达情况与其甲基化状态的改变有关.
AIM： To observe the effects of 5-aza-2＇ -deoxycitydine （5-Aza-CdR） on the proliferation of human gastric cancer cell lines SGC7901 and BGC823 as well as the methylation and epression of Apaf-1 gene.
METHODS： Human gastric cancer cell lines SGC7901 and BGC823 were cultured in RPMI1640 and then treated with different concentrations of 5-Aza-CdR （1 ×10^-7, 5 × 10^-7, 1 × 10^-6 and 5 ×10^-6mol/L）. The proliferation of the cells was detected by MTT assay and flow cytometry （FCM）. The methylation of Apaf-1 gene in the two kinds of cell lines was detected by methylation-specific-polymerase chain reaction （MSP）, and the expression of Apaf-1 was detected by reverse transcription-polymerase chain reaction （RT-PCR） and Westen blot analysis.
RESULTS： 5-Aza-CdR displayed a growth inhibitory effect on SGC7901 and BGC823 cells in a dose-and time-dependent manner after exposure to 5-Aza-CdR at different concentrations （1 ×10^-7, 5 × 10^-7, 1 × 10^-6, 5× 10^-6 mol/L） for 24, 48 and 72 hours. FCM analysis showed that the apoptosis rates in SGC7901 cells （2.53% ± 1.19%, 5.93% ± 0.86%, 10.14%±1.51%） and BGC823 cells （1.57% ± 0.26%, 4.64%± 1.05%, 8.21% ± 1.46%） were increased significantly after exposure to 5-Aza-CdR （5 × 10^-7, 1×10^-6, 5× 10^-6mol/ L） for 72 hours as compared with 0.12% ± 0.03% and 0.57% ± 0.03% in the control cells （P 〈 0.05）. The methylation and loss of Apaf-1 mRNA and protein expression were detected in SGC7901 and BGC823 cells before 5-Aza-CdR treatment. However, the methylation was reversed and Apaf-1 was re-expressed after 5-Aza-CdR treatment. CONCLUSION： 5-Aza-CdR can inhibit the pro- liferation of SGC7901 and BGC823 cells through blocking cell cycles and inducing cell apoptosis, during which the reversion of Apaf-1 gene methylation plays an important role.
出处
《世界华人消化杂志》
CAS
北大核心
2007年第3期221-227,共7页
World Chinese Journal of Digestology
关键词
5-AZA-CDR
APAF-1基因
胃癌
甲基化
5-aza-2＇-deoxycytidine
Apaf-1 gene
Gastric cancer
Methylation
Cell apoptosis
Cell proliferation
