人原发性肝恶性淋巴瘤裸小鼠原位移植模型的建立

目的建立人肝原发性恶性淋巴瘤裸小鼠原位移植模型,为探讨肝恶性淋巴瘤发病机制和实验治疗提供工具。方法采用人肝原发性恶性淋巴瘤术中新鲜瘤组织块植入裸小鼠肝实质内,观察原位移植成瘤率和移植瘤的侵袭、转移,并进行形态学（光镜、电镜、免疫组织化学）、血清学、染色体核型和流式细胞分析。结果在裸小鼠体内建成了一株人肝原发性恶性淋巴瘤原位移植模型（HLBL-0102）。移植瘤的组织病理学为原发性肝恶性淋巴瘤（非霍奇金B细胞性）,免疫组织化学显示,CD19、CD20、CD45和CD79a阳性,CD3、CD7阴性,甲胎蛋白（AFP）阴性,乙型肝炎病毒表面抗原（HbsAg）阳性,乳酸脱氢酶（LDH）1267．5U／L。染色体众数范嗣55—59条;移植瘤细胞DNA指数值1．57～1．61,均为异倍体。HLBL-0102住裸鼠体内生长3年,已经传至37代,共移植裸鼠283只,其肿瘤的移植生长率和液氮冻存复苏成活牢均为100％。人原发性肝恶性淋巴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累及。结论HLBL-0102是首次建苞成功的人原发性肝恶性淋巴瘤裸鼠原位移植模型,完整地模拟了人原发性肝恶性淋巴瘤患者的自然临眯病理过程,为研究原发性肝恶件淋巴瘤生物学和实验治疗提供了理想的动物模型。

Establishment of a nude mouse model of orthotopically transplanted human primary malignant lymphoma of the liver

OBJECTIVE: To establish a nude mouse model of orthotopically transplanted human primary malignant lymphoma of the liver, and to serve researches on pathogenesis and experimental treatment of this disease. METHODS: Small pieces of lymphoma tissues freshly taken from patients with primary lymphoma of the liver were orthotopically transplanted into the liver parenchyma in nude mice. Tumorgenicity, invasion, metastasis, and morphological characteristics were examined by light and electron microscopy and immunohistochemistry. AFP, HBsAg and LDH were assayed by serological test. Karyotype analysis and DNA content of orthotopically transplanted tumors were also performed. RESULTS: A nude mouse model of orthotopically transplanted human primary malignant lymphoma of the liver was successfully established and named HLBL-0102. The tumor was confirmed as primary lymphoma of the liver (non-Hodgkin's lymphoma, B cell) by histopathology. Immunohistochemistry showed positive expression of CD19, CD20, CD45 and CD79a, but negative of CD3 and CD7. Serological test indicated that AFP was negative, HBsAg positive and the concentration of LDH was 1267.5 U/L. The number of chromosomes was between 55 and 59. DNA index (DI) was 1.57 approximately 1.61 (i.e. heteroploid). So far, the strain HLBL-0102 has grown for 3 years and been passaged for 37 generations in nude mice. Totally 283 nude mice were used for transplantation and the successful rate was 100%. Both the growth rate and resuscitation rate of liquid nitrogen cryo-preserved transplanted tumors were 100%. The transplanted tumors grew intensely and invasively in the liver of nude mice and damaged adjacent liver tissues, bile ducts and portal vein areas. No involvement of other tissues and organs and distal lymph nodes was observed. CONCLUSION: To our knowledge it is the first report of successfully established nude mouse model of orthotopically transplanted human primary malignant lymphoma of the liver. The HLBL-0102 model simulate very well the natural process of human primary lymphoma of the liver and provides an ideal animal model for researches on the biology and therapies of this malignancy.