| Glioma is the most common tumor of brain, it has a higher prevalence accounting for35.26%-60.96%of intracranial tumors. Glioma has a wide range of invasive and progress from low level to high-level features and characteristics of the high rate of recurrence, high morbidity, high mortality, caused great harm to human health. Currently, the primary means of treatment of glioma is surgical resection with radiotherapy and chemotherapy, however, median survival in malignant glioma patients is less than two years. Taxol as an alkaloid class of drugs in the treatment of tumors of the clinical chemistry, has been widely used in breast cancer, ovarian cancer, brain tumors and tumor chemotherapy. Its unique anti-cancer mechanisms has attracted the attention of many experts and scholars, and may be used as expected glioma chemotherapy drugs or radiation sensitizing agent.Objective:This study aimed to explore the mechanism of PTX inhibition of glioma cell value-added and its application in malignant tumors of the nervous system to provide a theoretical basis.Methods:1. Screening cells:an initial period of the glioma cell line U373with, prepare the reagents and materials, read the relevant experimental literature and learn to be familiar with the method and the principle of which need a variety of styles.2. Cell culture:culture the cells grew well and be familiar with the growth cycle of U373cells, cell shape and characteristics.3. Detected by MTT cell viability determination of the value-added inhibition rate of the cells, and experimental grouping, then go to the following experiment.4. The cell cycle is Measured by flow cytometry distribution of the different experimental groups.5. Its morphological changes in different experimental groups were observed under fluorescence microscope by Application of an immunofluorescence technique.6. Extraction of RNA the in each group of cells, reverse transcription cDNA and amplification of the cell cycle gene DNA, and then apply the gel electrophoresis method for detection of changes in RNA levels of different experimental groups.7. Extract the group of cellular proteins, and detect cell cycle related protein expression level changes by Western Blotting.8. The statistical analysis software is application to give a statistically significant correlation processing.Results:1. PTX could inhibit cell proliferation on U373cells, and the inhibitory effects were in time and dose dependent manners.2. Cell cycle analysis showed that the G2/M phase of U373cells treated with PTX increased significantly compared with the control group.3. Immunofluorescence showed that the cell of experimental group were blocked in mitosis, increasing the stability of microtubules, the microtubule depolymerization obstacles.4. RT-PCR of results showed that PTX can increase the expression of the related RNA levels of the CDK1and CyclinB1, reduce the expression of the related RNA levels of CyclinD1, but has little effect on CDK2-related RNA.5. Western Blotting of results showed that PTX can increase the expression of the CDK1’s and CyclinBl’s protein levels, reduce the expression of the CyclinDl’s related protein levels,but has little impact on CDK2-associated protein.Conclusion:PTX can inhibit glioma proliferation of U373cells and cause cell cycle of mitotic arrest, Lead to apoptosis,the mechanism is closely related to its cell cycle related protein expression level. |
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