人肝恶性淋巴瘤裸小鼠原位和皮下移植模型的建立及生物学特性的研究

Establishment and biological characteristics of subcutaneously and orthotopically transplanted models of human primary malignant lymphoma of the liver in nude mice

目的 探讨肝恶性淋巴瘤发病机制，为实验治疗提供工具。方法 采用人肝恶性淋巴瘤术中新鲜组织块分别植入裸小鼠肝实质内和肩胛间皮下，观察原位移植和皮下移植成瘤率，移植瘤的侵袭、转移；并进行形态学（光镜、电镜、免疫组织化学）、血清学[甲胎蛋白（AFP）、乙型肝炎病毒表面抗原（HBsAg）、乳酸脱氢酶（LDH）]榆测，染色体核型和流式细胞分析。结果在裸小鼠体内建成了同一人体瘤源人肝原发性恶性淋巴瘤裸鼠原位移植模型（HLBL-0102）和皮下移植模型（HLBL-0103）。移植瘤的病理组织学为肝非霍奇金B细胞性恶性淋巴瘤。免疫组织化学显示：CD19、CD20、CD45RO、CD79a阳性，CD3、CD7阴性。血清学AFP阴性，HBsAg阳性，LDH 1267．5U／L。染色体众数范围55～59条。移植瘤细胞DI值1．57～1．61，均为异倍体。HLBL-0102和HLBL-0103分别传至37代和31代，共移植裸鼠383只。肿瘤的移植生长率和液氮冻存复苏成活率均为100％。HLBL-0102移植瘤在裸鼠肝内自主侵袭性生长，瘤细胞侵入并破坏临近肝组织和门脉区内胆管及静脉，无其他组织、器官侵犯及远处淋巴结被累及。HLBL-0103在裸鼠皮下呈结节状局部生长。结论 HLBL-0102和HLBL-0103是首次建立成功的人肝恶性淋巴瘤裸鼠移植模型，完整地模拟了人肝恶性淋巴瘤患者的临床过程，为研究肝恶性淋巴瘤发病机制和实验治疗提供了理想的动物模型。

Objective To provide an experimental tool for exploring pathogenesis and experimental treatments of primary lymphoma of the liver. Methods Histologically intact lymphoma tissues from patients with primary lymphoma of the liver were transplanted into liver parenchyma and inter scapular subcutis of nude mice. Tumorgenecity, invasion, metastasis, morphological characteristics, karyotype analysis and DNA content of orthotopically transplanted tumors were investigated and serological test （AFP, HBsAg and LDH） were performed. Results Orthotopically and subcutaneously transplanted models of human primary malignant lymphoma of the liver in nude mice （named HLBL 0102 and HLBL-0103, respectively） were successfully established. Histopathology examination of transplanted tumors showed primary lymphoma of the liver （non Hodgkin＇s, B cell）. CD19, CD20, CD45RO and CD79a were positive, while CD3 and CD7 were negative. Serological test indicated that AFP was negative and HBsAg was positive. The number of chromosome was between 55 and 59. DNA index （DI） was 1.57— 1.61 （i. e. heteroploid）. HLBL 0102 and HLBL-0103 had been passaged for 37 and 31 generations. 383 nude mice were used for transplantation. The growth rate and resusci ration rate of liquid nitrogen eryopreservation of transplanted tumors were both 100%. HLBL-0102 grew autonomically and invasively in the liver of nude mice and damaged adjacent liver tissues, bile ducts and veins of portal arca. There was no involvement of other tissues and organs or distal lymph nodes. HLBL-0103 locally and nodously grew in subcutaneous tissue of nude mice. Conclusions Transplanted model of human primary malignant iymphoma of the liver in nude mice is successfully established, h can completely simulate the natural clinical process of primary iymphoma of the liver and provide an ideal animal model for researches on the pathogenesis and experimental therapies of primary lymphoma of the liver.