Suppression of the expression of hPOT1 gene in human gastric cancer cell line SGC-7901 with vector-based RNA interference technique / 解放军医学杂志

ABSTRACT

Objective To construct the expression vector of siRNA targeting the human protection of telomeres (hPOT1) gene for the observation of the inhibitory effect of the vector on hPOT1 gene expression in gastric cancer cell line SGC-7901, and to offer a ground work for further study on the roles of hPOT1 in growth and proliferation of gastric cancer cells. Methods 64 base-pair oligonucleotide for small interfering RNA expression targeting hPOT1 gene was designed and chemically synthesized. After annealing, double oligonucleotides were inserted into the downstream of H1 RNA promoter of pSUPER to recombine psiRNA-hPOT1 plasmids, and transiently transfected into gastric cancer cell line SGC-7901 with liposome-mediated transfection after restriction enzyme and sequencing analysis. Semi-quantitative RT-PCR was used to detect the expression levels of hPOT1 gene. Result Recombinant psiRNA-hPOT1 vector was confirmed by sequencing analysis. The results demonstrated that 64nt had been inserted into the expected site. Furthermore, the inserted sequence was exactly correct. The recombinant psiRNA-hPOT1 vector could significantly suppress the hPOT1 expression in gastric cancer cell line SGC-7901. Conclusion The constructed siRNA expression vector of hPOT1 gene psiRNA-hPOT1 recombinant plasmid can block the expression of hPOT1 gene in gastric cancer cell line SGC-7901.