Construction,expression and examinations of recombiant plasmid with pmRNA IRES-hKDR / 中国基层医药

Objective To construct recobinant pmRNA IRES-hKDR and lay a primary foundation for further study of gene therapy for tumors.Methods pmRNA IRES-hKDR was obtained from pDC520 hkDR and pmRNA IRES-Luc which was reserved by our laboratory through molecular biology technology and was transfected into Hepall-6 via lipsome after identification of PCR,restriction enzyme digesting and DNA ELISA and Western blot.Results The recombiant plasmid had pmRNA IRES-hKD gene which was expressed in Hepall-6//G418 in vitro and the expressed protein was secreted out of the cells could response with specific hKDR antibody,which was identified by Western blot.Conclusion An eukaryotic expression recombiant plasmid pmRNA IRES-hKDR can be constructed successfully which provides the possibility of further researches on its anti-tumor effect.

Immunity protection of kinase domain-containing receptor fusion gene vaccine to melanoma models in murine / 中国医师杂志

Objective To investigate the immune protection of dendritic cells(DCs) harboring kinase domain-containing receptor(KDR) fusion gene on mice carrying B16 melanoma.Methods The bone marrow precursor-derived dendritic cells(BMDCs) were induced from bone marrow progenitors of mice by GM-CSF.The KDR fusion gene mRNA was transfected into DCs in vitro.Mice were immunized with the same amount of DCs at 7-day interval and then each mouse was injected with 5?10~5 B16 cells.The mice without tumors were injected with B16 cells again 20 days later.Mice were randomly divided into 4 groups: group A(n=7),group B(n=8),group C(n=8) and group D(n=7).The mice were immunized with DCs one time in group A,2 times in group B,3 times in group C and as controls in group D.Results After one week,all mice in group D had tumors with average survival 15 days.All mice in group A,B and C had no tumors after 20 days later.After the second injection of B16 cells,2 mice in group A and 2 mice in group B had tumors.The mice in group C had no tumor.The average survival periods were calculated from first injection of B16 cells to the study end.The average survival period of group A was 50 days and that of group B was 72 days.Conclusions The dendritic cells vaccine harboring KDR fusion gene has immune protection against melanoma in mice.