摘要
目的:研究siRNA对生长抑素(SOM)基因表达的抑制作用．方法:根据生长抑素基因序列设计和合成 RNA干扰的靶序列,应用RiboMAXT7体外转录试剂盒合成siRNA并转染胃癌细胞系 SGC-7901,经RT-PCR,免疫组化法检测生长抑素在mRNA和蛋白质水平的抑制效应．结果:成功合成了siRNA(21 bp)．胃癌细胞系 SGC-7901中SOM基因的表达呈强阳性,分布在胞质中,siRNA作用48 h后SOM基因的表达明显抑制,与空转染和空白对照组相比有显著性差异(49．71％±0．056％vs 10．49％±0．021％, 0％,P<0．01)．结论:siRNA可特异性抑制生长抑素基因的表达．
AIM： To research inhibitory effects of small interfering RNA （siRNA） on the expression of somatostatin gene.
METHODS： According to the gene sequence of somatostatin in GenBank, we designed the siRNA-targeted templates and synthesized siRNA using T7 RiboMAX Express RNAi System in vitro. The obtained siRNA was then transfected into gastric cancer cell line SGC-7901. Reverse transcription polymerase chain reaction （RTPCR） and immunohistochemical techniques were used to detect the expression of somatostatin at both mRNA and protein levels.
RESULTS： The target siRNA with a length of 21 bp was successfully synthesized. Before transfection, somatostatin was strongly and positively expressed in gastric cancer cell line SGC-7901, locating at the cytoplasm. Forty-eight hours after transfection, somatostatin expression was markedly inhibited and the inhibitory rate in siRNAtransfected cells was significantly higher than that in the cells transfected with empty vector and non-transfected cells （49.71% ± 0.056% vs 10.49% ± 0.021%, 0%, both P 〈 0.01）.
CONCLUSION： siRNA can inhibit the expression of somatostatin specifically.
出处
《世界华人消化杂志》
CAS
北大核心
2006年第8期784-788,共5页
World Chinese Journal of Digestology
