Lethality of human melanoma cells for normal mice immunosuppressed with cyclosporin A

After intraperitoneal inoculation of BRO human melanoma cells, phenotypically immunocompetent mice given cyclosporin A (CyA) developed extensive and lethal tumors. Survivals were not significantly different for normal mice treated with this immunosuppressant compared to nude mice, which lack functional T-lymphocytes. BRO cells could be passaged in CyA-treated mice without alteration of isozymes or other properties tested. This model may have implications for growth and metastasis of human tumors under immunosuppressed conditions and may be useful for studying the mechanism of action of CyA in vivo.     

Complete inhibition of growth followed by death of human malignant melanoma cells in vitro and regression of human melanoma xenografts in immunodeficient mice induced by camptothecins.

The plant alkaloid camptothecin and three camptothecin derivatives were used to study responses of human malignant melanoma (BRO) cells xenografted in immunodeficient (nude) mice. Camptothecin and its derivatives 9-nitro-20(S)-camptothecin and 9-amino-20(S)-camptothecin inhibited growth of tumors and caused regression in all tumor-bearing mice. Tumor regression was accompanied by degenerative changes in the tumor cells, as assessed by microscopic observations of histological sections prepared from the tumors. No toxic effects were observed in the drug-treated mice, with or without xenografts. In parallel experiments, camptothecin, 9-nitro-20(S)-camptothecin, and 9-amino-20(S)-camptothecin inhibited proliferation of BRO cells in vitro and resulted in dramatic morphological cellular changes comparable to those observed in the sections of solid tumors. The derivative 12-nitro-20(S)-camptothecin had no effect on BRO tumors or cell cultures. The difference between 9-nitro-20(S)-camptothecin and 12-nitro-20(S)-camptothecin is the position at which the NO2 group is attached to the camptothecin molecule. In contrast to BRO melanoma cells, none of the camptothecin derivatives had any effect on cultured human melanocytes, the normal counterparts of melanoma cells. Taken together, the findings indicate that camptothecin and derivatives exert different effects on the growth and morphology of normal and malignant (BRO) melanocytes.     

Interactions with host macrophages and ability of human melanoma cell lines to grow in nude mice

The interactions of nude mouse macrophages with five human melanoma cell lines, characterized by their resistance to mouse NK activity and varying in their ability to grow s.c. in nude mice, were investigated. These lines were equally susceptible in vitro to both cytostatic and tumoricidal activities of activated peritoneal macrophages collected from nude mice inoculated 3 days previously with Brucella abortus B19R strains. I.p. injection of a poorly tumorigenic melanoma cell line (PTCL) in nude mice was followed by the local appearance of macrophages able to kill these cells in a 48-hr 3H-thymidine cytotoxicity assay. The level of tumoricidal macrophages was maximum for the first week and then slowly declined to disappear by the 4th week following PTCL inoculation. The use of an HTCL instead of a PTCL also induced macrophages able to kill HTCL cells, but the cytotoxicity level was lower and the activity disappeared more rapidly. In cross-experiments using PTCL-activated macrophages as effectors on HTCL targets, these cells were found to be less sensitive than PTCL cells when macrophages were taken at weeks 2 and 3 following PTCL inoculation. To investigate whether tumoricidal macrophages activated in vivo with human melanoma cells could also act in vivo, we inoculated these s.c. into nude mice, simultaneously with live HTCL cells. Peritoneal cells rich in melanoma-activated macrophages prevented HTCL growth in most recipients, whereas spleen cells from the same donor mice did not modify the tumor take. These data indicate that xenogeneic tumors could activate nude mouse macrophages in vivo and suggest that the ability of human tumors to grow in nude mice could be related to their capacity to activate host macrophages locally and to the susceptibility of human tumor cells to the tumoricidal activity of activated macrophages.     

Enhanced tumorigenicity, melanogenesis, and metastases of a human malignant melanoma after subdermal implantation in nude mice

Transplantation of human tumors into the organ or tissue of their origin (orthotopic transplantation) in nude mice can result in significant enhancement of tumor growth and metastases, compared with sc (ectopic) transplantation. Because melanocytes are normally found in the epidermal-dermal junction, intradermal inoculation of melanoma cells might be expected to improve their potential for malignant growth as xenografts. The purpose of our study was to examine this possibility. We found that because mouse epidermis and dermis are so thin, it was not possible to inject a bolus of tumor cells intradermally; instead the cells were actually deposited in the most superficial layer of the subcutis (i.e., subdermally). We evaluated the behavior of cells from a human melanoma cell line after sc or subdermal inoculation into National Institutes of Health Swiss athymic nude mice. The cells used were from (1) the predominantly amelanotic human malignant melanoma cell line MeWo, originally established from a melanotic lymph node metastasis, and (2) two MeWo variants resistant to wheat germ agglutinin (WGAr), which were selected for altered malignant capacities. Whereas 5 X 10(5) MeWo cells were required to achieve 100% tumor take with sc injection, only 2 x 10(4) cells were required with subdermal inoculation. Subdermal injection of the MeWo cells resulted in the development of highly melanotic and nonencapsulated primary tumors, which grew quickly into the dermis and epidermis and metastasized at high frequency to draining lymph nodes. In contrast, the tumors that developed after sc injection were found in the deepest layer of the subcutis and were predominantly amelanotic and encapsulated; they rarely metastasized to lymph nodes.(ABSTRACT TRUNCATED AT 250 WORDS)     

Specific organ metastases of human melanoma cells injected into the arterial circulation of nude mice

The purpose of this study was to examine whether organ specific metastasis can be produced by different subpopulations of cells isolated from a single human melanoma. Three clones, SB1, SB2, SB3, and a variant line SB1 Asc (obtained from ascites fluid after i.v. inoculation of SB1 cells in a nude mouse) - all isolated from a primary human cutaneous melanoma - were inoculated into the left ventricle (i.c.) or intravenously (i.v.) into nude mice. Three to 16 weeks later, the animals were killed and autopsied. The clones produced different patterns of metastasis. Three to 4 weeks after i.c. inoculation of 2 x 10(6) cells, cells of clone SB1 produced numerous rapidly progressing metastases in the brain, whereas cells of SB1 Asc produced large metastases in the adrenals and sometimes in the ovaries. Cells of clones SB2 and SB3 produced a few small metastases without any characteristics pattern. The initial arrest of tumor cells in various organs did not predict the outcome of metastasis. This conclusion is based on studies of distribution and fate of tumor cells labeled with 111Indium Oxine. Therefore, organ specific metastasis produced by human melanoma cells inoculated i.c. into nude mice is likely to depend on the interaction of unique metastatic cells with organ environment.     

Establishment and characterization of a human melanoma cell line (MV3) which is highly metastatic in nude mice

To select human melanoma cells that are highly tumorigenic and metastatic in nude mice we have implanted fragments of a fresh human melanoma metastasis subcutaneously (s.c.) into a nude mouse. After 3 passages in nude mice, part of the xenograft was cultured and a new melanoma cell line, MV3, was established. After intravenous (i.v.) inoculation of 2 x 10(6) MV3 cells, 95% of the nude mice (n = 20) developed lung colonies within 6 weeks. S.c. inoculation of 2 x 10(6) MV3 cells resulted in 95% tumor take, while 90% of the mice (n = 20) showed spontaneous metastases in the lungs within 7 weeks. Histological and immunohistological features of the original tumor of the patient were largely retained in the tumors of the mice and in the cell line in vitro. As shown by Alcian blue staining, MV3 cells contain large quantities of glycosaminoglycans (GAGs) and/or proteoglycanes (PGs), both in vivo and in vitro. The cells showed a marked expression of transferrin receptor, ICAM-1, EGF-receptor, and VLA-2 integrin. As only few human melanoma cell lines are available that frequently show metastasis in nude mice, the highly metastatic MV3 cell line represents a useful tool for studying the expression and regulation of molecules on human melanoma cells involved in the process of metastasis.     

Meningeal gliomatosis model in nude mice

The nude mouse has been proposed as a model animal for testing the chemosensitivity of human cancer cells, and encouraging results have been obtained. Murine brain tumor models were successfully prepared by direct inoculation of glioma cells into the cerebral hemisphere, an immunologically privileged area. However, the blood-brain barrier was destroyed mechanically by this manipulation. Accordingly, experimental models of MG were established by intracisternal inoculation of human glioma cells (ONS-6, -12, and -16). These cell lines were established by primary explant technique from our biopsy specimens. The outgrown cells were trypsinized and cultured in monolayers. Tumor cells (10(7) in 0.1 ml) were inoculated transcutaneously into the cisterna magna of BALB/c nu/nu mice using a 27-gauge needle. Tumor growth generally occur in no more than 10-20% of nude mice transplanted subcutaneously. Also, nude mice which were transplanted with these 10(7) glioma cells rejected the tumor. MG models which were inoculated with the same tumors intrathecally, however, all died within a month after inoculation. In in vitro studies, the doubling time (DT) of ONS-6, -12, and -16 were 30.5 h, 41.5 h and 60.9 h, respectively. Median survival time (MST) of MG models inoculated with ONS-6, -12, and -16 glioma cells were 8.5 days, 15.5 days and 18.5 days, respectively. These DTs were well correlated with the MST. Clinicopathological features observed in MG models were similar to those seen in diffuse leptomeningeal involvement of glioma in human beings. Tumor cells were disseminated in the diffuse arachnoid space, involving the cerebral hemisphere, spinal cord and throughout the cauda equina. Some tumor cells infiltrated the cerebral parenchyma and Virchow-Robin space. The models will be useful for investigating the pathophysiology of MG and the efficacy of chemotherapeutic agents.     

Growth of human melanoma in nude mice: suppression by T-lymphocytes from the tumor donor: brief communication.

Human malignant melanoma cell lines established in tissue culture were successfully transplanted sc into BALB/c nude mice. The growth rate of the resulting tumors was significantly suppressed when lymphocytes from the patient in whom the tumor arose were injected iv into BALB/c nude mice at the same time as sc tumor transplantation, but inoculation with lymphocytes from a person without a tumor was ineffective. Cell separation identified T-lymphocytes as the active subpopulation. Growth of the tumors was also significantly suppressed by reconstitution of the mice with normal BALB/c lymphocytes; lymphocytes from BALB/c mice previously immunized against the melanoma line were not more effective than nonimmune lymphocytes in preventing tumor growth. Sera from normal BALB/c mice or BALB/c mice immunized against a human melanoma cell line effectively suppressed growth of that cell line in BALB/c nude mice if given at the time of tumor transplant. The results suggested that, whereas murine lymphocytes reconstitute the ability of nude mice to react to xenogeneic antigens on the human tumor, human lymphocytes showed greater specificity to autologous human melanoma-associated antigens.     

Extrapulmonary, tissue-specific metastasis formation in nude mice injected with FEMX-I human melanoma cells

FEMX-I human malignant melanoma cells, originating from a lymph node metastasis in a patient, uniquely and selectively produced extrapulmonary metastases after i.v. injection of cells prepared from xenografts into adult, nude mice. After a lag time of approximately 50 days, metastases were observed in s.c. sites at the back and front of the neck, and in axilla and inguinal regions. Tumor colony formation in lungs were never detected. The interscapular tumors showed a close relationship to brown fat, partly infiltrating this tissue, whereas the other s.c. tumors seemed to be localized to lymph nodes. Mesenterial and mediastinal lymph node metastases were frequently found, together with retroperitoneal tumors along the spine. The normal cells of the adrenal medulla were often replaced by melanoma cells, whereas the cortical tissue was not affected. The conclusion that FEMX-I cells possess an inherent ability for tissue-specific metastasis formation is supported by the metastatic pattern seen after i.p. and intrasplenic injection, as well as after inoculation of the cells in the footpads of the mice. The relatively slowly growing FEMX-I tumors showed the same differentiated morphology as the patient's tumor, independent of the site of growth and the number of passages in the animals. The FEMX-I tumor was easily established as a cell line in vitro. Such cells showed a strongly reduced metastatic capacity, indicating that the in vitro growth conditions had induced alterations in the FEMX-I cells influencing their ability to form site-specific metastases, changes that were shown to be reversible. It is suggested that structures on the surface of the tumor cells, as well as growth factors in the host tissues, may be of importance for the observed tissue specificity. The FEMX-I melanoma, which, as a human tumor in nude mice, has a unique metastatic pattern, offers possibilities for investigating mechanisms involved in site-specific metastasis formation, as well as for testing effects of antimetastatic, chemotherapeutic, and immunotherapeutic agents against human extrapulmonary micro- and macrometastases.     

ESTABLISHMENT OF A HUMAN PANCREATIC ADENOCARCINOMA CELL LINE （JF305） WITH p53 EXPRESSION

李昕，王宏，姜奕，王晓华，贾兰玲，张宝庚ＥＳＴＡＢＬＩＳＨＭＥＮＴＯＦＡＨＵＭＡＮＰＡＮＣＲＥＡＴＩＣＡＤＥＮＯＣＡＲＣＩＮＯＭＡＣＥＬＬＬＩＮＥ（ＪＦ３０５）ＷＩＴＨｐ５３ＥＸＰＲＥＳＳＩＯＮ￥ＬｉＸｉｎ；ＷａｎｇＨｏｎｇ；ＪｉａｎｇＹｉ；Ｗａｎｇ...     

Establishment and characterization of a carcinoembryonic antigen (CEA)-producing cell line from a human carcinoma of the exocrine pancreas

A human carcinoembryonic antigen (CEA)-producing cell line, T3M-4, has been established from explant cultures of a primary human pancreatic exocrine adenocarcinoma transplanted into nude mice. The tumor had metastasized in the patient. The tumor obtained from metastatic lymph nodes was the initial source for implantation in athymic nude mice. In the primary culture, host fibroblasts were eliminated by the use of the antiserum raised against nude mouse cells. T3M-4 cells have been continuously propagated in vitro during the past 26 months. The cells grew in a monolayered sheet with about 31 hours of population doubling time. The cells exhibited epithelial morphologic features resembling the structure of the original tumor, and they showed tumor takes when inoculated into athymic nude mice. Xenografts established from the cell line have retained a similar histology to the original tumor on serial transplantation. Chromosomal analysis revealed the cell line to be a human aneuploid one with a hyperdiploid mode. T3M-4 cells possess the characteristic function of CEA secretion in vitro in culture and in vivo in nude mice bearing the tumors produced by inoculation with the cultured cells. In view of these characteristics, T3M-4 cell line represents a new human pancreatic exocrine adenocarcinoma cell line that produces CEA.     

Effectiveness of anticancer drugs determined in nude mice inoculated with [125I]5-iodo-2'-deoxyuridine-prelabeled human melanoma cells

Anticancer drugs were tested on NIH-2 nude mice inoculated ip with BRO human melanoma cells, which are rapidly lethal for these hosts. Criteria for drug activity were a) increased host survival and b) an increased rate of radioactivity loss from mice bearing BRO cells prelabeled with [125I]5-iodo-2'-deoxyuridine. Diphtheria toxin, which is selectively toxic to human cells compared to mouse cells, prolonged host survival and accelerated 125I elimination in a dose-dependent manner. Drugs that increased the rate of 125I loss compared to the rate of untreated mice also prolonged the lives of treated mice. With one exception, drugs that did not accelerate 125I elimination had little or no effect on the length of survival.     

Establishment and characterization of a pancreatic carcinoma cell line derived from malignant pleural effusion

BACKGROUND/AIMS:A novel cell line, designated p34, was developed from the malignant pleural effusion of a patient with carcinoma of pancreas. The objective of this work was to characterize this cell line. METHOD: The in vitro studies included karyotype analysis, immunohistochemistry, XTT cell proliferation assay, analysis of the cell cycle by FACS and cell sensitivity to chemotherapeutic drugs and irradiation. Subcutaneous and intra-spleen inoculations into nude mice were carried out to study the tumorigenicity and the metastatic tendency of this cell line. RESULTS: The p34 cell line showed typical morphological characteristics of epithelial pancreatic tumor cells. The cells were hyperdiploid with a modal number of 48, and had two markers, deletion in the short arm of chromosome 2 and duplication of the short arm of chromosome 8. The doubling time was 16 h. Subcutaneous inoculation of the cells into nude mice yielded 100% tumorigenicity, and intra-spleen inoculation resulted in extensive intra-abdominal spread. The antiproliferative effect of chemotherapy (gemcitabine, cisplatin, taxol and vinorelbine), chemopreventive agents (celecoxib and curcumin) and radiotherapy showed dose-dependent cytotoxicity. CONCLUSIONS: This p34 cell line can be used as a new model for studying various aspects of the biology of human pancreatic cancer and potential treatment approaches for the disease.     

人原发性小肠恶性黑色素瘤裸鼠原位移植高转移模型的建立

目的 建立人原发性小肠恶性黑色素瘤裸小鼠原位移植高转移模型.方法 将手术切除的人原发性小肠恶性黑色素瘤原发灶和肝转移灶新鲜瘤组织块分别植入裸鼠小肠黏膜层内,观察原位移植的成瘤率、移植瘤的侵袭性和转移率,并进行形态学、流式细胞分析和染色体核型分析.结果 人小肠恶性黑色素瘤原发灶和肝转移灶新鲜组织均移植成功,建成人原发性小肠(原发灶)恶性黑色素瘤裸鼠原位移植高转移模型(ttSIM-0602)和人原发性小肠(肝转移灶)恶性黑色素瘤裸鼠原位移植肝转移模型(HSIM-0603).HSIM4)602和HSIM-0603模型分别传至21代和23代,共移植裸鼠227只,其肿瘤移植生长率和液氮冻存复苏成活率均为100%.HSIM-0602模型肝转移率为65.7%,肺转移率为84.8%,淋巴结转移率为63.8%.HSIM-0603模型肝转移率为100%,肺转移率为46.7%,淋巴结转移率为71.3%.移植瘤组织病理学为小肠高度恶性黑色素瘤.免疫组织化学显示,S-100蛋白和HMB-45均为阳性表达.电镜下,瘤细胞浆内可见大量的黑色素小体,也可见黑色素复合体.HSIM-0602模型移植瘤细胞DNA指数为1.59±0.07,HSIM-0603模型移植瘤细胞DNA指数为1.71±0.12,均为异倍体.染色体核型分析显示,HSIM-0602模型移植瘤细胞染色体数为55～57条,HSIM-0603模型移植瘤细胞染色体数为57～59条.结论 HSIM-0602和HSIM-0603模型是成功的人原发性小肠恶性黑色素瘤裸鼠原位移植自发性高转移模型,完整地模拟了人原发性小肠恶性黑色素瘤患者的自然临床病理过程,为研究原发性小肠恶性黑色素瘤转移生物学和抗转移治疗提供了理想的动物模型.     

Highly pigmented human melanoma variant which metastasizes widely in nude mice, including to skin and brain

The properties of a highly malignant human melanoma variant cell line which metastasizes in nude mice in a tissue-specific pattern are described. The variant, called 70-W, was isolated from the MeWo malignant melanoma by exposure of the latter to stepwise increasing concentrations of the toxic lectin, wheat germ agglutinin. After nine cycles of treatment a population of wheat germ agglutinin-resistant cells was obtained that manifested a 4-fold resistance to wheat germ agglutinin, a property which was found to be stable in culture for over 6 months in the absence of the lectin. Intravenous inoculation of 70-W cells into 4-6-week-old nude mice revealed remarkable differences in metastatic (organ colonization) behavior. Whereas the parent MeWo cells gave rise only to lung metastases, most of which were amelanotic, injection of the 70-W cells resulted in multiple skin (s.c.) and brain and, to a lesser extent, bone marrow, ovarian, mesenteric (gut-associated), muscle, and abdominal metastases all of which were highly melanotic. This is the first report of brain metastases of a human tumor in nude mice. They were found to be bilateral and confined to the deeper layers of the cerebral cortex. The unique malignant behavior of 70-W cells in nude mice should facilitate studies of host and tumor cell factors involved in human melanoma metastasis, melanogenesis, and development of new treatment strategies for disseminated human malignant melanoma.     

Establishment of human glioma cell line--nude mice solid tumor model NHG-1 and its characteristics

Establishment and its characteristics of a nude mice solid tumor model NHG-1 from human glioma cell line are reported. 5-8 week old NC nude mice of both sexes and SHG-44 cell line used in this experiment were from our laboratory. The initial successful transplantation rate was 7/11 (64%) and that of 30 passages in the subsequent 4 years was 100%. After subcutaneous inoculation, growth curve showed a latent period in week 1-2, slow growing period in week 3-4, rapid growing period in week 5-6 and a final plateau period in week 7. The doubling time was 7 days and cell cycle time was 2.5 days. The cells in G1, S and G2M phases comprised 56%, 27% and 17%, respectively. The survival time of the host was 54 +/- 15 days. The tumor tissues showed a tendency towards invading the surrounding soft tissues. By morphological observation with light and electron microscopes, LDH isozyme assay, PAP immuno-histochemistry labelling GFAP and chromosome analysis, it is confirmed that the transplantable tumor possesses the characteristics of human malignant glioma. The estrogen receptor in the transplantable tumors demonstrated by cytochemical assay indicates that the glioma carcinogenesis is related to endocrine factor of the host. The therapeutic effects of anticancer drugs, such as ACNU, BCNU and 10-hydroxy-2-decenoic acid from the royal jelly on NHG-1 model are evaluated.     

Regression of Experimental Human Leukemias and Solid Tumors Induced by Epstein-Barr Virus-Immortalized B Cells

We previously have reported on an experimental athymic mouse model in which regression of human Burkitt's lymphoma is induced by either coinjection with or intratumor inoculation of Epstein-Barr virus (EBV)-immortalized human B cells. In the current study, we were interested in determining whether the powerful antitumor effects of EBV-immortalized B cells could be effective against a variety of human tumors grown in athymic mice, including acute lymphocytic leukemia, malignant melanoma, acute promyelocytic leukemia, neuroblastoma, lung carcinoma, colon adcnocarcinoma, Wilms tumor, Hodgkin's lymphoma, rhabdomyosarcoma and breast adenocarcinoma. We report here the results of experiments in nude mice that demonstrated the potent antitumor effect of EBV-immortalized B cells against human tumors derived from a variety of different tissues.     

Characterization of a metastasis-deficient lectin-resistant human melanoma mutant

A mutant (3S5), almost completely deficient in pulmonary metastatic ability in nude mice, was recently isolated from a human malignant melanoma cell line (MeWo). The WGA-resistance (WGAr) phenotype of 3S5 cells was accompanied by a collateral hypersensitivity to the lectin BSII (Bandeiraea simplicifolia), similar to that observed previously in class-I non-metastatic WGAr recessive genetic mutants isolated from a highly metastatic mouse tumor. The lectin-resistance/sensitivity profile of 3S5 cells was completely stable in tissue culture for 9 months when the cells were grown in the absence of WGA. Twenty independent clones of 3S5 were analyzed and all manifested a similar lectin-resistance pattern, and severe deficiency in pulmonary metastatic ability following i.v. or s.c. inoculation, when compared to the parent MeWo cells. Some heterogeneity in metastasis was noted, especially with respect to occasional extrapulmonary spread. The majority of the small number of metastases obtained after i.v. inoculation of 3S5 cells maintained the lectin-resistance profile and deficient metastatic potential characteristic of 3S5 cells upon re-inoculation into other nude mice. The results provide further evidence that cell-surface glycosylation changes can significantly after metastatic potential, including that of human tumors.     

Reconstituted basement membrane (matrigel) enhances the growth of human glioma cell lines in nude mice

Transplantation of human cancers into immunologically deficient mice is widely used to study potential therapeutic interventions in vivo. For brain tumor research, however, several factors limit more widespread application of this animal model. First, only a minority of human glioma-derived cell lines are tumorigenic in nude mice. In addition, even for tumorigenic cell lines, tumor take is variable and growth is often slow for tumors derived from cell inoculums. Reconstituted components of tumor basement membrane (matrigel) have been found to improve the growth in nude mice of several types of human tumors originating outside the central nervous system when premixed with the tumor cells before subcutaneous inoculation. We investigated the ability of matrigel to enhance the growth in nude mice of tumors derived from the human glioma cell lines U-251 MG, U-373 MG, SNB-78 and SNB-101.Athymic nude mice (NIH Swiss background, nu/nu genotype) were inoculated subcutaneously with 1.0 × 10⁶ tumor cells alone or after premixing with an equal volume of liquid matrigel. U-251 and U-373 cells were tumorigenic, with palpable tumors present by about 2 to 3 weeks. Co-injection of these cell lines with matrigel resulted in higher tumor-take rates, from 6/10 to 8/8 animals for U-251 at 60 days, and from 9/12 to 11/11 animals for U-373 at 60 days. Matrigel also enhanced tumor growth, with tumors at 45 days significantly larger than those formed in the absence of matrigel, for both cell lines (p < 0.01). SNB-78 and SNB-101 cells did not give rise to progressively enlarging solid tumors with or without matrigel.Matrigel enhances the growth of tumorigenic human gliomas in athymic nude mice. This technique provides a model with more consistent tumor take and more rapid growth kinetics for human glioma cell lines that are tumorigenic in nude mice.The U.S. Government right to retain a non-exclusive, royalty free licence in and to any copyright is acknowledged     

Albendazole: a potent inhibitor of vascular endothelial growth factor and malignant ascites formation in OVCAR-3 tumor-bearing nude mice.

PURPOSE: Angiogenesis and vessel hyperpermeability are the two factors leading to the formation of ascites. Vascular endothelial growth factor (VEGF) plays a pivotal role in malignant ascites formation. We have recently shown that albendazole inhibits peritoneal growth of human colorectal cancer cells (HT-29). The present study was designed to find out if albendazole can suppress ascites formation in ascites-producing peritoneal carcinomatosis. EXPERIMENTAL DESIGN: Female nude mice bearing peritoneal tumors of human ovarian cancer cells (OVCAR-3) were treated with albendazole. Following i.p. inoculation and ascites development, mice were given i.p. albendazole (150 mg/kg) or the vehicle x 3 weekly for 4 weeks. RESULTS: Whereas vehicle-treated mice developed overt ascites requiring repeated aspiration, ascites formation in the albendazole-treated mice was markedly suppressed. As a result of this, 7 of 10 mice from the control group had to be euthanized before the course of treatment was over. Suppressed ascites production and reduced tumor vascularity observed was a result of dramatic reduction in tumor VEGF production as revealed by profoundly lower VEGF ascites fluid and plasma levels. In vitro, incubation of SKOV-3 cells with various concentrations of albendazole led to significant dose-dependent inhibition of VEGF secretion. Examination of floating tumor cells collected from the peritoneal wash revealed profound down-regulation of VEGF mRNA in albendazole-treated mice. CONCLUSIONS: These findings suggest for the first time that in nude mice bearing OVCAR-3 peritoneal tumors, by inhibiting VEGF production, albendazole abolishes tumor angiogenesis and ascites formation.