| Background and ObjectiveGlioblastoma is a common central nervous system tumors derived from epithelial tissue, are malignant, with invasion of surrounding tissue, invasive growth, no capsule, ill-defined, easy to relapse after abscission. Hitherto, glioblastoma is still no way to cure thoroughly. The surgery were used primarily, with radiotherapy, chemotherapy, biological and immunity auxiliary treatment on clinical. Currently, realgar in the application of anti-tumor limited to the treatment of blood system carcinoma, curative effect and achieved a higher response rate. But the reporte related research has not been found about the effect of realgar on the central nervous system carcinoma.The cell culture in the life sciences has become an important technology. Under the invivo studies with cell ultrastructure means will be studied in depth the proliferation of cell, the molecular mechanism of apoptosis provides new clues and tools., It was found that the incidence of many diseases are associated with disorders related to apoptosis with in-depth research on apoptosis. Apoptosis has become the one of the most striking areas in the life sciences at now. Bcl-2 is the most in-depth study and extensive family of apoptosis genes. Bcl-2 inhibits apoptosis and Bax that it is another member of the apoptosis family was promoting apoptosis.This topic take came from the human glioblastoma tissue first to isolated from tumor tissue. Then through examines Bcl-2 and the Bax two gene around realgar intervention change preliminary discussion realgars in the anti-tumor aspect molecular mechanism.Materials and methodsThe specimens of nine glioblastoma are all obtained by surgical abscission from April to December 2010 in Neurosurgery Department of the Second Affiliated Hospital of Zhengzhou University (All specimens were authenticated on postoperative pathologic test). All the tissue were all primary cases without preoperative chemotherapy and radiation therapy. After tumor resection surgery, the appearance of a good selection of tumor tissue stored in the pre-made in the culture fluid kept at 0℃-4℃. Immediately, tumor tissue were separated and cultured at post-operation. In accordance with the 2007 WHO Classification Norm of Central Nervous System Tumors, nine samples are all glioblastoma. First of all, the tumor cells were isolated from tumor tissue by cold trypsin. Then, all specimens use paste the method which the wall raises, with the cell in the logarithm vegetal period an its transfer of generation, intervenes its growth by the realgar solution. The realgar solution concentration divides into the low concentration(40mg/L) and the high potency(70mg/L) two groups. The RT-PCR was used separately before and after the intervention to examinate the Bcl-2 gene and Bax gene mRNA in the glioblastoma cell's expression quantity and carries on the contrast around the realgar intervention. SAS9.1 statistical software was used, and significant level is:α=0.05.Results1. Giemsa staining results:Through the microscope,the phenomenon can be seen that the obvious cell nucleus assumes the violet, the nucleolus blue black, the cytoplasm shines diaphanous, and the obvious cell and the cell nucleus shape, the size vary, and presents the binuclear and the polynuclear phenomenon, in the morphology tallies with the tumour cell.2. RT-PCR Results:The volume of Bcl-2 mRNA before and after intervention of realgar were 0.89±0.04,0.46±0.05(40mg/L),0.48±0.03(70mg/L); The volume of Bax mRNA before and after intervention of realgar were 0.35±0.11,0.85±0.07(40mg/L),0.88±0.11(70mg/L). The mRNA expression of Bcl-2 and Bax were statistically significant differences before and after intervention with realgar. But the differences between different concentrations were not significant.ConclusionRealgar induced apoptosis of human glioma cells in the mechanism of apoptosis-related gene may increase anti-apoptotic gene Bax and decreased Bcl-2 expression. |
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