5-氮杂-2'-脱氧胞苷对HepG2中抑癌基因MIA2表达的影响  

Effect of 5-Aza-2'-deoxycytidine on the expression of tumor suppressor gene MIA2 in HepG2

作　　者：杨少奇[1] 姚文暾[1] 杨花[1] 黄睿[1] 胡建国[1] 何芳[1] 

机构地区：[1]宁夏医科大学总医院消化内科,宁夏消化疾病研究所,宁夏银川750004

出　　处：《西安交通大学学报（医学版）》2013年第4期479-482,共4页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.81060194)~~

摘　　要：目的应用甲基化酶抑制剂5-氮杂-2'-脱氧胞苷(5-Aza-2'-deoxycytidine,5-Aza-CdR)作用肝癌细胞株HepG2,观察其对细胞增殖和凋亡的影响,并观察用药后肝癌细胞中黑色素瘤抑制蛋白2(melanoma inhibitory activity 2,MIA2)表达的变化情况。方法分别以浓度0、1、2、4、8、10、20、40μmol/L的5-Aza-CdR作用HepG2细胞,MTT法检测细胞增殖,流式细胞术检测细胞凋亡。再以0μmol/L和10μmol/L的5-Aza-CdR作用HepG2细胞,细胞免疫组化和Western blot检测细胞中MIA2蛋白表达。结果 5-Aza-CdR作用HepG2后,MTT结果显示细胞增殖受到抑制,在一定药物浓度范围内,细胞增殖抑制率和药物浓度呈正相关;流式细胞术结果显示细胞早期凋亡增加,在一定药物浓度范围内,早期凋亡率和药物浓度呈正相关;细胞免疫组化和Western blot结果显示与对照组(0μmol/L)比较,10μmol/L组中MIA2蛋白表达增强。结论 5-Aza-CdR作用HepG2细胞后,MIA2蛋白表达明显增强,细胞增殖受到抑制,早期凋亡增加。Objective To observe the effect of methylase inhibitor 5-Aza-2＇-deoxycytidine（5-Aza-CdR） on the proliferation and apoptosis of HCC cell line HepG2 and detect the expression of MIA2 protein in HepG2 after 5-Aza-CdR treatment.Methods The proliferation and apoptosis of HepG2 treated with 5-Aza-CdR of 0,1,2,4,8,10,20 and 40 μmol/L were detected by MTT and flow cytometry assay.Then after HepG2 was treated with 0 μmol/L and 10 μmol/L of 5-Aza-CdR,the MIA2 protein expression was detected by immunohistochemistry and Western blot.Results MTT showed that after treatment with a certain concentration range of 5-Aza-CdR,the proliferation of HepG2 was inhibited and there was a positive correlation between cell proliferative inhibition rate and drug concentration.Flow cytometry showed that the apoptosis of HepG2 was increased and a positive correlation existed between early apoptosis rate and drug concentration.Immunohistochemistry and Western blot revealed that the level of MIA2 protein was elevated in 10 μmol/L group compared with that in control group（0 μmol/L）.Conclusion After HepG2 is treated with 5-Aza-CdR,MIA2 protein expression is enhanced,cell proliferation is inhibited and early apoptosis is promoted.

关 键 词：5-氮杂-2'-脱氧胞苷 黑色素瘤抑制蛋白2(MIA2) 肝癌细胞 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]