Construction and efficacy identification of the lentiviral vector harboring RNAi based on the hyperplasia suppressor gene (HSG).

e22177 Background: Our previous study suggested that hyperplasia suppressor gene (HSG) induced apoptosis of mitochondrial pathway via MAPK signal transduction pathway. In order to study on the apoptosis mechanisms via mitochondrial pathway induced by the HSG gene on human lung adenocarcinoma cells, we constructed and identified the efficacy of a lentiviral vector harboring RNA Interference sequence targeting the HSG gene. Methods: Three siRNA sequences targeting the HSG mRNA were designed. The levtivirus vectors of GCSIL-GFP-HSG were constructed and confirmed by DNA sequencing. The 293T cells were co-transfected with pGCSIL-GFP-HSG, pHelperl.0 and pHelper2.0 in order to produce the virus stocks. The titer of the virus was also tested. The lentivirus was transfected to human A549 lung adenocarcinoma cells. The expression of the HSGgene was analyzed by Real-time PCR. Results: DNA sequencing suggested that the DNA sequences were consisted with it designed, which meant that the RNAi sequence targeting the hum...