Antitumor Effects Of HN Gene Of Newcastle Disease Virus

In the previous part of this research, five nucleic acid expressing plasmids, pIRHNcc, pIRHNsp, pIRFcc, pIRFsp, pIRHNFcc were constructed by inserting different genes into pIRESlneo plasmid. Another nucleic acid expressing plasmid was constructed by inserting HN gene into pVAXl. The expressions of HN gene in Hela and OS732 cells were determined by hemagglutination tests, Western blot and indirect immunoflurescence assay. The results suggested that the hemagglutination titer of pVHN was highest,followed by pIRHNcc and plRHNsp,and the following was pIRHNFcc. Specific bright yellow-green fluorescence was observed on the membrane and across the plasma of the transfected cells that contained HN gene expressing plasmids when tested with anti-chicken IgY FITC conjugate.The assay of growth inhibition of cell, acrodine orange staining, cytoskeleton staining and genome DNA electrophoresis assay were applied to campare the induced changes of tumor cells transfected by recombined plasmids with that treated by NDV.The tumor models, C57BL/6 mice bearing B16 melanoma was successfully established in the left leg of mice by subdermal injection of tumor cells.Cs7BL/6 mice were implanted with 2x105 melanoma cells (B16),and then NDVcc and recombined plasmids were injected into tocaJ tumor at 2, 10 days after tumor growth to about 5mm in diameter. Minor clinical signs and lower metastatic carcinoma of the liver, kidney and spleen, the more apoptotic and necrosis cells, the more endoplasmic reticulum and secondary lysosome of tumor cells were observed in NDVcc,pVHNcc and pVHN+NDVcc group.These three groups had the higher CTL and NK. activity of tumor bearing mice,either.