摘要
目的研究5-氮杂-2′-脱氧胞苷(5-Aza-CdR)对OS-RC-2肾癌细胞株增殖、凋亡的影响。方法用不同浓度10-7、10-6、10-5、10-4mol/L的特异性DNA甲基转移酶抑制剂5-Aza-CdR处理OS-RC-2肾癌细胞株,未经药物处理细胞作对照(对照组)。透射电镜观察5-Aza-CdR处理OS-RC-2肾癌细胞株前后细胞形态学变化。MTT检测OS-RC-2肾癌细胞株抑制率。流式细胞仪检测OS-RC-2肾癌细胞株凋亡。以甲基化特异性PCR(MSP)检测细胞处理前后γ-catenin基因的甲基化状态。结果用药后细胞器肿胀、线粒体空化、溶酶体增多,核质不均匀。5-Aza-CdR明显抑制OS-RC-2肾癌细胞的增殖,而且与药物浓度及作用时间在一定范围内成正相关(P<0.05)。10-7,10-6,10-5mol/L5-Aza-CdR处理细胞后,细胞凋亡率增高[(3.74±0.34)%,(7.85±0.59)%,(12.93±1.32)%vs.对照组(0.86±0.08)%],成剂量依赖性(P<0.01);作用3d后,在10-6mol/L时细胞周期中处于G0/G1期的显著增多(P<0.05),细胞周期阻滞于G0/G1期。未经5-Aza-CdR处理的OS-RC-2细胞中γ-catenin基因启动子区域高甲基化,经5-Aza-CdR10-5mol处理72h后,γ-catenin基因启动子区域高甲基化得到逆转。结论5-Aza-CdR能消除γ-catenin基因启动子甲基化状态,使其重新表达而抑制OS-RC-2肾癌细胞株的生长,使细胞周期阻滞于G0/G1期,并促进其凋亡。
Objective To study the influence of 5-Aza-2＇-deoxycytidine （5-Aza-CdR）on the proliferation and apoptosis of human renal carcinoma OS-RC-2 cell line. Methods Human renal carcinoma OS-RC-2 cells were treated with a cocentration of 10^-7 , 10^-6 , 10^-5 or 10^-4 mol/L 5-Aza- CdR,respectively. The growth rate of the ceils was detected by MTT assay and ultrastructural changes were observed under transmissional electron microscope. The apoptosis was analyzed by flow cytometry. Results 5-Aza-CdR inhibited the proliferation of OS-RC-2 cells in a time-and concentration-dependent manner （P〈 0. 05 ）. There were significant increases in apoptotic rates of OS-RC-2 cells [（3. 74±0. 34）%,（7.85±0. 59） %, （12.93±1.32）% vs. （0. 86±0. 08）%]（P〈0. 01） after 5-Aza-CdR treatment. The cells number of G0/G1 phase increased at a concentration of 10^-6 mol/L（P〈0. 05）,and the cell cycle was stopped at G0/G1 phase after treated for 3 days. The methylation status of γ-catenin gene promoter region was reversed with 5-Aza-CdR 10^-5 mol/L treating for 72 h. Conclusion 5-Aza-CdR can inhibit the growth of OS-RC-2 cells and promote their apoptosis by elimilating demethylation of γ-catenin gene CpG-rich promoter regions.
出处
《江苏医药》
CAS
CSCD
北大核心
2009年第6期708-710,I0002,共4页
Jiangsu Medical Journal
基金
江苏大学临床医学科技发展基金(JLY20050015)
