抗内甲基转移酶单克隆抗体的制备及临床应用研究

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O^6-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胞株，采用免疫组织化染色方法，检测60例脑瘤组织中MGMT表达水平，结果：得到了7株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60例脑瘤组织中有27例MGMT阴性，经亚硝脲药物治疗，其中24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，25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：建立了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MGTM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。

Preparation and clinical application of the monoclonal antibodies against the human O~6 -methylguanine-DNA methyltransferase

Objectives:To establish hybridomas secreting monoclonal antibodies(McAbs) against O 6 methylguanine DNA methyltransferase(MGMT) and to observe the relationship between MGMT expression and clinical responses to ACNU and BCNU in human brain tumors.Methods:The hybridomas were established by cell fusion.MGMT expression in 60 glioma specimens was detected by means of immunohistochemical assay.Results: Seven hybridomas secreting McAbs against MGMT were obtained.Thirty tumor specimens had no detectable or low level of MGMT expression(Mer -), while 30 specimens had high level of MGMT expression(Mer +). The Mer - patients showed more sensitive to ACNU and BCNU than the Mer + patients.Conclusions: The high specific hybridomas secreting monoclonal antibodies(McAbs) against MGMT were established.The preliminary study indicated that MGMT negative tumors were sensitive to ACNU and BCNU, whereas MGMT positive ones were more resistant to nitrosourea drugs.