Suppressing Proliferation And Inducing Apoptosis Effect Of Lentiviral Vectors Expressing Small Interfering RNA Targeting CD47Gene On Hep-2Cell

Objective To assess the effect of the lentiviral vectors expressing smallinterfering RNA (siRNA) for CD47gene knockdown in inhibiting Hep-2cellgrowth and apoptosis, to provide an effective target for gene therapy about laryngealcarcinoma.Methods After build lentiviral vector,a siRNA directed against CD47gene wastransfected into Hep-2cells; Cell morphological changes were observed withfluorescence microscope; The changes of CD47mRNA expression were detected bysemi-quantitive RT-PCR; The protein of CD47was evaluated by means of Westernblotting; The effect of suppressing proliferation of Hep-2cells was detected by MTTassay;phagocytosis test in vitro detectioned macrophages that the influence of Hep-2cells after CD47silence.Result After CD47-siRNA lentiviral plasmid transfected into laryngeal carcinomaHep-2cells, The morphology result showed the CD47siRNA to be able effectively tosuppress the Hep-2cell multiplication, the cell deformation was discovered, Thetumor cell volume was significantly smaller, the necrosis and apoptosis of tumor cellswere found under fluorescence microscope. The results of semi-quantitive RT-PCRand Western blotting: the expression of mRNA and protein of CD47decreasedsignificant in Hep-2cells (P<0.05), decreased the expression of CD47mRNA inHep-2cells by76%-82%, and decreased the expression of protein in Hep-2cells by77%; Resulting also increased cell apoptosis by MTT assay, when lentiviral vectorstransfectioned cells after48hours (P<0.01);The phagocytosis of macrophage toCD47-siRNAgene silencing of Hep2cells up.Conclusion The lentiviral vectors expressing CD47siRNA can significantly inhibitCD47gene expression in Hep-2cells and induce the cell apoptosis in vitro. Themacrophages improved the phagocytosis with CD47-siRNA gene silencing of Hep-2cells;The lentiviral vectors expressing CD47siRNA gene therapy might be a newstrategy in targeting molecular therapy of cancer of larynx, this data can also supportthe potential application of RNA interference to treat the laryngeal squamouscarcinoma.