Growth inhibition of human melanoma tumor xenografts in athymic nude mice by swainsonine

Swainsonine, an inhibitor of alpha-mannosidases, has been shown to block experimental metastasis of B16F10 melanoma and MDAY-D2 lymphoid tumor cells in syngeneic mice. In this report we demonstrate that swainsonine also reduces the growth rate of human melanoma cells in vitro and in vivo. Graded doses of swainsonine were administered either orally or via implanted Alzet miniosmotic pumps to athymic nude mice bearing subcutaneously implanted human MeWo melanoma cells. Swainsonine at 10 micrograms/ml in the drinking water or 0.5 mg/kg/day administered by miniosmotic pump reduced the growth rate of the MeWo tumors by approximately 50% and inhibited the expression of complex-type oligosaccharides in tumors and host intestine by only 10-20%. Swainsonine doses of 4 mg/kg/day reduced expression of complex-type oligosaccharides by 85% in vivo but afforded no additional inhibitory effect. A glycosylation mutant of MeWo called 3S5 has a defect in the synthesis of complex-type asparagine-linked oligosaccharides resulting in incomplete processing similar to that observed in swainsonine-treated MeWo tumor cells. Swainsonine did not inhibit the proliferation of 3S5 cells in vitro nor the growth of 3S5 tumors in nude mice. The results suggest that expression of highly branched complex-type oligosaccharides commonly associated with the malignant phenotype may provide the tumor cells with a growth advantage.     

Detection of metastatic tumors in nude mice: brief communication.

Homozygous nude (nu/nu) mice were inoculated ip with either highly malignant human bladder transitional cell carcinoma or human prostate adenocarcinoma. These animals were subsequently given injections of normal human T-lymphocytes to restore the known T-lymphocyte deficiency present in homozygous nude mice. Metastatic spread of the prostate and bladder carcinomas was evident in mice given human T-lymphocytes. Although tumor growth was observed at the sites of tumor inoculation, no tumor spread was observed in mice not receiving T-lymphocytes.     

Metastases of human tumor xenografts in nude mice

In the present study, using systematic microscopic examination, we tried to determine the true incidence of metastases in nude mice bearing a wide variety of human tumors. A total of 63 malignant tumors were successfully transplanted subcutaneously and 831 nude mice bearing tumors were examined. It appeared that 17 of the 63 tumors (26.9%) retained their metastatic ability in nude mice. Most of these tumors were adenocarcinomas (11/17 cases). Generally the metastatic deposits in the lungs and, to a lesser extent, in the lymph nodes were small and thus only detectable on microscopic examination. We also found a positive correlation between the presence of metastases and neoplastic infiltration of the lymphatic and/or blood vessels around the subcutaneous tumors. Metastatic human tumors, including neoplastic cells from effusion, exhibited higher metastatic ability than primary tumors (p less than 0.005). However, the expression of this metastatic potential depends on several factors including tumor volume, survival time after inoculation and murine hepatitis infection. Thus, animals with metastases bore larger tumors (9.56 cm3) than those without metastasis (6.35 cm3; p less than 0.0001). Moreover, survival time after inoculation was longer in mice with metastases (104 days) than in mice without metastases (81 days; p less than 0.0001). A negative influence of viral hepatitis on the incidence of metastases was observed. This may simply be related to the shortened life span of the animals. Death due to this infection may precede the expression of the metastatic potential.     

Growth of human bronchial carcinomas in nude mice

Two hundred and thirteen lung tumours of primary site and 42 metastases were heterotransplanted into nude mice with an overall success rate of 44%. There were differences in success between the histological types. Squamous cell and adenocarcinoma had the highest success rate (51% and 43%, respectively) whereas large cell and small cell carcinoma had a lower success rate (38% for both). The average volume doubling times in the first passage in nude mice ranged from 8.2 in large cell carcinomas to 18.9 days in adenocarcinomas. In subsequent passages an increase in growth rate was found, the overall average doubling time falling from 14.5 days in the first passage to 7.1 days in the second passage. In a study with 171 non-small cell lung carcinomas (NSCLC), the growth data in nude mice were correlated with the clinical data of the corresponding patients. A relationship between the growth parameters in nude mice and prognosis of patients could not be found.     

Inhibition of human tumor growth by intraperitoneal immunotoxins in nude mice

Intracavitary administration of immunotoxins may play a role in the control of malignant effusions. Selection of immunotoxins for this form of therapy is based on their prior evaluation in preclinical studies. Monoclonal antibodies (mAb) 454A12 (antitransferrin receptor), and 260F9 are directed against antigens which are present on tumor cells in pleural and peritoneal effusions of patients with adenocarcinoma of the breast and ovary. In the present study, immunotoxins derived by conjugating these mAb to recombinant ricin A (rRA) were shown to be cytotoxic to human ovarian adenocarcinoma HEY cells in vitro and in vivo. In the in vitro assay 454A12-rRA and 260F9-rRA were 1000-fold and 10-fold, respectively, more cytotoxic than free rRa against HEY cells, and both immunotoxins were potentiated approximately 1000-fold by monensin. For in vivo studies HEY cells were injected i.p. into nude mice at a challenge dose (3 x 10(5) cells) which produced carcinomatosis with ascites, leading to death 30 days following injection. Administration of 454A12-rRA i.p. following the challenge dose resulted in a complete cure, whereas administration of 260 F9-rRA with monensin significantly prolonged survival. The greater cytotoxicity of 454A12-rRA than 260F9-rRA against HEY cells could be accounted for by the greater number of binding sites and higher internalization rate for 454A12-rRA and mAb 454A12 than 260F9-rRA and mAb 260F9, respectively. These results suggest a potential role for 454A12-rRA and 260F9-rRA plus monensin in the intracavitary therapy of malignant effusions associated with carcinoma of breast and ovary. In the case of 260F9-rRA, this represents the first preliminary indication of the suitability of this immunotoxin for intracavitary therapy of malignancies.     

Inhibition of the growth of human melanoma xenografts in nude mice by human tumor-specific cytotoxic T-cells

Melanoma-specific T-cells (CTLs) are specifically cytotoxic for autologous tumor, when assayed in vitro. To examine their effectiveness in vivo, we tested the ability of these human T-cells to inhibit growth of human melanoma xenografts by using a Winn assay. Nude mice receiving specific CTLs (n = 10) demonstrated a dramatic inhibition of tumor growth. All treated mice were tumor-free at day 50 and nine remained tumor-free at day 65, vs. control mice (n = 10) with average tumor volumes of 321 mm3 and 808 mm3, respectively. To control for the possibility that a non-specific response to the human T-cells could inhibit tumor growth, an additional group received allospecific CTLs. There was no inhibition of tumor growth in this group (n = 8), with the average tumor volume of 2,768 mm3 at day 40 vs. 1,882 mm3 in the control group (n = 10). We conclude that these tumor-specific CTLs can inhibit tumor growth in vivo and may prove useful in the adoptive immunotherapy of melanoma.     

Inhibition of the growth of human melanoma metastases in nude mice by melanoma-specific murine monoclonal antibody.

The administration of anti-melanoma murine monoclonal antibody (MAB) 16.C8 (IgG2a) to nude mice bearing established human melanoma lung or liver metastases resulted in a significant inhibition of tumour growth. A total dose of 2 mg of affinity purified 16.C8 caused complete inhibition of tumour growth in 89 and 100% of animals in the liver and lung model, respectively. In contrast, a significant tumour growth was found in most control animals which received an irrelevant IgG2a MAB or 2% human serum albumin in Hanks Balanced Salt Solution (HBSS). The MAB was most effective when treatment was started on day 1 or 4 following tumour inoculation. When the 16.C8 MAB treatment was delayed 7 or 14 days, 33 and 67% of 16.C8 treated animals, respectively, developed tumours. The MAB-mediated anti-tumour activity appeared to be dose dependent, and the effect of a suboptimal dose was potentiated by the concomitant administration of recombinant interleukin 2 (rIL-2). Recombinant IL-2 alone in a similar dose did not elicit comparable anti-tumour activity. Moreover, the MAB 16.C8 inhibited tumour growth in irradiated animals which may suggest the involvement of host-radioresistant cellular elements in the 16.C8 antibody-mediated anti-tumour activities in nude mice. These results suggest that MAB 16.C8 alone or combined with rIL-2 may prove useful in the immunotherapy of metastatic melanoma.     

Soluble intercellular adhesion molecule 1 is released by human melanoma cells and is associated with tumor growth in nude mice.

We have studied the cytokine regulation of cell surface and soluble intercellular adhesion molecule 1 (ICAM-1) expression on the human melanoma cell line A375M. Unstimulated cells express ICAM-1 on their cell surface but do not secrete significant levels of soluble ICAM-1. Interleukin 1, interleukin 6, tumor necrosis factor, and gamma-interferon all increased cell surface expression of ICAM-1. Tumor necrosis factor, interleukin 1, and gamma-interferon also caused the release of soluble ICAM-1. The serum of melanoma patients has been reported to contain elevated levels of soluble ICAM-1; however, the source of this ICAM-1 is unclear. The serum from nude mice bearing s.c. human melanoma tumors was found to contain soluble human ICAM-1. ICAM-1 levels showed a positive correlation with tumor weight. The release of ICAM-1 from melanoma tumors, in response to host-derived cytokines, may have relevance to immune recognition of the tumor.     

去势后人子宫内膜癌裸鼠移植瘤模型的建立

目的:研究建立去势后人子宫内膜癌裸鼠移植瘤模型的方法.方法: 将裸鼠在麻醉情况下切除双侧卵巢,随机分成3组,采用细胞悬液法和肿瘤组织块法建立人子宫内膜癌的裸鼠皮下移植瘤模型.结果: 细胞悬液法5×106 /0.2ml/只,瘤块法1.5mm×1.5mm×2mm和1.0mm×1.0mm×1.0mm,三种植瘤法成瘤率无统计学意义,但不同的植瘤法成瘤时间有差异;各组肿瘤体积大小不同,差异有统计学意义.结论: 选择适当的方法建立人子宫内膜癌去势裸鼠移植瘤模型是可行的,可为开展人子宫内膜癌的基础和临床研究提供理想的动物模型.     

Monitoring the therapy of human tumor xenografts in nude mice by the use of lactate dehydrogenase.

With the use of circulating human lactate dehydrogenase (LDH) as a tumor marker, growth and remission of human tumor lines SW480, HEp-2, and Clouser, implanted into female BALB/c athymic nude mice, were followed during therapy. Three types of therapy were used: X-radiation, cyclophosphamide, and diphtheria toxin. After therapy tumor sizes were measured with calipers and compared to changes in the levels of circulating human LDH. Changes in LDH levels paralleled changes in tumor size, but the enzyme fluctuations were more pronounced. Mice bearing intraperitoneally growing SW480 and HEp-2 tumors were effectively treated with diphtheria toxin, and the measurement of circulating LDH was examined as a parameter for gauging the effectiveness of chemotherapy on tumors that could not be visualized. Circulating human LDH can be used to detect intraperitoneal tumor growth and/or remission and to predict death of the animal due to the tumor.     

Exceptional lethality for nude mice of cells derived from a primary human melanoma

BRO human melanoma cells, obtained from a biopsy of a highly aggressive and malignant primary tumor, were grown as xenografts in nude mice and in cell culture. These cells were exceptionally tumorigenic and malignant for nude mice. NIH-II nude mice survived 11.0 +/- 0.4 (S.E.) and 14.1 +/- 0.4 days after i.p. inoculation of 10(7) or 10(6) BRO cells, respectively, and lethal tumors developed in all mice inoculated i.p. with only 10(3) cells. The doubling time (2.3 days) of the volume of tumors formed after s.c. inoculation was comparable to the doubling time of these cells in culture. After i.p. or s.c. inoculation, BRO cells metastasized to the diaphragm and lungs, causing respiratory failure in most of the host mice. The original tumor and the cell line derived from it had undifferentiated structures with prominent nuclei and very large nucleoli. Karyotype abnormalities included a gigantic A group chromosome, a large D group chromosome, and an unusual double centromere chromosome not found typically in human melanoma cells. Due to the short and reproducible survival times of nude mice inoculated i.p. with BRO cells, this model system may be useful for rapidly determining the effects of experimental treatment on the survival of hosts bearing human tumor cells.     

Interleukin 1-induced augmentation of experimental metastases from a human melanoma in nude mice

This study has examined the effect of the cytokine interleukin 1 (IL-1) on metastasis formation by the human melanoma A375M in nude mice. We have found that human recombinant IL-1 beta (a single injection greater than 0.01 micrograms per mouse i.v. given before tumor cells) induced an augmentation of experimental lung metastases from the A375M tumor cells in nude mice. This effect was rapidly induced and reversible within 24 h after IL-1 injection. A similar effect was induced by human recombinant IL-1 alpha and human recombinant tumor necrosis factor, but not by human recombinant interleukin 6. 5-[125I]odo-2'-deoxyuridine-radiolabeled A375M tumor cells injected i.v. remained at a higher level in the lungs of nude mice receiving IL-1 than in control mice. In addition, IL-1 injected 1 h, but not 24 h, after tumor cells enhanced lung colonization as well, thus suggesting an effect of IL-1 on the vascular transit of tumor cells. These findings may explain the observation of enhanced secondary localization of tumor cells at inflammatory sites and suggest that modulation of secondary spread should be carefully considered when assessing the ability of this cytokine to complement cytoreductive therapies.     

First heterotransplantation of a human carcinoid tumor into nude mice.

The first successful heterotransplantation of a human carcinoid tumor into nude mice is reported. CSH, a voluminous hepatic metastasis of a primary bronchial carcinoid tumor (CSB) was resected and transplanted into three irradiated nude (Swiss-nu/nu) mice both by subcutaneous (SC) and intramuscular (IM) routes; the success rate was five of six. Heterotransplanted tumors took 4 to 5 months to appear in the mice and 1 month to attain a width of 0.5 cm. Both human and mouse tumors (named CSH-SC and CSH-IM) were studied by light and electron microscopy. They were Grimelius-positive, neuron-specific enolase-positive, and bombesin-negative by immunocytochemistry. Furthermore, CSH-SC cells presented characteristic (pear-shaped, rod-shaped, or tadpole-shaped) neurosecretory granules. Although CSB and CSH were slightly serotonin positive by immunocytochemistry, only a few serotonin-positive cells were found in CSH-SC and none in CSH-IM, suggesting partial loss of differentiation or an increase in serotonin catabolism during transplantation.     

Photodynamic therapy of human malignant melanoma xenografts in athymic nude mice

While photodynamic therapy (PDT) for cutaneous malignancies including dermal recurrences of breast cancer and basal cell carcinomas has shown great promise, PDT of malignant melanoma has remained incompletely understood. A comparison study of the effects of PDT on human xenograft amelanotic and melanotic malignant melanoma in the athymic nude mouse model was performed. Twenty-four hours after ip administration of Photofrin II, the responses to total laser light doses of 25-300 J/cm2 were evaluated by histologic examination. Animals were also sacrificed 24 hours after administration of Photofrin II without light, and their uptake and localization of hematoporphyrin derivative (HpD) for each tumor were measured and compared. The results indicate that human xenograft melanotic melanoma, despite the fact that it contains more HpD than does amelanotic melanoma, is far less responsive to PDT. This result appears to be due to the competing chromophore melanin, which may inhibit the photochemical reaction at several key points.     

Control of human melanoma growth in nude mice by autologous allo-activated peripheral blood lymphocytes

Human peripheral blood lymphocytes (PBL) were activated in vitro by means of a pool of allogeneic PBL from normal donors and then evaluated for in vivo activity against human melanoma cells xenografted in splenectomized and irradiated athymic (nude) mice. The subcutaneous (s.c.) growth of human melanoma cells was inhibited by intravenous (i.v.) injection, 2 hr later, of such allo-activated, autologous and allogeneic PBL in 7/8 and in 6/9 mice respectively. Unstimulated PBL were ineffective. When allo-activated patients' lymphocytes were administered 3 days after s.c. implantation of autologous melanoma cells, inhibition of tumor growth was observed in 1/6 mice. A significant delay in tumor appearance was noted in the remaining animals. Unstimulated as well as allo-activated, lymphokine-releasing helper-enriched human PBL had no effect on melanoma xenografts, indicating that the tumor inhibition by tumor-cytotoxic allo-activated PBL was not due to recruitment of murine immuno-competent cells by human lymphokines. These results indicate that allo-stimulated, tumor-cytotoxic human PBL given i.v. to nude mice can circulate and inhibit the growth of autologous or allogeneic human melanoma cells implanted s.c.     

Inhibition of human tumor xenograft growth in nude mice by a novel monoclonal anti-HSPG isolated from human liver

Heparan sulfate proteoglycans (HSPGs) were isolated from normal human liver and α monoclonal antibody (MAb) was raised against them. Preliminary studies showed that MAb clone 1E4-1C2 was able to react with many cell lines tested, including hematopoietic cells and solid tumors. MAb1E4-1C2 was used to study whether HSPG was involved in growth and proliferation of human liver cancer using hepatocellular carcinoma (HCC) cell line (HepG2) as a model. Inhibition by MAb1E4-1C2 of HepG2 cell proliferation was studied in vitro by MTT assay. For in vivo assay, xenograft induction in athymic mice was performed. The results showed that MAb1E4-1C2 inhibited proliferation of HepG2 cells significantly, compared to isotype and medium control. MAb1E4-1C2 also suppressed the growth of tumor, resulting in smaller tumor size and weight. The investigation also showed that MAb1E4-1C2 inhibited proliferation and restricted tumor growth through the induction of apoptosis. The results suggest that HSPG might be involved in liver cancer cell proliferation. Therefore, a specific MAb that was raised against liver HSPG might be an alternative therapeutic agent for the treatment of human liver cancer.