去甲基化制剂对HepG2细胞RUNX3基因表达及细胞生物学行为的影响

基础医学与临床 ›› 2007, Vol. 27 ›› Issue (3): 314-318.

去甲基化制剂对HepG2细胞RUNX3基因表达及细胞生物学行为的影响

张雪妍张煦

兰州大学基础医学院病理学教研室兰州大学基础医学院病理学教研室

收稿日期:2006-09-26 修回日期:2006-11-01 出版日期:2007-03-25 发布日期:2007-03-25
通讯作者: 张煦

Effect of demethylating agent on RUNX3 expression and the growth of human hepatocarcinoma cell line HepG2

Received:2006-09-26 Revised:2006-11-01 Online:2007-03-25 Published:2007-03-25

摘要/Abstract

摘要： 目的探讨人肝癌细胞系HepG2经5-氮杂-2'-脱氧胞苷（5-Aza-2'- deoxycytidine , 5-Aza-CdR）处理后诱导高甲基化失活的RUNX3基因重新表达的可能性及对细胞生长的影响，寻找抗癌治疗的新靶点。方法 RT-PCR检测抑癌基因RUNX3 mRNA的表达；MTT、集落形成实验观察细胞的生长活性；流式细胞术和透射电镜分析细胞周期及细胞凋亡的变化。结果肝癌细胞经不同浓度之5-Aza-CdR处理后，原无RUNX3 mRNA表达的细胞均检出基因重新表达，细胞生长速度出现不同程度减慢及细胞克隆形成率显著降低（P<0.01）。用药后肝癌细胞发生明显的S期阻滞，电镜显示肝癌细胞形态学改变。结论去甲基化制剂5-Aza-CdR能有效地激活肝癌细胞系HepG2因高甲基化所致RUNX3基因沉默的再转录，诱导该基因的表达，从而抑制肿瘤细胞生长。

Abstract: Objective This study was designed to investingate the effect of 5-Aza-2'-deoxycytidine (5-Aza-CdR)on cell growth and explore the possibility of re-expression of the hypermethylated and silenced RUNX3 gene in the hepatocarcinoma cell line HepG2. Methods The change in expression of the tumor suppressor gene RUNX3 mRNA in cultured HepG2 cells was observed by using RT-PCR before and after 5-Aza-CdR treatment. Activity of cell growth was observed by MTT assay and colony-forming test . The cell cycle was analyzed by flow cytometry . Apoptotic morphology was observed by transmitting electron microscopy. Results The gene was reactivated by two different doses of 5-Aza-CdR treatment in HepG2 cell not expressing RUNX3. The hepatocarcinoma cell line treated with 5-Aza-CdR displayed a slowed growth rate in contrast to the control group. The colony formation rate of HepG2 cell treated with 5-Aza-CdR decreased dramatically (P<0.01). The S phase was arrested and morphosis was changed as induced by 5-Aza-CdR. Conclusion 5-Aza-CdR may effectively cause demethylation and inhibit the growth of the hepatocarcinoma cell line HepG2 by reactivating the RUNX3 gene transcription silenced by aberrant hypermethylation.

张雪妍张煦. 去甲基化制剂对HepG2细胞RUNX3基因表达及细胞生物学行为的影响[J]. 基础医学与临床, 2007, 27(3): 314-318.

. Effect of demethylating agent on RUNX3 expression and the growth of human hepatocarcinoma cell line HepG2[J]. Basic & Clinical Medicine, 2007, 27(3): 314-318.