转染NF-κB抑制物基因对食管癌细胞系生物活性的影响

The Studies on Inhibitory Action of Transfection of NF-κB Suppressor Gene for Bioactivity of Esophageal Carcinoma Cell Line

目的:将NF-κB抑制物基因pcDNA3/mIκBαS32A/S36A转染食管癌细胞株EC1,观察对NF-κB因子活性及对细胞生长的影响。方法:采用脂质体转染法将pcDNA3/mIκBαS32A/S36A质粒转染入EC1细胞进行瞬时表达,检测其对EC1细胞的影响,用Westernblot和RT-PCR方法检测转染前后NF-κBp65、NF-κBp50蛋白和mRNA表达水平的变化。结果:与转染pcDNA3.0质粒和未转染质粒组比较,转染pcDNA3.0/mIκBαS32A/S36A质粒EC1细胞的生长速度受到明显抑制,P<0.05。转染pcDNA3/mIκBαS32A/S36A质粒的EC1细胞,在0、24、48、96小时未见NF-κBp65、NF-κBp50蛋白和mRNA表达,而转染pcDNA3.0质粒EC1细胞可见核内NF-κBp65和NF-κBp50蛋白表达。结论:转染NF-κB抑制物基因IκBα后可抑制EC1细胞的生长,并抑制细胞NF-κBp65、NF-κBp50基因的表达,该结果提示调节NF-κB的表达可能是食管癌基因治疗的靶点。

Objective： To transfect the EC1, the esophageal cell line, with NF-kB suppressor gene pcDNA3/mIlBαS32A/S36A and to observe the activity of NF-kB and its effect of cell growth. Methods： The lipidosome infection protocol was used to transfect the pcDNA3/mIkBαS32A/S36A plasmid into the EC1 cells for transient expression and detect the effect of the method on EC1 cells. The change in the expression level of NF-kB p65 and NF-kB p50 protein and mRNA before and after transfection was detected using the methods of Western blot and RT-PCR. Results： Compared to the transfected and the non-transfected pcDNA3.0 plasmid group, the growth velocity of the EC1 cells for transfection of the pcDNA3.0/mIkBαS32A/S36Aplasmid was apparently inhibited, P〈0.05. There was no expression of the NF-kB p65,NF-kB p50 protein and mRNA in the EC1 cells for transfection of the pcDNA3.0/mIkBαS32A/S36A plasmid within 0, 24, 48 and 96 hours but there was expression of the intranuclear NF-kB p65 and NF-kB p50 protein and mRNA in the EC1 cells for transfection of the pcDNA3.0/mIkBαS32A/S36A plasmid. Conclusion： After transfection of NF-kB suppressor gene IkBα, the growth of EC1 cells and expression of NF-kB p65 and NF-kB p50 cytogene were inhibited. It suggests that regulation of NF-kB expression may be the target of gene therapy for esophageal carcinoma.