Establishment of a nude mouse model of highly metastatic gastric lymphoma constructed with orthotopic transplantation of surgical specimen / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To construct a mouse model of highly metastatic gastric lymphoma with orthotopic transplantation of human primary gastric lymphoma specimen.</p><p><b>METHODS</b>A fresh surgical specimen of primary gastric lymphoma was obtained intraoperatively and implanted into the submucosa of stomach in nude mice. Tumor formation, invasion, metastasis, morphological characteristics under light microscopy and electron microscopy, immunohistochemistry,and the karyotype of orthotopically transplanted tumor cells were studied.</p><p><b>RESULTS</b>An orthotopic highly metastatic model of human primary gastric lymphoma in nude mice(HGBL-0305) was successfully established. Histopathology of transplanted tumors showed primary gastric diffuse large B cell lymphoma. CD19, CD20, CD22 and CD79alpha were positive, while CD3 and CD7 were negative. The number of chromosome ranged from 56 to 69. DNA index(DI) was 1.47+/-0.12(i.e. heteroploid). Until now, HGBL-0305 model has been maintained for 45 generations by orthotopic passage for almost 4 years in nude mice. A total of 156 nude mice were used for transplantation. The growth rate and resuscitation rate of liquid nitrogen cryopreservation of transplanted tumor cells were both 100%. The autonomic growth of the transplanted tumor cells invaded and destructed all the layers of the nude mice stomach. The metastasis rates of liver, spleen, lymph node, and peritoneal seeding were 69.5%, 55.6%, 45.7%, and 30.5%, respectively.</p><p><b>CONCLUSIONS</b>An orthotopic highly metastatic model of human primary gastric lymphoma in nude mice is successfully established. HGBL-0305 model may simulate the natural course of primary gastric lymphoma in human and provides an ideal animal model for studies on pathogenesis, metastasis biology and anti-metastatic therapies of primary gastric lymphoma.</p>

A liver-metastatic model of human primary gastric lymphoma in nude mice orthotopically constructed by using histologically intact patient specimens / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>In recent years, incidence and mortality of lymphoma are markedly increasing worldwide. However, the pathogenesis and mechanism of invasion and metastasis for lymphoma are not yet fully clarified. It is mainly due to the lack of ideal animal models, which can precisely simulate the invasion and metastasis of lymphoma in the human body. So, it is very necessary to establish a highly metastatic nude mouse model of human lymphoma. This study developed a liver-metastatic model of primary gastric lymphoma in nude mice by using orthotopic surgical implantation of histologically intact patient specimens into the corresponding organs of the recipient small animals.</p><p><b>METHODS</b>A histologically intact fragment of liver metastasis derived from a surgical specimen of a patient with primary gastric lymphoma was implanted into the submucosa of the stomach in nude mice. Tumorigenicity, invasion, metastasis, morphologic characteristics (via light microscopy, electron microscopy, and immunohistochemistry), karyotype analysis, and DNA content of the orthotopically transplanted tumors were studied.</p><p><b>RESULTS</b>An orthotopic liver metastatic model of human primary gastric lymphoma in nude mice (termed HGBL-0304) was successfully established. The histopathology of the transplanted tumors showed primary gastric diffuse large B-cell lymphoma. CD19, CD20, CD22, and CD79alpha were positive, but CD3 and CD7 were negative. The serum level of lactate dehydrogenase (LDH) was elevated [(1010.56+/-200.85) U/L]. The number of chromosomes ranged from 75 to 89. The DNA index (DI) was 1.45+/-0.25 (that is, heteroploid). So far, the HGBL-0304 model has been passed on for 45 generations of nude mice. A total of 263 nude mice were used for the transplantation. Both the growth and resuscitation rates of liquid nitrogen cryopreservation of the transplanted tumors were 100%. The transplanted tumors autonomically invasively grew and damaged a whole layer in the stomach of nude mice. The metastasis rates of liver, spleen, lymph node, and peritoneal seeding were 100%, 94.3%, 62.6%, and 43.5%, respectively.</p><p><b>CONCLUSIONS</b>The study successfully establishes an orthotopic liver metastatic model of human primary gastric lymphoma in nude mice. The HGBL-0304 model can completely simulate the natural clinical process of primary gastric lymphoma and provides an ideal animal model for the research on the biology of metastasis and antimetastatic experimental therapies of primary gastric lymphoma.</p>

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 nude mice liver metastatic model of human primary malignant melanoma of the small intestine / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To provide ideal animal models for exploring pathogenesis and experimental therapy of primary malignant melanoma of the small intestine.</p><p><b>METHODS</b>The histologically intact primary and liver metastatic fragments derived from surgical specimens of one patient with metastatic malignant melanoma of the small intestine were orthotopically implanted in the small intestinal mucous layer of nude mice. The take rate, invasion and liver metastasis were observed. Morphology (light microscopy, electron microscopy), immunophenotype analysis, flow cytometry and karyotype analysis were applied for the original human tumors and the transplanted tumors.</p><p><b>RESULTS</b>The primary and liver metastatic fragments of malignant melanoma of the small intestine were successfully implanted in nude mice. After continuous passages in nude mice,an orthotopic model of human primary malignant melanoma of the small intestine(from the primary focus)in nude mice (termed HSIM-0501) and a liver metastatic model of human primary malignant melanoma of the small intestine (from the liver metastatic focus) in nude mice (termed HSIM-0502) were established. Histological examination of transplanted tumors revealed high-grade melanoma. S-100 protein and HMB45 were positive. Massive melanin granules and melanin complex were seen in cytoplasm of tumor cells.Chromosomal modal number was between 55 and 59. DNA index (DI) was 1.49-1.61, representing heteroploid. HSIM-0501 and HSIM-0502 were maintained for 25 and 27 passages in nude mice respectively. Three hundred and seventeen nude mice were used for transplantation. Both the take rate after transplantation and resuscitation rate of liquid nitrogen cryopreservation were 100%. HSIM-0501 exhibited 46.2% liver metastasis and 36.7% lymph node metastases. In HSIM-0502, both liver and lymph node metastases were 100%.The transplanted tumors autonomically and invasively grew in the small intestines of nude mice and hematogenous metastasis, lymph node metastasis and celiac planting metastasis occurred.</p><p><b>CONCLUSION</b>Two nude mice liver metastatic models of human primary malignant melanoma of the small intestine are successfully established, which provide ideal animal models for the research of pathogenesis,metastasis biology and anti-metastatic experimental therapy of primary malignant melanoma of the small intestine.</p>

AF172993 sequence of Plunc in GenBank database is not the complete CDS / 南方医科大学学报

<p><b>OBJECTIVE</b>To determine AF172993 sequence is either the complete CDS or a transcript variant.</p><p><b>METHODS</b>RT-PCR was used to amplify the CDS sequence of Plunc, which was subsequently cloned into the pEGFP-N1 eukaryotic expression vector. After bi-directional sequence analysis, the sequence obtained was blasted against the AF172993 sequence, nr database and human genome database.</p><p><b>RESULTS</b>In CDS of the new cloned sequence, the 658 base A in the AF172993 sequence was replaced by C, and the corresponding genetic code was also converted from AAG to CAG, leading to the alteration of the amino acid Gln to Lys. In addition, the base C at the 658 position of the CDS showed perfect match with the base C at 2094188 position in human chromosome 20.</p><p><b>CONCLUSION</b>The base A at the 658 position of AF172993 sequence of Plunc is a mutation site, which alters the coding of the amino acid. AF172993 sequence is actually a transcript variant of Plunc, and the annotation to AF172993 in GenBank database is not correct and need to be revised.</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 of liver metastasis model of human primary colonic lymphoma by orthotopic transplantation / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To provide an ideal animal model for exploring pathogenesis and experimental treatment of primary colonic lymphoma.</p><p><b>METHODS</b>Primary colonic and liver metastatic lymphoma tissues were obtained from the surgical specimens,and transplanted into colonic mucosa of nude mice respectively. The tumorigenesis, invasion, metastasis and morphology of the transplanted tumor were observed. Karyotype was analyzed and DNA content was measured.</p><p><b>RESULTS</b>According to the new WHO classification of malignant lymphoma, two high metastatic models (HCBL-0303 from primary lymphoma and HCBL-0304 from live metastatic lesion) of human primary colonic non-Hodgkin's B cell lymphoma in nude mice were established successfully by orthotopic transplantation. Pathological examination showed poorly differentiated non-Hodgkin's B cell lymphoma of the transplanted tumors, and immunohistochemical staining showed positive expressions of CD19, CD20 and CD22, and negative expressions of CD3 and CD7. The number of chromosome ranged from 55 to 59, and DNA index (DI) was 1.59 - 1.71 (i.e. heteroploid). In HCBL-0303,liver metastasis rate was 63.7% and lymph node metastasis rate was 56.4%. However, in HCBL-0304, both metastasis rates of liver and lymph node were 100%. The transplanted tumors grew autonomously and invasively in nude mice, and further developed hematogenous, lymphatic metastasis and intraperitoneal seeding.</p><p><b>CONCLUSIONS</b>HCBL-0303 and HCBL-0304 are the first established high metastatic models of primary colonic lymphoma, and can be applied to the research on pathogenesis, invasion,metastasis and experimental therapy of human primary colonic lymphoma.</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>