The Anticancer Research Of Antisense Oligonucleotides Targeting Insulin-like Growth Factor 1 Receptor

Insulin-like growth factor 1 receptor(IGFlR) is over-expressed in many tumor cells. It is involved in cell proliferation and apoptosis and stimulates cell motility. Hence, targeting IGFIR provides a new way for cancer gene therapy. Here we mainly evaluated the antitumor effects of antisense oligonucleotide against IGFIR (IGF1R-AS) in vitro and in vivo, and the combined effects with chemotherapeutic drugs and irridiation. All these researches will promote the novel drug to clinical trail.Firstly, the inhibitory effect of IGF1R-AS on proliferation of twelve human tumor cells was estimated by MTS assay in vitro . In a 96 culture plate, 3,000⁴,000 cells were seeded in 100μl of the appropriate growth medium. When cells had grown to 50%-60% confluence, Lipofectin Reagent was used to transfect IGF1R-AS with different concentrations. 72 hours later, the MTS was added to cells. After 1¹.5 hours at 37℃ in a humidified, 5% CO₂ atmosphere, the absorbance at 490nm was recorded using an ELISA plate reader and the inhibitory rate was gained. The concentration-effect profile could be plotted and the IC₅₀ value could be determined using Logit analysis. The IC₅₀ of IGF1R-AS on twelve human cancer cells proliferation ranged between 0.122μmol/L⁰.477μmol/L. These findings strongly suggested that IGF1R-AS had wide antitumor activities in vitro. To investigate the specific activity, IGF1R-AS was transfected into HepG2 cells at three doses of 0.2,0.4,0.8μmol/L .Compared with sense and mismatched oligonucleotides, IGF1R-AS significantly inhibited HepG2 proliferation and IGF-1R expression at both mRNA and Protein levels, with respective inhibitory rate of 85.0%, 74.5% and 87.0% at the dose of 0.8μmol/L.Secondly, in vivo experiment, we used an orthotopic human hepatocellular carcinoma(HCC) model. 25 mg/kg/day, 50 mg/kg/day, 75 mg/kg/day of ASODNs, 50...