| Objective: We proposed to develop a mouse model of metastasis of murine colon carcinoma cells and develop a murine colon carcinoma cellline of metastasis that were tranduced with IL-21 gene, further examine wheather the survival of murine colon carcinoma cell that have highly metastasis ability can be inhibited by the celline that can express IL-21 and have highly metastasis ability. Methods: 1 Establishment of mouse model of metastasis of murine colon carcinoma cell. Murine colonrectal carcinoma Colon26,a undifferentiated adenocarcinoma, was transplanted into subcutaneous tissue of homogenous Balb/c murine inner pod. After forming a tumor masses in pod, popliteal fossa lymphonodus was obtained in asepsis condition, then granulated it into granule through a net which have 400 holes, grounded tissue was developed in RMPI1640 medium supplemented with 10% fetal calf serum. therefore, tumor cellline that have metastasized into lymphonodus could be obtained in the medium. Then monocelled clone of the metastatic tumor cell was obtained,further it's tumor speciality was demonstrated by immunohistochemistry of CEA and was named Colon26M. Rate of lymphonodus metastasis of the Colon26M celline was evaluated and compared with the rate of lymphonodus metastasis of the Colon26 celline. 2 Establishment of murine colon carcinoma Colon26 celline that can express IL-21 gene. Colon26M cellline that can express IL-21 gene, named Colon26M/IL-21, was obtained by the retrovirus vector LXSN which have carried IL-21 DNA through ecotropic ψ2 cell and amphotropic PA317 cell transfected conlon26 cell. The RNA of colon26M/IL-21 cellines was isolated by the direction of TRIzol Reagent.IL-21DNA was amplificated according to the isolated RNA by RT-PCR methods, and DNA band analysis of the amplificated products was travaled by gel scanner. The RNA and DNA of the Colon26 cellline was obtained by the same way and as a control. 3 Animal experiments Colon26M/IL-21 cellline was transplanted into murine single inner pod of each mouse as a experiment group, and Colon26M celline as a control, tumor growth of each transplanted murine pod was observed,and the tumor volume was evaluated every other 48 hour.On the 25th day after Colon26M/IL-21 and Colon26 cellline were transplanted into each pod ,rate of lymphonodus metastasis were evaluated and compare between experiment group and control group.The spleen cell of each mouse transplanted with Conlon26M/IL-21 was cultured with Colon26M/IL-21,the production of IFN-γwas examined by ELISA method, spleen cell of each mouse transplanted withConlon26M was cultured with Colon26M as a control. Result: 1 Establishment of mouse model of metastasis of murine colon carcinoma cell. Colon26 celline that have metastasized into lymphonodus has been isolated from only one murine popliteal fossa lymphonodus successfully, tumor special CEA of the celline was demonstrated by immunohistochemical method,and the celline was named Colon26M. 2 Establishment of murine colon carcinoma Colon26M celline that can express IL-21 gene:Colon26 celline that can express IL-21 gene was established by retrovirus vector LXSN which carried IL-21 cDNA,and it was named Colon26M/IL-21.Both bright 28sRNA bands of Colon26M/IL-21 and Colon26M were observed in gel scan showed that both general RNA had been isolated. It was demonstrated that the Colon26M/IL-21 celline can express IL-21mRNA by special DNA band on 434bp mark had seen in DNA products gel scan which had obtained from amplication by RT-PCR method. 3 Animal experiment: There was tumor growth in each murine pod that had transplanted with Colon26M/IL-21 cell,then the tumor regressed spontaneously, but there was developing tumor growth in each murine pod that had transplanted with Colon26M cell. Lymphnodus metastasis rate of Colon26M/IL-21 cell was 0%, Lymphonodus metastasis rate of Colon26M was 50%,statistic demonstrated there had been significative difference between the experiment and control group by exactprobabilities in 2×2 table(α=0.05, P=0.0866) . Spleen cell in experiment group secr...
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