Establishment of a highly-metastatic model of human primary melanoma of the small intestine orthotopically transplanted in the small intestine of nude mice / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To provide an useful animal model for exploring metastatic biology and anti-metastatic therapy of primary malignant melanoma of the small intestine.</p><p><b>METHODS</b>A 49-year old male patient with malignant melanoma was treated by surgery, and the primary tumor in the small intestine and a metastatic tumor in the liver were removed. The diagnosis of malignant melanoma was confirmed by histopathology. Fresh melanoma tissue fragments taken from the primary intestinal tumor and hepatic metastatic tumor were orthotopically implanted into the mucosal layer of small intestine in nude mice, respectively. The tumor growth rate, invasion and metastasis of the transplanted tumors were observed. Light and electron microscopy, immunophenotype analysis, flow cytometry and karyotype analysis were carried out.</p><p><b>RESULTS</b>Fragments of the primary and liver metastatic malignant melanoma were successfully implanted in nude mice. After continuous passages in nude mice, an highly-metastatic model of human primary malignant melanoma of the small intestine (from the primary lesion) in nude mice (termed HSIM-0602) and a liver metastatic model of human primary malignant melanoma of the small intestine (originally from the liver metastatic lesion) in nude mice (termed HSIM-0603) were successfully established. Histological examination of the transplanted tumors revealed a high-grade melanoma of the small intestine. Immunohistochemical stainings of S-100 protein and HMB45 were positive. Many scattered melanosomes and melanin complex were seen in the cytoplasm of tumor cells. Chromosomal modal number was between 55 and 59. DNA index (DI) was 1.59 - 1.71, representing a heteroploid. The HSIM-0602 and HSIM-0603 tumor models had been maintained for 21 and 23 passages in nude mice, respectively. 227 nude mice were used for transplantation. Both the growth rate after transplantation and resuscitation rate from liquid nitrogen cryopreservation were 100%. The HSIM-0602 model exhibited 84.8% lung metastasis, 65.7% liver metastasis and 63.8% lymph node metastasis. However, HSIM-0603 displayed 100% liver metastasis, 46.7% lung metastasis and 71.3% lymph node metastasis. The transplanted tumors actively and invasively grew in the small intestine of nude mice and showed hematogenous and lymphatic metastases.</p><p><b>CONCLUSION</b>To our knowledge it is the first time that two strains of spontaneous highly-metastatic nude-mouse model of human primary malignant melanoma of the small intestine have been successfully established in our department. The models are very closely mimic the natural clinicopathologic course of primary small intestinal melanoma in humans and provide ideal animal models for the researches on metastasis biology and anti-metastatic experimental therapy of malignant melanoma of the small intestine.</p>

Establishment of a nude mouse model of orthotopically transplanted human primary malignant lymphoma of the liver / 中华肿瘤杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

Establishment and characteristics of orthotopically transplanted model of human primary malignant small intestinal lymphoma in nude mice / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To establish orthotopically transplanted model of human malignant small intestinal lymphoma in nude mice and analyze their biologic characteristics.</p><p><b>METHODS</b>Small intestinal lymphoma tissues from 5 patients were transplanted into intestinal mucosa of nude mice. Tumorgenecity, invasion and metastasis of the transplanted tumors were observed by morphological analyses (light microscopy, electron microscopy and immunohistochemistry), karyotyping and DNA quantitative assay.</p><p><b>RESULTS</b>Tumor tissues from 3 lymphoma patients were successfully transplanted. According to the World Health Organization classification, the three models were classified into non-Hodgkin's lymphoma (B cell) of human small intestine (HSIL-1), high metastasis of non-Hodgkin's lymphoma (B cell) of human small intestine (HSIL-2) and non-Hodgkin's lymphoma (T cell) of human small intestine (HSIL-3), respectively. Immunohistochemistry showed that CD19, CD20, CD22, CD40, CD45 and CD72 were positive in HSIL-1 and HSIL-2, whereas CD3, CD7 and CD45RO were positive in HSIL-3. The karyotypes of the transplanted tumors were all hypotriploid with modal numbers from 55 to 69 and the DNA index (DI) was 1.46 approximately 1.71. The three models had been passaged for 32, 27 and 21 generations respectively in 433 nude mice. The growth rate, resuscitation rate of the liquid nitrogen preserved tumor cells and spontaneous metastasis rate upon transplantation were all 100%. We observed an invasive growth of the transplanted tumors in small intestine, which resulted in disrupting of the intestinal wall, hematogenous metastasis, lymph node metastasis and seeding metastasis. The features of the transplanted tumors were similar to the original tumors in histopathology, ultrastructure, DNA content and karyotype.</p><p><b>CONCLUSION</b>Three strains of orthotopically transplanted model of human primary malignant small intestinal lymphoma in nude mice were successfully developed. The result of research will provide ideal animal models for further studies on mechanism of tumorigenesis, invasion and metastasis of malignant small intestinal lymphoma and experimental therapy.</p>