| Glioma is among the deadliest of tumors: Most of the 20,000 people diagnosed each year in the United States with this form of brain cancer die within 2 years. The tumors, which derive from support cells of the brain known as glia, are eerily invasive: Glioma cells spread freely into healthy surrounding brain tissue, making it impossible to remove all of the tumor surgically and virtually assuring that the cancer will soon return elsewhere in the brain. Neurosurgeons have been searching new drugs and approaches to fight against it for many years. Now, chemotherapy is still an important way to treat the disease. But the drugs that are used have too many side effects. It is urgent to find other drugs, which are sensitive to the glioma cells to deal with those problems.It was reported that Antigliomatin has the inhibitory effect on glioma cell. However, there is no reportion about the effect of Antigliomatin on cell morphogeny. After uranyl acetate staining, the change of ultrastructure of C6 cell before and after Antigliomatin treatment was observed under transmission electron microscopy. This study aimed to: 1.observe the inhibitory effect of Antigliomatin on C6 glioma cells. 2.describe the change of morphology after the treatment of Antigliomatin. 3.test the expression of p16 protein by immunohistochemistry. 4.discuss the able mechanisms of the inhibition on the glioma cell.First, we identified the C6 glioma cell by detecting the expression of Glial fibrillary acidic protein(GFAP). Our result showed that there were many brown granules in the cytoplasm and nucleus of tumor cells. It was proved that the cells were C6 glioma cell. So we could do the next experiments with our cells.The inhibitory effect of Antigliomatin on the glioma cellAfter the subculture of C6 glioma cells, we divided the cells into experiment group and contrast group. Treating the experiment group cells with Antigliomatin of different concentrations or Cisplatin for 72 hours, then we observed its cytotoxicity under phase contrast microscope, hematoxylin and eosin staining and transmission-scanning electron microscope; At the same tine, we also tested their killing effect by Mononuclear cell direct cytotoxicity assay (MTT method). Our results showed that in the groups treated with Antigliomatin or Cisplatin, there were reduced number of cells sticking to the walls of the culture flask and more round cells under light microscope; irregular, pycnosic and reduced nuclei and swollen mitochondria under electron microscope. MTT results showed that Antigliomatin had apparent cytotoxic effects on C6 glioma cells. Furthermore, with the concentration of Antigliomatin increasing, the cytotoxic effects became more apparent. When the concentration of Antigliomatin was 64ng/L, its inhibitory rate was 59.68%. So we concluded that Antigliomatin had apparent cytotoxic effects on C6 glioma cell and induced apoptosis to C6 cells. The influence of Antigliomatin on cyclin protein p16Gene p16 also called multiple suppressor gene (MST), localizes in 9p21, and it is an important gene associated with cell cycle. The protein p16 encoded by gene p16 has relationship with the control of cell cycle. As an inhibitive factor of cyclin dependent kinase-4, protein p16 is able to inhibit cell division. And in many glioma cells, researchers have found the defection of the expression of protein p16, which indicates that protein p16 has close relationship with brain glioma. In this paper, we tested the expression of protein p16 before and after the treatment of C6 glioma cells with different concentration of Antigliomatin. The result showed that after the treatment, the expression of protein p16 rose obviously. In the experiment group, we could see brown granules in cytoplasm. In contrast with control group, the control group cells had statistics difference.It is concluded, therefore, that Antigliomatin is capable of inducing apoptosis of C6 glioma cell and inhibit the tumor growth, which can establish important theoretical basis...
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