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目的 :建立抗人甲基转移酶单克隆抗体细胞株并建立相应的蛋白测定方法。观察脑瘤组织中O6 甲基鸟嘌呤 DNA甲基转移酶 (MGMT)表达水平与亚硝脲药物疗效的关系。方法 :采用细胞融合技术建立抗人甲基转移酶单克隆抗体细胞株 ,采用免疫组织化学染色方法 ,检测 6 0例脑瘤组织中MGMT表达水平。结果 :得到 7株稳定分泌抗人甲基转移酶单克隆抗体的杂交瘤细胞株 ,并建立了测定MGMT免疫组化方法。观察的 6 0例脑瘤组织中有 2 7例MGMT阴性 ,经亚硝脲药物治疗 ,其中 2 4例好转 ,3例复发 ;另 3例MGMT可疑阳性的患者 ,经亚硝脲药物治疗 ,其中 2例好转 ,1例复发 ;2 5例MGMT阳性患者 ,经亚硝脲药物治疗 ,5例好转 ,12例复发 ,8例死亡 ;5例MGMT为强阳性患者 ,使用亚硝脲药物治疗无效 ,全部死亡。结论 :建立的 7株稳定分泌抗人甲基转移酶单克隆抗体的杂交瘤细胞株及MGMT蛋白测定方法 ,为临床开展亚硝脲预见性化疗提供了必要手段。初步结果显示 ,脑瘤组织中MGMT表达水平与亚硝脲药物疗效及预后有关 ,是肿瘤细胞对亚硝脲药物产生耐药性的基础
Objective: To establish a monoclonal antibody against human methyltransferase cell lines and to establish the corresponding protein assay. To observe the relationship between the expression level of O6-methylguanine DNA methyltransferase (MGMT) and the efficacy of nitrosourea in brain tumor. Methods: Monoclonal antibody against human methyltransferase was established by cell fusion technique. The expression of MGMT in 60 cases of brain tumor was detected by immunohistochemical staining. Results: Seven hybridoma cell lines stably secreting monoclonal antibodies against human methyltransferase were obtained and the immunohistochemical method for the determination of MGMT was established. Twenty-seven cases of MGMT were negative for MGMT, of which 24 cases were improved and 3 cases were relapsed after treatment with nitrosourea, while the other 3 cases were suspected of positive MGMT treated with nitrosourea, 2 cases improved, 1 case relapsed; 5 cases of MGMT positive patients treated with nitrosourea drugs, 5 cases improved, 12 cases of recurrence, 8 patients died; 5 cases of MGMT strong positive patients, the use of nitrosourea drugs ineffective, All died. Conclusion: The established seven hybridoma cell lines stably secreting anti-human methyltransferase monoclonal antibody and MGMT protein assay provide the necessary means for clinical prospective chemotherapy of nitrosourea. The preliminary results show that the expression level of MGMT in brain tumor is related to the curative effect and prognosis of nitrosourea drugs and is the basis of drug resistance of tumor cells to nitrosourea drugs