人多发性骨髓瘤裸鼠皮下移植瘤模型的建立

Establishment of Human Multiple Myeloma Transplantation Tumor Nude Mouse Model by Injecting Subcutaneously RPMI8226

目的：探索人多发性骨髓瘤裸鼠皮下移植瘤模型建立的方法。方法：Balb／c裸鼠进行3Gyx线照射预处理后24h，于其皮下接种2×10^7人多发性骨髓瘤细胞RPMI8226，每2—3d观察裸鼠体重的变化、肿瘤生长的情况、裸鼠的生存时间等，待裸鼠濒死或死亡，或者超过观察时间仍未死亡时，处死裸鼠，取其皮下结节及器官进行病理检测，同时摘眼球取血行血清免疫固定电泳检测。结果：荷瘤裸鼠出现皮下结节的高峰为荷瘤后第2～3周，结节体积基本达到高峰为荷瘤后第5～6周；裸鼠死亡高峰出现在荷瘤后第7周，荷瘤小鼠中位生存时间为荷瘤后52d；裸鼠处死后血清中未检测到人源单克隆免疫球蛋白；皮下结节HE染色证实瘤组织为浆细胞来源，器官HE染色显示肝脏部分炎性损伤。结论：采用RPMI8226人骨髓瘤细胞于Balb／c裸鼠皮下接种（接种细胞数为2×10^7个细胞）造模的方法，具有操作过程较简单、技术要求低，观察肿瘤生长较为直观等优点；同时采用X线照射预处理的方法可降低裸鼠免疫力，明显提高模型的成瘤率；人多发性骨髓瘤细胞RPMI8226皮下接种于裸鼠，肿瘤生长过程中很难有人源单克隆免疫球蛋白分泌入血；荷瘤裸鼠出现肝脏的损伤，也许可作为今后抗肿瘤药物疗效研究的指标之一。

Objective To establish the nude mouse model of human multiple myeloma （MM） transplantation tumor by injecting subcutaneously RPMI8226, a human MM cell line. Methods 2 ×10^7 RPMI8226 cells were subcutaneously injected into the nude mice which had been exposed to the radiation of 3 Gy X-ray for 24 hours before the performance. The body weight, survival time of mouse and the situation of tumor were all observed every 2-3 days. After 5 weeks, when the mice were going to be dead, the organs and tumor tissues were thentaken out for volumes measurement and other pathological detections by HE stain. Orbital blood sampling was also performed for the immunofixation electrophoresis （IFE）. Results It was for 2-3 weeks after bearing cancer when the number of subcutaneous nodules began to reach a peak, and the nodule had the maximum volume after bearing cancer for 5-6 weeks. The death peak time of nude mouse was at the 7th week, and its median survival time （MST） was 52 days. The tumor tissues were proved as plasmocytes by HE stain for subcutaneous nodules. Inflammatory damage in liver was observed. Human monoclonal immunoglobulin could not be detected in the blood sample. Conclusion It is a successful modeling method to establish the nude mice model of human MM by injecting subcutaneously 2× 10^7 RPMI8226 cells after exposed to 3 Gy X-ray for 24 hours. Besides the merits of convenience and easy technique, X-ray exposure will benefit the tumor growth rate, which can lower the immunity of nude mice. It is difficult to detect any human monoclonal immunoglobulin in blood during the tumor growth. The inflammatory injuries in liver could be used as one of the indexes of the therapeutic effect of anticancer drugs in the future.