骨与软组织肉瘤PDX模型的建立及应用研究

Study of patient-derived xenograft model of bone and soft tissue sarcoma and its application

目的建立恶性骨与软组织肉瘤患者来源移植瘤模型（patient-derived xenograft，PDX）模型并统计其建模的成功率，观察PDX化疗反应与临床的符合率，以为筛选敏感二、三线药物提供理论基础。方法收集自2015年1月至2016年5月共31例骨与软组织肿瘤患者，男12例，女19例；年龄为9~67岁，平均（28.5±19.9）岁。手术当天取材，取患者手术中新鲜的活性组织，无菌条件下转移到实验室后，切至约2 mm3大小，接种于采用穿刺套管针接种于T淋巴细胞缺陷小鼠（BAL B/C裸鼠）或T、B淋巴细胞双缺小鼠（SCID小鼠）背部皮下，待小鼠背部肿瘤长至1~2 cm直径时取出肿瘤组织并传代，同时保存P0组织；若小鼠背部肿瘤3个月无生长则视为建模失败；分别接种化疗前和化疗后的病例，比较化疗对PDX建模成功率的影响，同时观察不同病理类型病例的建模成功率，观察PDX建模成功率与患者预后的关系；药物试验：将成瘤的PDX小鼠分组，每组3只，不同组给予不同的化疗药物，分别按照化疗药物特性给药，通过T/C计算抑制率，比较得出PDX模型敏感药物及不敏感药物，并与对应的患者用药情况进行比较。结果本组共接种骨与软组织肉瘤31例，总体建模成功率45.2%。按病理亚型分别为：骨肉瘤24例，建模成功率37%；平滑肌肉瘤2例，成功率100%；软骨肉瘤2例，成功率50%；尤文肉瘤1例成功；纤维肉瘤1例及滑膜肉瘤1例均未成功。化疗后建模成功率33%（4/12），未化疗建模成功率53%（10/19）。PDX建模较容易的患者往往提示预后不佳，PDX模型对化疗的敏感性与临床实际疗效具有较好的相符合性。结论骨与软组织肉瘤的PDX成功率在30%~40%，与肿瘤病理类型、化疗有关，PDX与临床病例具有较好的相似性，有望用于患者个体化药物筛选。

Objective Create patient-derived xenografl （PDX） model of bone and soft tissue sarcoma, and analyze the success rate of PDX model, observe the effects of chemotherapy on PDX models and its coincidence, and provide a theoretical ba- sis for screening sensitive second and third line drugs. Methods Collected 31 cases of bone and soft tissue sarcoma from January 2015 to May 2016, which included 12 male and 19 female, with an average age of （28.5±19.9） y. The tumor tissue was obtained the day of operation, and it was cut into 2 mm^3 pieces and injected into the flank of BAL B/C nude mice or SCID mice. Tumor was passaged when the diameter reached 1-2 cm and the P0 tissue was froze. If there was no obvious tumor mass grows out for 3 months, the model creation will be stopped. We inoculated the mice with patients sample with or without chemotherapy, observed the effect of chemotherapy on the success rate of PDX modeling and the success rate of modeling of different pathological types, and also observed the relationship between the success rate of PDX modeling and the prognosis of patients. For the drug sensitivity test, 3 mice was used in each group, and chemotherapy was given, T/C was used to evaluate the inhibition ratio after drug treat- ment. Results 31 PDX models were inoculated. The total sueeess rate is 45.2%. Pathology of the PDX models and their success rates： 24 osteosareoma models, success rate is 37%; 2 leiomyosareoma models, success rate is 100%; 2 ehondrosarcoma models, success rate is 50%; 1 Ewing sareoma model sueeessed; 1 fibrosareoma model and 1 synovial sarcoma model, were not sueeessed. Post chemotherapy model success rate is 33% （4/12）, compared with 53%（10/19） of model success rate that without chemotherapy. And there is relationship between sueeess rate of PDX model creation and patient outcome. The faster the PDX model creation, the worse the outcome, The drug sensitivity of PDX model coincides the elinical situation. Conclusion The success rate of creating PDX model of bone and soft tissue sarcoma is around 30%-40%, and it is related to the pathology and whether got ehemotherapy or not, PDX models coincide sarcomas clinical situation, and it is hopefully to use PDX model in selecting personalized drugs.