摘要
为观察RNA干扰对过表达Hlrg蛋白的Hep G2细胞的影响,构建了封闭Hlrg基因表达的shRNA表达载体pAVU6+27-Hlrg,将pAVU6+27-Hlrg稳定转染到过表达Hlrg蛋白的Hep G2细胞中;分别用相差倒置显微镜、FCM、SABC-FITC、RT-PCR、Western blot等观察细胞改变。测序结果表明pAVU6+27-Hlrg构建成功;针对Hlrg的RNA干扰已完成;相差倒置显微镜发现细胞突起增加;FCM显示干扰组G1期明显缩短。针对Hlrg的RNA干扰能促进过表达Hlrg蛋白的Hep G2细胞的生长。
The influence of RNA interference targeting on the over-expression of human lipopolysaccharide response gene （Hlrg） on Hep G2 cells was investigated, in which the expression vector pAVU6（ ＋ ）27 Hlrg that could block the expression of the Hlrg gene was constructed, andsubcloned into shRNA vector pAVU6（＋）27. Then the pAVU6（＋）27-Hlrg plasmid was trasfected to Hep G2-pcDNA3. 1 （＋）-Hlrg cell line by means of trasfection reagent. The changes in the trasfected cells were examined by contrast inverl microscopy, FCM, SABC-FITC, RT-PCR and Western blotting analysis. It was demonstrated that the plasmid pAVU6（ ＋ ）27-Hlrg was successfully constructed and the RNA interference targeting againt H lrg had been completed. As demonstraled by invert microscopy, the cellular processes increased and the FCM analysis revealed the remarkable shortening of the Gl phase in the RNA interference group. It is concluded the RNA interference targeting can improve the growth and proliferation of Hep G2 cells in which the human lipopolysaccharide response protein is over expressed.
出处
《现代免疫学》
CAS
CSCD
北大核心
2006年第4期274-277,共4页
Current Immunology
基金
国家自然科学基金资助项目(30170361
30400413
30471675)
第四军医大学博士后基金资助项目(00001149)
