反义甲基转移酶基因片段增强肿瘤坏死因子相关凋亡诱导对肝癌细胞的作用

Impacts of the transfection of antisense DNA methyltransferase Ⅰ gene fragment into hepatocellular cell on its sensitivity to tumor necrosis factor related apoptosis inducing ligand

目的 观察转入反义甲基转移酶 (DNMT) 1基因片段后的肝癌SMMC 772 1细胞对肿瘤坏死因子相关凋亡诱导配体 (TRAIL)的敏感性改变。方法 使用脂质体法将真核表达载体 pCl neo转入肝癌SMMC 772 1细胞内 ,PCR法鉴定载体的转入 ,锥虫蓝排斥试验检测活细胞率 ,末端脱氧核苷酰转移酶介导的dUTP缺口末端标记法 (TUNEL法 )检测细胞凋亡率。结果 PCR法证明载体成功转入肝癌SMMC 772 1细胞内 ,转入反义DNMT1基因片段的肝癌SMMC 772 1细胞与转入正义DNMT1基因片段和空载体的细胞相比 ,活细胞率显著降低 (P <0 .0 5 ) ,凋亡率显著增高 (P <0 .0 5 )。结论 转入反义DNMT1基因片段的肝癌SMMC 772 1细胞对TRAIL的敏感性明显增强。

Objective To observe the impacts of the transfection of antisense DNA methyltransferase Ⅰ(DNMT1) gene fragment into SMMC 7721 cell on the changes of cell sensitivities to tumor necrosis factor related apoptosis inducing ligand(TRAIL). Methods The eukaryon expression vector pCl neo was transfected into SMMC 7721 cells by liposomes and the transfection was identified by PCR. Survival cell rate was measured by trypan blue exclusion, apoptotic rate was determined by in situ TdT dUTP terminal nick end labeling(TUNEL) method. Results PCR detection indicated that the eukaryon expression vector pCl neo was successfully transfected into SMMC 7721 cells. Survival cell rate of SMMC 7721 cells transfected with antisense DNMT1 gene fragment was remarkably lower than that of transfected with sense DNMT1 gene fragment and empty vector ( P <0.05 or 0.01 ), but the apoptotic rate was significantly higher ( P <0.05 or 0.01).Conclusion The sensitivity of SMMC 7721 cells to tumor necrosis factor related apoptosis induced ligand can be enhanced by the transfection of antisense DNMT1 gene fragment.