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

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.染色体众数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100.0%,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、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是首次成功建立的人直肠原发性恶性淋巴瘤裸鼠原位移植模型,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治疗提供了理想动物模型平台.

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

AIM:To establish an orthotopic transplantation model of human primary rectal lymphoma in nude mice and investigate its biologic features. METHODS:Histologically fresh lymphoma tis- sues from a case of primary rectal lymphoma during operation were transplanted into rectal mucosa of nude mice.Tumorgenecity,invasion, metastasis,morphology(light microscopy,elec- tron microscopy and immunohistochemistry), karyotype analysis and DNA content of ortho- topically transplanted tumors were studied. RESULTS:According to the new WHO classi- fication of malignant lymphoma,an orthotopic transplantation model of human primary rectal lymphoma(non-Hodgkin's,B cell,high ma- lignancy)in nude mice(HRBL-0305)was suc- cessfully established.Immunohistochemistry showed that 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.A total of 187 nude mice were used for transplantation.Both the growth rate of transplantation and resuscitation rate of liquid nitrogen cryopreservation were 100%.Liver metastasis rate was 45.4%;metasta- sis rates of both lymph node and celiac planting were 38.0%.Transplanted human primary rectal lymphoma could autonomously and invasively grow in the rectum of nude mice,and further developed hematogenous lymph node and celiac planting metastases.Orthotopically transplanted tumors were consistent with the original human tumor in histopathological and ultrastructural features,DNA content and chromosomal karyo- type. CONCLUSION:An orthotopic transplantation model of human primary rectal lymphoma is successfully established in nude mice for the first time,which can completely repeat the natu- ral clinicopathologic course of human primary rectal lymphoma,and metastasis pattern of the model is similar to that of clinical patients. HRBL-0305 provides an ideal animal model plat- form for the researches on biologic characteris- tics and experimental therapy of human primary rectal lymphoma.