RNAi抑制食管癌细胞Eca-109中OGT的表达研究

RNAi inhibitting the expression of OGT in esophagel cancer cells Eca-109

目的:研究RNAi表达载体对食管癌细胞Eca-109中OGT基因表达的抑制作用。方法:构建针对OGT基因的干扰载体,用质粒转染食管癌细胞Eca-109,RT-PCR分析干扰载体对OGT基因的抑制情况,Western blot分析干扰后OGT蛋白的表达情况。结果:OGT基因在食管癌细胞Eca-109中高表达,三个干扰序列中瞬时转染干扰载体RNAi-OGTB-Eca109的干扰效率最高,可达80%。稳转干扰OGT蛋白水平抑制率65.21%,与对照组相比有统计学意义(P<0.05),O-GlcNAc水平下降(RL2反映)率37.5%,与对照组相比有统计学意义(P<0.05)。结论:用RNAi表达载体可以有效抑制食管癌细胞Eca-109中OGT基因的表达。

Objective：To investigate the inhibitory effects of RNAi expression vector on the expression of OGT in esophagel cancer cell Eca - 109. Methods：The expression of OGT in esophagel cancer cell Eca - 109 was examined with RT - PCR and Western blot. At the same time, the RNAi expression vectors aimed OCT were constructed and transfected into Eca - 109 cells with plasmids. The expression of OGT mRNA was detected with RT - PCR and the expression of OGT protein was detected with Western blot. Results：There was a high expression of OGT in Eca - 109 ceils. The expression of OGT gene was inhibited about 80% by RNAi - OGTB in transient transfection Eca - 109 ceils. The expression of OGT protein was inhibited about 50.6% in stable transfection Eca - 109 cells ,and there was a significant difference compared with control group （P 〈 0.05 ）. The level of O- GIcNAc decreased 37.5% in the interfered group compared with control group and there was significant difference （ P 〈 0.05 ）. Conclusion： The RNAi expression vector can effectively inhibit the expression of OGT gene in esophagel cancer cell Eca- 109 cells.