Pre-clinical Study on Hepatocellular Carcinoma Therapy by Targeted Gene Drug (ASOR-PLL-hIL12)
|School||Central South University|
|Keywords||Targeted therapy vectors ASOR-PLL Human interleukin- 12 HepG2 cells month Conformational Expression|
Objective: To effectively carry out liver cancer gene therapy , the use of targeted therapy vectors asialo mucin - poly- L- lysine (ASOR-PLL) exogenous gene of human interleukin 12 (hIL12) into HepG2 cells in order to understand hIL12 expression and influencing factors , initially explore solutions targeting gene expression inefficient way drugs . Method : first construct pcDNA3.1 () / P40, pcDNA3.1 (-) / P35 and two subunits of expression pcDNA3.1 () / IL12 three kinds of plasmids , gene transfection angle determined from the ASOR-PLL target vector isotropic , the three kinds of plasmids bind composed two kinds of targeted genetic medicine [ASOR-PLL-P () / IL12 and ASOR-PLL-P () / P 40 , ASOR-PLL- TEM gene targeting drugs at different concentrations of the molecules excipients conformation . Targeted gene in different configurations drugs transfected HepG2. According to different expression by semi- quantitative RT-PCR, ELISA, Western blot method screened conducive to the expression of the best molecular conformation . Results: 1 . Therapy vectors ASOR-PLL cells on ASGr targeting ; 2 . pcDNA3.1 () / P40, pcDNA3.1 (-) / P35 and two subunits of expression pcDNA3.1 () / IL12 three kinds of plasmids can be transferred and expressed in HepG2 cells hIL12 protein molecules, but liposomal dyeing and ASOR-PLL transfection results to differ ; 3 . Concentrations of two different materials targeting gene drug molecule conformation different drug molecules of different configurations after transfection, the expression level varies 25 ~ 150nm in diameter annular granular or highest expression . Conclusions : 1. Therapy vectors ASOR-PLL and dual subunit coexpression pcDNA3.1 () / IL12 plasmid prepared in this study is a treatment for liver cancer targeting gene drugs ; 2 . Gene targeting drug molecule conformation exogenous genes have important implications ; 3 . Exogenous gene fragment size and connectivity also have some impact on its expression .