人原发性直肠恶性淋巴瘤裸小鼠原位移植模型的建立及其生物学特性

Establishment of orthotopic transplantation model of human primary rectal lymphoma in nude mice and investigation of its biologic characteristics

目的 建立人原发性直肠恶性淋巴瘤裸小鼠原位移植模型，探讨其生物学特性。方法 采用人直肠原发性恶性淋巴瘤切除术中的新鲜瘤组织块植入裸鼠的直肠黏膜层内，观察原位移植的成瘤率、移植瘤的侵袭和转移率。进行形态学（光镜、电镜）、免疫组织化学、染色体核型、流式细胞分析。结果 依据WHO新的分类标准，建成1株人直肠原发性（非霍奇金B细胞性）恶性淋巴瘤裸鼠原位移植模型HRBL-0305。移植瘤组织病理学为（非霍奇金B细胞性）高度恶性淋巴瘤，免疫组织化学示CD19、CD20、CD22、CD45阳性，CD3、CD7阴性，染色体56～69条，流式细胞DI值为1．57～1．61，均为异倍体。HRBL-0305已传至31代，共移植裸鼠187只。其肿瘤移植生长率和液氮冻存复苏成活率均为100％。肝转移率为45．4％，淋巴结和腹腔种植转移率均为38．0％，移植瘤在裸鼠的直肠内自主侵袭性生长，发生血液转移、淋巴转移和腹腔内种植性转移。移植瘤组织病理学、超微结构的观察、流式细胞DNA含量测定及染色体核型的分析，表明与人源直肠恶性淋巴瘤细胞相一致。结论 HRBL-0305是首次建立成功的人直肠原发性恶性淋巴瘤裸鼠原位移植模型。该模型完整地重现了人直肠原发性恶性淋巴瘤的自然临床病理过程，且转移模式与临床患者相似。为研究直肠恶性淋巴瘤的生物学特性和实验治疗提供了理想动物模型平台。

Objective To establish an orthotopic transplantation model of human primary rectal lymphoma in nude mice and investigate its biologic features. Methods Histologically fresh lymphoma tissues from a case of primary rectal lymphoma were transplanted into rectal mucosa of nude mice. Tumorgenecity, invasion, metastasis, morphology （light microscopy, electron microscopy and immunohistochemistry） and DNA content of orthotopically transplanted tumors were studied and karyotype analyzed. Results According to the new WHO classification of malignant lymphoma, an orthotopic transplantation model of human primary rectal lymphoma （non-Hodgkin＇s, B cell） in nude mice （HRBL-0305） was successfully established. Histopathological examination of transplanted tumors showed non-Hodgkin＇s B cell lymphoma. CD19, CD20, CD22 and CD45 were positive, but CD3 and CD7 were negative. The number of chromosome was between 56 and 69. DNA index （DI） was 1. 57-1. 61 （i. e. heteroploid）. So far, HRBL-0305 has been passaged for 31 generations in nude mice. 187 nude mice were used for transplantation. Both the growth rate after transplantation and resuscitation rate of cells from liquid nitrogen cryopreservation were 100%. Liver metastasis rate was 45.4% metastasis rates of both lymph node and celiac planting were 38.0%. Transplanted human primary rectal lymphoma could autonomously and invasively grew in the rectum of nude mice, and hematogenous, lymph node and celiac planting metastases occurred. Orthotopically transplanted tumors were consistent with the original human tumor in histopathological and ultrastructural features, DNA content and chromosomal karyotype. Conclusions The study successfully establishes an orthotopic transplantation model of human primary rectal lymphoma in nude mice. In this model, the clinico-pathologic course of human primary rectal lymphoma and metastasis pattern of the model were similar to those of the clinical patients.