| Objective:To develop the rational DLL4cDNA vaccine to renal cell carcinoma and the mechanism of the vaccine on nude mice RCC models, we have thus developed the study to explore the possible immunization by DLL4-encoding plasmid DNA vaccine to evaluate the vaccine efficacy by some nude mice admitted to the RCC. We investigated the tumor local reactions which immunized by DLL4cDNA vaccine and blank group to provide some evidences for RCC immune therapy and clinic adjuvant therapy.Method:1.The development of DLL4-encoding plasmid:Using some specific primers to get the DLL4express box by PCR, we had a restrictive analysis, inserted pVAX1plasmids and did sequencing verification, purificated the transfected cells and had a Western bolt to carry out the DLL4cDNA clone.2. Mouse immunization:The pVAX1-DLL4plasmids were injected into the4-week male nude mice as an experimental group, comparatively, we have control group injected negative pVAX1plasmids.3. RCC node mice model and vaccine efficacy evaluation:RCC786-0cell line was injected into experimental group and none towards control group, we observed the circumstances of tumor growth in two group.Results:1. The DLL4cDNA expression box has a length of2058bp. Sequence verification and restricted enzyme cutting sites were proved after it was transfected into PVAX1plasmids.2. Tumor appeared in blank group3weeks later after786-0cell line had adopted. The average diameters were (1.26±0.91) cm. It proved that nude mice could be used as RCC animal models.3. Injection of PVAX1-DLL4plasmids could not stop tumor growth totally, but inhibited tumor growth since the tumor was not as big as that of control group. The average diameters of the tumor formed in mice injected PVAX1-DLL4plasmids were (1.07±0.70) cm and appeared red.Conclusion:1. PVAX1can be used as an efficient vector to deliver DLL4cDNA into mice to produce antibody to DLL4, which works as a vaccine to treat renal cell carcinoma.2. Nude mice can be used as a RCC animal model. |
PDF Full Text Request