RNAi抑制NF-κB基因增强肺癌细胞对TRAIL敏感性的研究

Study of NF-κ B gene repressed by RNAi on enhancing the sensitivity of lung cancer cells to TRAIL

目的:探讨RNAi靶向沉默NF-κB基因对TRAIL诱导肺癌细胞凋亡的影响。方法:使用NF-κBp65siRNA转染肺癌A549细胞48h,实验分为空白对照、脂质体对照以及干扰实验3组。采用RT-PCR法测定肺癌A549细胞内NF-κBp65mRNA的表达,MTT法检测siRNA转染前后TRAIL对A549细胞生长抑制作用的变化。结果:与空白对照和脂质体对照组相比,siRNA组具有明显抑制肺癌A549细胞NF-κBp65mRNA表达的作用(P<0.01)。MTT实验表明,转染siRNA后TRAIL对A549细胞的生长抑制作用明显增强,但在同一浓度,转染NF-κBp65siRNA的细胞与未转染组相比,细胞增殖活性明显下降(P<0.05)。结论:应用RNAi技术可有效干扰NF-κBp65的表达,抑制肺癌A549细胞的增殖,并可增加肺癌细胞对TRAIL的敏感性。

Objective： To investigate the effect of NF-kB siRNA-mediated gene silencing on the sensitivity of lung carci- noma cells to TRAIL. Methods： NF-kB p65 siRNA was used to transfect lung carcinoma A549 ceils for 48 h. The experiment included 3 groups： The untreated cell group （blank control group）, the liposome-treated cell group （lipo- fectin group） and the siRNA transfection group （the interference experimental group）. The expression of NF-gBP65 gene was detected by RT-PCR method, and the inhibition of A549 cell growth by TRAIL before and after the trans- fection of siRNA were assayed by MTT method. Results： Compared with the blank control group and the lipofectin group, siRNA transfection significanly inhibited the expression of NF-kB P65mRNA in lung carcinoma cells A549 （P 〈0. 01）. MTT experiment indicated that the inhibition of TRALT to A549 was rernarkbly enhenced after siRNA trasfection, but in the same concentration, compared the tranfection ceils of NF-~B p65 siRNA with those in the un- transfection group, the activity of the cell proliferation obviously decreased （P〈0.05）. Conclusions ： The application of RNAi techniqe can effectively inhibit the expression of NF-kB P65 and the proliferation of the lung carcinoma cell A549. What＇s more, it may greatly increase the sensitivity of cancer ceils to TRAIL.