Biological Character Of Melanoma B16 Cell Clones Mutagenized By Space Environment

Objective1. To explore the oncogenesis of the melanoma B16 cell clones mutagenized by space Environment,and find the B16 cell clones with remarkably mutated immunogenicity.2. To find the cell clones which were modified in cytoskeleton or surface structure.3. To determine the cell growth characteristics and observe the difference of biological character between the mutated cell clones and the control cell clone.4. To search the genes related to development and metastasis of melanoma.Methods1. B16 cells were carried by the Chinese 20^th recoverable satellite to the outer space, after 18 days' spaceflight and then monocloned. Four cell clones, which were randomly selected from the total 110 clones obtained, and the control clone were routinely cultured. The cultured cells were injected to 10 groups of C57BL/6J mice,8～11 mice in each group, five groups of mice received hypodermic injection and another five groups of mice received abdominal injection. The survival time were observed in abdominal injection groups. The mice in hypodermic injection groups were sacrificed after 14 days, the toumors, spleens and thymus were weighted,and the serum IL-2 concentration were determined. Moreover, the melanoma tumor tissues were examined histopathologically.2. The mutated cell clone and control B16 cells were detected with immuno-fluorescence cytochemistry and confocal lasers canning microscopy.3. The distribution of cell cycle of the mutated cell clones and the control cell clone were detected by FCM.4. The mutated cell clones and the control cell clone were imaged by AFM,and the difference of the image were observed between the images.5. The growth characteristics of the mutated cell clones and the control cell were detected by MTT assay.6. Gene chips were used to detect the gene expressions of the cell lines (8^#) which...