Construction of IGF-II RNA Interference Lentiviral Vector and Establishment of Its Stably Transfected Cell Line

Objective To construct the IGF-Ⅱ RNA interference lentiviral vector and establish the stably transfected human adrenocortical tumor cell line SW-13.Methods According to the IGF-Ⅱ gene sequence,three shRNA targeting IGF-Ⅱ were designed,and the corresponding pLLU2G-shIGF recombinant plasmids were constructed.After the most effective shRNA was achieved,the shuttle plasmid pLLU2G-shIGF with lentiviral packaging plasmid was mixed,and 293FT cells were transfected.Virus solution was collected to infect SW-13 cells,and the stably transfected cell line was established.Results The third shRNA sequence targeting IGF-Ⅱ showed the best gene silencing effect as 80%,and the stable transfected cell line was constructed successfully based on this sequence,and the inhibition ratio was 95% for IGF-Ⅱ mRNA.Conclusion It is successful to obtain IGF-Ⅱ-shRNA lentiviral vector that can effectively down-regulate the expression of IGF-Ⅱ in human adrenocortical tumor cell line SW-13 and establish the stably transfected cell line.