The Treatment Of Murine Pancreatic Carcinoma By β-elemene Combined With Immune Response Induced By Dendritic Cells Modified With Interleukin 23

Objective To study the pathogenesis of pancreatic carcinoma and investigate the p53 genes expression in carcinogenesis and development of pancreatic carcinoma induced by DMBA. Methods High dose (10mg/100gweight) of dimethylbenzanthracene (DMBA) was put into the parenchyma of mouse pancreas to develop a model of pancreatic carcinoma in mice. Immunohistochemical SP method and was used to study the expression of P53 in pancreatic carcinoma induced by DMBA. Results The prevalence of pancreatic carcinoma among DMBA-treated mice within 3-5 months was 30% ( 12/40 ) . 3 mice developed pancreatic ductal adenocarcinomas with glandular duct-like distribution of cancer cells, various sizes of glandular cavities, obvious nucleiheterotpic and typical cocus of cancer cells under the microscope. There was no expression of P53 in normal pancreatic tissues. The overexpression of P53 was detected in pancreatic carcinoma (10/12). Conclusion High dose DMBA was put into the parenchyma of rat pancreas could obtained a pancreatic cancer model with high incidence in a short time. There are P53 protein overexpressions in the pancreatic carcinoma induced by DMBA in mice, and P53 protein overexpression may relate to the degree of malignancy of the tumors , the result obtained in this study was quiet similar with the human pancreatic cancer gene mutation types. Objective To study the method of obtaining a large number of dendritic cells. To study the specific CTLs effect against tumor cells initiated by dendritic cells pulsed with peptide of cancer cell . To offer the basic theory for the treatment of pancreatic cancer with DC vaccine combined with β-elemene. Methods Development of cells with cytologic features of dendritic cell in bone marrow cultures supplemented with GM-CSF and IL-4. determining the DC phenotype and the specific structure of DC by electronic microscopy. To observe the CTLs effect against pancreatic carcinoma leading by the Dendritic cell pulsed with tumor cell lysate in vitro. Results A large number of typical dendritic cells proliferated by supplementing with GM-CSF and IL-4 cytokines. Dendritic cell has specific cell appearance and structure , and highly express various cell surface molecules. rmTNF-αhas the ability of stimulating DC mature, the mature DC has the enhancing abilities of antigen presenting and IL-12 self-secreting ,as well as, express higher levels of CD54,MHC-II 及CD86 molecules than control group, P<0.05. T lymphoid cell stimulated by dendritic cells without tumor antigen could not recognize and kill the target cells , only if dendritic cells pulsed with peptide of cancer cell can lead a strong immune response to special tumor cells. The inhibiting ratio of CTLs is significant higher than that in other groups. Conclusion Bone marrow DC has strong ability of inducing special CTLs against determined cancer cells after they were pulsed with tumor cell lysate. DC vaccine is probably a new immunotherapeutic method against tumor in the near future.Objective To clone two subunits P19 and P40 of the Interleukin 23 gene, to construct the dual-expression vector pcDNA3-IL-23 in order to study the anti-tumor therapy inducing by the dendritic cells genetically transfect by Interleukin 23. Methods total RNA of P19 and P40 were extracted from thymus and spleen of mice respectively by one step way, and cDNA was amplified by RT-PCR, the PCR product was linked with pMD18-T vector and E.Coli JM109 was transformed by pMD18-P19 and pMD18-P40, positive plasmid was selected by PCR method. P19 and P40 gene were digested from pMD18-P19 and pMD18-P40, the product was inserted into pcDNA3.0 expression vector, selected positive plasmid was tested by XhoI/KpnI restrict enzyme. Result Total RNA of P19 and P40 from thymus and spleen respectively was complete, The sequence of cloned gene is consistent with the reported sequence of the Genebank. The constructed pcDNA3-IL-23 eukaryotic dual-expression vector was proved to have P19 and P40 subunits of 1.62kbp by restriction endonuclease analysis. Conclusion We successful...