| MAN2C1 is a human alpha-mannosidase encoded by a cDNA cloned in our lab, which we originally called 6A8 alpha-mannosidase. This enzyme has been internationally named mannosidase, alpha, class 2C, member 1[Homo sapiens], abbreviated as MAN2C1. The relevant gene locates at chromosome 15q11-13. Committee of Human Gene Nomenclature(HUGO) has named the gene MAN2C1. MAN2C 1 has a molecular weight of～116 KDa in deglycosylation state.In order to study of the biological significance of MAN2C1, Man2c1 transgeneic ICR mice were developed in this laboratory by microinjection of linear pIRES2-EGFP-MAN2C1. Integration of Man2c1 gene into the genomic DNA was examined by genomic PCR in this thesis. Eight mice were found to be genomic PCR positive in 113 original mice.～1/2 F1,～3/4 F2 and～80-90 F3 mice were PCR positive. In 7 lines of the mice examined, 4 were RT-PCR positive for Man2c1 mRNA, indicating that Man2c1 transgeneic ICR mice had been developed. The hair becomes Phenotypic change was observed on the transgeneic mice number 35., whose hair have become sparse in comparing to that on the wild type ICR mice. Effect of the transgene on the growth of transplanted tumor and on huumoral immune response was studied in this thesis. In the experiment of transplanted tumor, growth of tumor was found faster in the transgeneic mice than in the wide type mice. Furthermore, tumor in the transgeneic mice invaded to the surrounding tissues and metastasized to draining lymph nodes. In contrast, there was complete capsure on the tumor in the wild type mice. The data suggest a promotive effect of MAN2C1 on tumor growth. In study of the mechanism related, number of NK cell in the spleen of the transgenic mice was found to be higher than that of the wild type mice. However, cytotoxic effect of the splenocytes on YAC-1 cells was not changed. In the experiment of humnoral immune response, mice were immunized with BSA. The spleens in the transgenic mice were enlarged much more than those in the wild type mice. However, level of anti-BSA antibody in the serum of the transgenic mice was lower than that of the wild type mice.In order to further study the biological significanse of MAN2C1, expression of Man2c1 in mouse fibroblast cell NIH-3T3 was inhibited by means of RNAi technique. Effect of inhibition of Man2c1 gene expression on growth of the cell in vitro was examined. However, no positive effect was observed. |
