| Objective: Glioblastoma is a kind of highly malignant intracranial tumor.At present,the effect of conventional surgery and radiotherapy and chemotherapy is not satisfactory.Methods: 1.Establishment of animal model: The method of establishing in situ glioma model is that the C6 glioma cells were implanted into the right caudate nucleus of SD rats with stereotaxis(Midline right 3mm,coronal suture forward 1mm,deep 5mm).2.Vaccine preparation: The anti-glioma vaccine is a mixture of allogeneic 9L glioma cells,allogeneic 9L glioma cell lysates,and syngeneic C6 glioma cell lysates.3.Experimental grouping:100,000U-dose treatment group(n = 23),200,000U-dose treatment group(n = 23),control group(n = 23),sham group(n = 23).4.Vaccine therapy: The vaccine treatment time was eighth days after modeling.The 100,000U-dose treatment group injected 100,000 allogeneic 9L glioma cells,100,000 allogeneic 9L glioma cell lysate,100,000 syngeneic C6 glioma cell lysate;The 200,000U-dose treatment group injected 200,000 allogeneic 9L glioma cells,200,000 allogeneic 9L glioma cell lysate,200,000 syngeneic C6 glioma cell lysate;control group and sham group were injected with 3ml PBS.5.Survival time record: Observation of the starting point for the end of modeling surgery,the end point was seventieth days after modeling.The rats were injected with pentobarbital overdose in dying condition(to euthanasia),survival time was recorded after euthanasia.6.Tumor volume analysis: All rats were examined by MRI after modeling 7th,14 th,21th,28 th,35th and 70 th day.The tumor volume was calculated by ImageJ software and Cavalieri ’s formula.7.Pathological examination: The brain tissue of rats(dying under the condition)were removed for HE staining.8.Glioma invasion factor detection: CD147,MMP-2,Fascin,P75,and Podoplanin were selected as invasive factor related antibodies.On the eighteenth day after modeling,3 rats was randomly selected from each group and the brain tissues were stained with immunohistochemistry.9.Statistical analysis: All the data analyzed with SPSS software(17.0.0).The results of tumor volume were expressed as mean ± SD(standard deviation)and tested with independent sample T-Test.The results of positive cell rate were expressed as mean ± SD and tested with analysis of variance(ANOVA).The results of survival time were analyzed using Kaplan-Meier estimator analysis and compared using the log-rank test.The above tests were performed to check the difference between different groups and p<0.05 was set as level of significant difference.Results: 1.Survival time: The survival time of 100,000U-dose treatment group was longer than that of 200,000U-dose treatment group(P < 0.05)and control group(P < 0.05).The survival time of 200,000U-dose treatment group was longer than that of control group(P < 0.05).The survival time of rats in sham group was more than 70 days.2.Tumor volume: On the 14 th day and the 21 th day after the operation,the tumor volume of 200,000U-dose treatment group was less than that of 100,000U-dose treatment group(P < 0.05)and control group(P < 0.05),and the tumor volume of 100,000U-dose treatment group was less than that of control group(P < 0.05).There was no tumor growth in sham group.3.Pathological examination: the tumor of 100,000U-dose treatment group had subsided and no residual tumor cells were found.The tumor cell density of 200,000U-dose treatment group was lower than that of the control group.The tumor cells were not observed in sham group.4.Glioma invasion factor detection: in the comparison of positive cells of CD147,MMP-2,Fascin,p75,Podoplanin,200,000U-dose treatment group was lower than 100,000U-dose treatment group(P < 0.05)and control group(P < 0.05),100,000U-dose treatment group was lower than control group(P < 0.05),the sham group almost no positive staining.Conclusion: Glioma vaccine based on molecular mimicry could be used to treat intracranial glioma.The therapeutic effect was enhanced with the increase of dose,however,excessive doses may lead to side effects such as brain edema or nerve damage. |
