IL12修饰的小鼠乳腺癌细胞EMT6IL12抗肿瘤用研究.docx

Interleu Interleu kin．1 2 modified mammary sarcoma murine cell line reveals anti-tumor effect ABSTRACT OBJECTIVE：To obtain the IL一12 modified mammary sarcoma murine cell line EMT6／IL-·1 2 which could highly express hlL·-1 2 and investigate the anti-·tumor effect of EMT6／IL—l 2 in mice model． METHODS：Recombinant eukaryotic plasmid encoding IL一1 2 gene(pcDNA6一p70)WaS transfected into EMT6．The positive clones were selected and identified．30 mice were randomly divided into 3 groups：EMT6／IL-1 2，pcDNA6一p70 and EMT6．Mice in EMT6／IL一1 2 group were inoculated S．C in the right prothorax with EMT6／IL—I 2；then．on day 4，7，1 O，l 4，1 7 after the inoculation，saline were injected on the primary injected site． Mice in the pcDNA6．p70 and EMT6 groups were inoculated with EMT6 cells and then pcDNA6一p70，saline were injected respectively in the same way as that of EMT6／IL一1 2 group．All mice were sacrificed on day 2 1．Subcutaneous tumors，spleens，thymus glands were weighted and the capacities of anti—tumor immunity including cytotoxicity, proliferation of spleen lymphocytes，the serial IFN—Y level and the frequencies of CD4+／IFN-丫and CD8+／IFN—Y double positive cells in spleen lymphocytes which stimulated by ConA were also investigated． RESULTS：EMT6／IL一12 could express hlL-12 in a high level．The inhibitory rate oftumor in EMT6／IL-1 2(97．08％)is higher than that of pcDNA6-p70(76．25％)and EMT6 (萨0．O 1)．Moreover,the capacities of anti—tumor immunity such as cytotoxity and proliferation of spleen lymphocytes，level of serial IFN-丫were also enhanced in EMT6／IL-1 2 group pO．o i)．Frequencies of CD4+／IFN吖，CD8+／IFN一丫double positive cells in spleen lymphocytes were increased significantly in EMT6／IL一1 2 group (CD4／IFN-Y：7．49士O．05％；CD8／IFN-Y：6．3 5+0．06％)compared to that of groups pcDNA6-p70(CD4／IFN-Y：4．3士0．03％；CD8／IFN—Y：3．7+0．05％) and EMT6 (CD4／JFN—y：2．05+0．04％；CD8／IFN-Y：1．26-士0．03％)∞O．01)． CoNCLUSIoN：EMT6／IL一12 cell line which highly express hIL一12 were acquired and the recombinant IL一1 2 modified mammar