关键词: 短发夹状RNA, 核表达载体, 化生长因子-β1

Abstract:

Objective To construct the plasmid containing short hairpin RNA（shRNA） of TGF-β1 expression vector． Methods Short chain oligonucleotide was designed according to the TGF-β1 mRNA sequence provided by Genebank, then DNA segment was gained through annealing after chemosynthesis, and then was cloned to pWH1 vector. The recombinant TGF-β1 shRNA expression vector was evaluated by using enzyme cutting. At last, the constructed TGF-β1 expression vector was transfected into salivary gland mucoepidermoid carcinoma（Ms） cells by Lipofectomine TM 2000, and its effect on TGF-β1 expression was observed by RT-PCR and immunohistochemistry. Results Successful construction was identified by enzyme cutting and the constructed plasmid was called pWH1-TGF-β1. The shRNA and it inhibited the TGF-β1 mRNA and protein expression effectively. Conclusion The constructed TGF-β1 shRNA expression vector can block the TGF-β1 expression in salivary gland mucoepidermoid carcinoma cells.

Key words: shorthairpinRNA, ukaryoticexpressionvector, ransforminggrowthfactor-β1