TP53基因改变影响MDM2抑制剂对神经胶质瘤的效应及机制分析-药物分析学专业论文.docxVIP

ABSTRACT Background and objective Glioma is one of the most common malignant tμMors in central nervous system. Malignant gliomas are characterized by its invasiveness and dissemination, resulting in frequent tμMor recurrence after surgical resection and/or conventional chemotherapy and radiation therapy. Current combinations of surgical therapy, radiotherapy and chemotherapy regimens do not significantly improve long-term survival of the patients with malignant glioma. The abnormal behaviors demonstrated by cancer cells are the result of a series of mutations in key regulatory genes. The cells become progressively more abnormal as more genes become damaged. The transition from a normal, healthy cell to a cancer cell is step-wise progression that requires genetic changes in several different oncogenes and tμMor suppressors. The growing awareness of differences in inter-individual response to radiochemical exposure calls for a new approach in the therapy and follow-up of cancer patients. P53 acts as a tμMor suppressor in many tμMor types, induces growth arrest or apoptosis depending on the physiological circμMstances and cell type. TP53 is frequently mutated or inactivated in about 60% of cancers. P53 is ubiquitinated by MDM2, which leads to proteasomal degradation. The goal is to improve the therapy results of Nutlin-3 by evaluating inter-individual differences in TP53 status. Methods Glioma cell lines U87, A172 and U251 were used to evaluate the effects of MDM2 inhibitor Nutlin-3. First, the whole coding sequence of the p53 gene was cloned from cDNA by RT-PCR and sequenced. Second, the MTT assay was used to measure the activity of cell proliferation. Third, the effects of combination of Nutlin-3 and PI3k inhibitor, LY294002, were detected. Fourth, after the drug treatment, the mRNA levels of p53 and the target genes of p53, MDM2, P21 and BAX were compared with the control by semi-quantitative RT-PCR. Fifth, Western-blots were used to detect the changes of p53, PARP