脱氧核酶对人食管癌细胞增殖的影响

目的 :探讨针对增殖细胞核抗原 (PCNA)基因起始密码AUG的脱氧核酶 (DNAzyme)对人食管癌细胞(EC970 6 )增殖的影响。方法 :设计合成DNAzyme ,应用脂质体转染法将其转入体外培养的hEC。采用流式细胞仪测细胞周期 ;四甲基偶氮唑蓝 (MTT)比色法分析hEC增殖活性。采用免疫组化 (SABC法 )检测PCNA蛋白表达。结果 :2 .0 μmol/LDNAzyme干预EC970 6 3d后 ,MTT比色吸光度值低于对照组 (P <0 .0 1)和反义寡聚核苷酸 (ASODN)组 (P<0 .0 5 )。DNAzyme和ASODN对吸光度值的抑制呈剂量依赖性。细胞干预 2d后 ,DNAzyme和ASODN均减少细胞核中棕黄色颗粒的表达 ;DNAzyme、ASODN和对照组的G0 /G1期细胞的比率分别为 72 .5 %、6 6 .8%和 5 6 .7%。结论 :针对PCNA的DNAzyme能调控细胞周期 ,有效抑制EC970 6的体外增殖。

The Effect of DNAzyme on the Proliferation of Human Esophageal Carcinoma Cells

Objective To investigate the inhibi tr oy effects of DNAzyme to human proliferating cell nuclear antigen gene on the p roliferation of human esophageal carcinoma cells(EC9706) in vitro.M ethods The DNAzyme specific to PCNA gene was designed and synthesize d .Using lipofectamine mediated method,the DNAzyme was infected on EC9706.The e ffects on cell proliferation were examined by MTT assay and cell cycle was detec ted by flow cytometry. The level of PCNA protein was detected by immunohistochem ical technique.Results The A value by MTT colorimetric an alysis of 2.0 μmol/L DNAzyme group was lower than that of control group(P<0.01)and antisense oligodeoxynucleotide (ASODN) group( P<0.05)at 3 days after transfection, respectively. The decrease of A va lue of DNAzyme group and ASODN group showed a dose-dependence within the experim ental range at 3 days. After 2 days, the percentage of quiescent cell (G 0/ G 1) were 72.5%、66.8% and 56.7%, respectively in DNAzyme, ASODN and contro l groups, DNAzyme and ASODN could downregulate the expression level of PCNA prot ein .Conclusion These data implicate that DNAzyme specific to PCNA could regulate cell cycle and could be a useful method to suppress the proliferation of EC9706 effectively.