论文部分内容阅读
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。
Objective: To investigate the effect of DNAzyme targeting AUG of PCNA gene on the proliferation of human esophageal cancer cells (EC970 6). Methods: DNAzyme was designed and synthesized and transfected into hEC cultured in vitro by lipofection method. The cell cycle was measured by flow cytometry. The proliferative activity of hEC was analyzed by MTT assay. The expression of PCNA protein was detected by immunohistochemistry (SABC method). RESULTS: The MTT colorimetric absorbance of 2.0 μmol / L LDNAzyme was lower than that of control (P <0.01) and antisense oligonucleotide (ASODN) groups (P <0.05) ). The inhibition of absorbance by DNAzyme and ASODN was dose-dependent. DNAzymes and ASODN decreased the expression of brown granules in the nucleus after 2 days of intervention. The ratios of DNAzyme, ASODN and G0 / G1 phase cells in the control group were 72.5%, 66.8% and 56.7% . Conclusion: DNAzyme targeting PCNA can regulate the cell cycle and effectively inhibit the proliferation of EC9706 in vitro.