Rapid Growth and Spontaneous Metastasis of Human Germinal Tumors Ectopically Transplanted into Scid (Severe Combined Immunodeficiency) and Scid-nudestreaker Mice

Xenograft acceptance, growth and spontaneous metastasis of ectopically transplanted human germinal tumors were compared among scid mice, athymic nude mice and F₂ hybrids constructed from scid and nude mice, in relation to the impairments of T and B cell functions in these mice. In scid mice which are deficient in T and B cell functions, human yolk sac tumor (YST-2) that originated from the ovary grew to enormous sizes in 100% of the animals after both subcutaneous and intraperitoneal transplantation, while only half (59.1% and 51.9%) of the subcutaneous and none of the intraperitoneal transplants were accepted in usual athymic nude mice (BALB/c- nu/nu and CDl- nu/nu ). The YST-2 grew rapidly in scid mice, developing 3 to 10 times larger tumors compared to nude-streaker (AKR/J- nu^str/nu^str ) and usual nude mice, respectively. Furthermore, ectopically transplanted tumors spontaneously metastasized to distant organs (mostly to the lung) in scid mice (but less frequently in leaky scid mice), while metastases have never been found in nude mice. Although a xenograft of human classic (typical or pure) seminoma of the testis has never been established in nude mice, it grows slowly in one-third (36.4%) of scid mice and very rapidly in all of scid-nu ( scid/scid; nu^str/nu^str ) double mutant mice. Spontaneous metastases of xenografted seminomas were also observed in distant organs (lymph node, lung, liver, spleen, and kidney). The metastastic distribution of the two human germinal tumors in scid and scid- nu^str mice mimics that found in human. These results (xenograft acceptance, growth of transplanted tumors and degree of metastatic spread) were compatible with the level of T and B cell impairments indicated by FACS analysis, as well as mitogen responses, serum IgG and morphological features of the thymus.     

Giant cell carcinomas of the lung producing colony-stimulating factor in vitro and in vivo

Two culture cell lines (C-Lu65, C-Lu99) were established from human giant cell carcinomas of the lung transplanted in athymic nude mice (BALB/c, nu/nu). During early passage in tissue culture, C-Lu65 grew as a loosely adherent monolayer with some piling-up and with floating cells. After 30 successive subcultures, C-Lu65 began to grow in suspended cell clusters, showing a faster growth rate. C-Lu65 was characterized by multinucleated giant cells with large abnormal nuclei and prominent nucleoli. C-Lu99 grew as adherent cells, and fewer multinucleated giant cells were observed. C-Lu65 and C-Lu99 showed some ultrastructural differences in cell surface and cytoplasmic features. Chromosomal analysis revealed numerical and structural abnormalities in both cell lines. Cell-free supernatants from both cell lines stimulated the colony formation of mouse bone marrow cells in vitro. In addition, mice bearing tumors induced by transplanting C-Lu65 and C-Lu99 showed remarkable leukocytosis without evidence of infection. These results suggest that these two cell lines release colony-stimulating factor both in vitro and in vivo.     

Development of androgen-independent spindle cell tumors from androgen-dependent medullary Shionogi carcinoma 115 in androgen-depleted nude mice.

When Shionogi carcinoma 115 (SC115, undifferentiated medullary carcinoma showing compact cell pattern and containing androgen receptor) was transplanted into male and female DS mice, it grew only in males. In contrast to this strict androgen dependency in DS hosts, tumors composed of spindle-shaped cells appeared in more than 80% of cases when SC115 tumor was inoculated into female or castrated male nude athymic (BALB/c-nu/nu) recipients. These spindle cell tumors neither contained cytosol androgen receptor nor showed biologically defined androgen dependency. As spindle cell tumors could be serially transplanted in DS mice but not in BALB/c-+/+ mice and as the original SC115 (medullary carcinoma showing a compact cell pattern) tumor and the spindle cell tumor had many identical chromosome abnormalities, these two types of tumors seem to have a common origin in spite of their morphological, biochemical, and biological differences. Since spindle cells could not be detected histologically in SC115 tumors maintained in intact male DS mice, the present results seem to suggest that SC115 cells may change their morphological, biochemical, and biological characteristics within one passage in androgen-depleted nude athymic mice.     

Influence of the mouse hepatitis virus (MHV) infection on the growth of human tumors in the athymic mouse.

The growth characteristics and histological appearance of tumors resulting from transplantation of the tumor lines HEp-2 and SW480 into pathogen-free and mouse hepatitis virus infected athymic mice were studied. Subcutaneous or intraperitoneal implantation 1 x 10(6) neoplastic cells into pathogen-free animals resulted in tumor growth. Subcutaneous transplants grew locally, surrounded by a capsule of connective tissue. The fibrovascular stroma supporting the neoplastic tissue was minimal and infiltration of tumor capsule was observed. Intraperitoneal tumors grew in a multifocal pattern, were not encapsulated, showed marked invasiveness and metastasized. The same number of neoplastic cells (1 x 10(6)) transplanted into hepatitis-positive animals failed to develop into grossly visible tumors. When the number of transplanted cells was increased to 2 x 10(7), tumors appeared in a few animals. All tumors, regardless of the site of transplantation, were characterized by the presence of severe fibrohistiocytic reaction at the site of implantation that possibily influenced the tumor growth. No evidence supporting T-cell-mediated tumor rejection was observed. It is concluded that the state of health of the athymic mice is critical for the growth of human tumors and may account for the variations in reporting successful transplantation of such tumors in nude mice.     

四株人体和大鼠食管癌裸鼠移植瘤株的建立及其生物学特性研究

我们将12例人食管癌手术切除组织接种于NC和BALB/C裸鼠皮下,其中2例获移植成功,裸鼠间传代达32代,历时2年余,建立了二株人食管癌裸鼠移植瘤,分别为人食管鳞状细胞癌裸鼠移植瘤(HEC_2)和人食管腺鳞癌裸鼠移植瘤(HEC_6)。两移植瘤具有潜伏期短、移植成功率高(100％)、荷瘤存活时间长及生物学特性稳定等特点。并在建立大鼠食管癌细胞系(RE25—3.)的基础上,建立了二株大鼠食管癌裸鼠移植瘤(RE25/N和RE66/N)。此四株人体和大鼠食管癌裸鼠移植瘤株为食管癌病因、发病机理及治疗的研究,提供了极好的模型。     

Progressive growth of a human pleural mesothelioma xenografted to athymic rats and mice

A human malignant pleural mesothelioma was xenografted serially in athymic nude Rowett rats for 27 passages during 33 months. After the two initial passages (P), the take rate during P3-9 was 100% (192/192). The tumour grew progressively during P3-9 in 99% (190/192) and regressed totally in 1% (2/192). The take rate for the tumour xenografted to athymic BALB/c mice was also 100% (17/17) and no regressions were observed. During serial passaging in nude rats, the tumour-volume doubling time (TD) decreased from 6 days in P2 to 3 days in P8-9 (P less than 0.001) and then remained around 3 days during P10-25. A TD of 11 days in P1 (man-mouse) for tumours grown in mice decreased during 10 passages in rats to 4 days (P less than 0.005) when the tumour was transplanted to mice in P11. Light microscopic morphology of the tumour was retained in rats and mice. We believe that our experimental tumour model using the nude rat as a carrier of the xenograft will be useful for studies of human mesothelioma.     

Human brain tumor transplantation into nude mice.

Seven human brain tumors were transplanted into the brains (6/7 takes) and subcutaneous tissues (7/7 takes) of athymic nude mice. Compared to experimental animal brain tumors, these tumors, taken directly from patients in the operating room and transplanted, grew more slowly in the mice; their growth rates following explant generally paralleled those in the patients. A rough correlation was seen between the degree of the tumor's malignancy and both successful take and rate of growth following explant. The tumors' growth rates increased during serial transplantation after explant. Two tumors developed into long-term serial lines; both came from gliosarcomas. Preliminary chemotherapy experiments with these two lines demonstrated different chemosensitivities. One line was very sensitive to the nitrosoureas and resistant to procarbazine; the other line was more sensitive to procarbazine than to the nitrosoureas. This model permits study of the biologic behavior of human brain tumors growing intracerebrally and subcutaneously in nude mice.     

裸鼠的人原发性胃癌移植瘤生物学及分子遗传学性质研究

A human primary gastric cancer tissue (adenocarcinoma II-III) was transplanted into nude mice (SWISS/DF. nu/nu). It has been transferred for 8 generations at 56 sites in 28 nude mice with transplantable rate of 100%. The transplanted tumor is designated as transplantable human primary gastric cancer-1 in nude mice (THPGC-1). The growth of THPGC-1 is rather rapid and the size of transplanted tumor reaches 1 cm2, 4-5 weeks after transfer. The morphology and histochemistry of the original tumor were retained well in the initial and serial transplanted tumors. THPGC-1 could secret carcinoembryonic antigen (CEA). After intravenous or intraperitoneal injection of 131I-antiCEA monoclonal antibody into the THPGC-1 bearing nude mice, the radiolabeled antibody was concentrated and localized in the tumor as shown by gamma-camera analysis. Similar pattern of lactate dehydrogenase isoenzyme was observed both in primary gastric cancer tissue and THPGC-1 tissue. Chromosomal examination revealed that THPGC-1 was human aneuploid ones. Southern blot analysis showed that the pattern of repetitive DNA bands and the structures of 28s, rDNA, c-H-ras and c-myc genes in THPGC-1 were identical to the original primary gastric cancer DNA. The results suggest that THPGC-1 be a reliable model for the research of the molecular biology of cancer cells and experimental gastric cancer diagnosis and treatment.     

荷人前列腺癌裸鼠移植瘤模型的建立及其应用研究

目的 应用人前列腺癌细胞株LNCaP接种裸鼠,鼠间移植传代,建立人前列腺癌裸鼠移植瘤模型.方法 观察移植瘤大体形态和组织病理学检查,并应用免疫组织化学方法观察肿瘤组织细胞中前列腺特异膜抗原（PSMA）的表达以及125I标记抗PSMA单抗的荷瘤裸鼠体内放射免疫显像.结果 移植瘤的形态和功能特性与原发肿瘤基本相似,移植瘤的移植成功率为100%;放免显像显示标记抗体能浓聚于肿瘤部位,经尾静脉注射标记抗体后96 h,肿瘤显像清晰,肿瘤组织/非肿瘤组织放射性比值（T/NT）均大于2.8.结论 本动物模型的建立为今后前列腺癌放射免疫治疗的实验研究提供1个理想的模型.     

Residual immunity of athymic NCr/Sed nude mice and the xenotransplantation of human tumors

This study assessed the residual immunity possessed by NCr/Sed (nu/nu) athymic nude mice and examined strategies to reduce it in order to enhance the transplantability of human tumors for experimentation. Adult (8-week-old) female mice had fewer T cells (11%) and more B and NK (asialo-GM1-positive [ASGM1+]) cells in their spleen than euthymic (nu/+) controls. The number of phenotypically mature T cells increased with age, peaking at 16 weeks. ASGM1+ cells also increased in number over time, although the NK-activity decreased after 12 weeks. B cells remained relatively constant in number. Athymic NCr/Sed nude mice displayed reactivity against a human squamous carcinoma xenograph (FaDu), in a Winn's test and TD50 assay. Immunity against xenografts (TD50 assay) was significantly lower (by a factor of 2) in 4-week-old than in 12-week-old nude mice. Similarly, a significant 2-fold reduction in TD50 was obtained after a single intraperitoneal injection of cyclophosphamide into 8-week-old animals. Chronic (greater than 8 weeks) exposure of the nude mice to subcutaneously administered beta-estradiol markedly reduced the number of splenic NK cells and their cytolytic activity, but the TD50 reduction was not statistically significant (p = 0.1). Six Gray whole-body irradiations (WBI) had been shown to produce a highly significant, 3-fold reduction in the TD50 for FaDu. Flow cytometric analysis of splenic lymphoid cells from whole-body-irradiated recipients revealed: (a) marked initial depletion in the absolute numbers of lymphoid cells; (b) marked and long-lasting depletion of T cells, with slow and minimal recovery only evident between 6 and 12 weeks; (c) rapid, almost complete, depletion of B cells with prompt and partial recovery after 2 weeks; (d) depletion of NK cells and NK activity, with recovery by 10 weeks. No change in the number or phagocytic capacity of resident peritoneal macrophages was seen. These data give further support to a postulated role for residual T cells in the xenoreactivity of NCr/Sed nude mice.     

Chemotherapy of human yolk sac tumor heterotransplanted in nude mice

The chemotherapeutic effects of cis-diamminedichloroplatinum + vinblastine + bleomycin (PVB) on 3 human yolk sac tumors (YST-1, YST-2, and YST-3) of the ovary, which were heterotransplanted into BALB/c nude mice, were compared with the effects of vincristine + actinomycin D + cyclophosphamide (VAC), the combination currently favored for treatment of yolk sac tumors. PVB and VAC therapies were performed for 3 weeks. Both PVB and VAC significantly reduced the tumor volume of all the treated tumors. The mean weights of tumors in animals treated with PVB or VAC were, in percent of the mean tumor weight in untreated animals: 1.3 and 1.6 for YST-1, 2.5 and 3.3 for YST-2, and 5.5 and 2.7 for YST-3, respectively. A strong correlation was noted between tumor volume and alpha-fetoprotein level in the sera of mice bearing YST-1 or TST-2 tumors.     

Growth and metastasis of fresh human benign and malignant tumors in the head and neck regions transplanted into scid mice.

Surgically resected fresh human tumors (16 malignant and 1 benign) in the head and neck regions were s.c. transplanted into scid mice. All malignant tumors (12 squamous cell carcinomas, 2 papillary adenocarcinomas and 1 adenoid cystic carcinoma) except one heavily irradiated squamous cell carcinoma could grow in scid mice. However, all of three squamous cell carcinomas of the maxillary sinus grew only in 50% of scid mice, and two of them failed to grow in the second transfer. The remainder were successfully transplantable for further generations, except one accidental case. A benign tumor, a follicular adenoma of the thyroid gland, was also accepted in scid mice and was transplantable for further generations, though its growth was very slow. Distant metastases were found in the lung only when poorly differentiated carcinomas were transplanted into scid mice, but did not occur until 90 days after the transplantation of well and moderately differentiated carcinomas. The histological characteristics of both malignant and benign tumors were retained well in all of the xenografts and metastatic lesions. Thus, scid mice seem useful to investigate not only the properties of benign and malignant human tumors but also the metastatic spread of tumors which threaten the life of cancer patients.     

Potential of the scid mouse as a host for human tumors

Summary Animal models of human tumors and their metastases that effectively mimic clinical disease are in considerable demand. While it is certainly true that athymic nude mice provide us with useful models to study a large number of human tumors in vivo, it is also well known that nude mice usually do not develop spontaneous metastases and are not suitable for all types of tumors. Therefore, the scid mouse that allows disseminated growths for a number of human tumors, particularly hematologic disorders and malignant melanoma, can be used preferentially for the investigation of such malignancies. The potential to study the interaction of human immune cells and human tumor in an in vivo model is another unique feature of scid mice that will add to their usefulness in experimental cancer research.     

Antitumor activity of murine tumor necrosis factor (TNF) against transplanted murine tumors and heterotransplanted human tumors in nude mice

The antitumor activity of highly purified tumor necrosis factor (TNF) was tested against eight kinds of murine tumor and five kinds of human tumor heterotransplanted into nude mice. Mice were treated by intravenous or intratumoral injection of TNF, commencing when the tumors were well established. TNF showed an excellent curative effect against all kinds of murine and human tumors tested. Meth A sarcoma, Colon 26, Ehrlich, sarcoma 180, MM 46, MH 134, B16 melanoma, and Lewis lung tumors transplanted into mice underwent tumor necrosis and regression following a single injection of TNF. Sometimes a complete cure was observed in Meth A sarcoma, sarcoma 180, Ehrlich, and MM 46 tumors. Human cancers, SEKI, HMV-I, KATO-III, MKN 45, or KB, heterotransplanted into nude mice, also exhibited tumor necrosis and regression in size following several intratumoral injections of TNF. A great difference in curative effects of TNF was observed in Meth A sarcomas between those transplanted into BALB/c nu/+ and into BALB/c nu/nu mice: following a single intravenous administration the effect was stronger in BALB/c nu/+ than in nu/nu mice. In contrast, tumor necrosis was almost the same in nu/+ and nu/nu mice following intratumoral administration. The present results thus indicate that TNF from mice had an antitumor activity against not only murine tumors but also human tumors. In addition to direct cytotoxicity against tumor cells, TNF induced a host-mediated factor which contributed to the antitumor effects.     

A comparative study on the xenotransplantability of human solid tumors into mice with different genetic immune deficiencies

These experiments set out to assess the role of NK and B cells in the resistance of nude mice to human tumor xenotransplantation. The transplantability of 9 fresh and 8 cultured human tumors was compared in 2 strains of mice with different genetic immune deficiencies: athymic NCr/Sed (nu/nu) nude mice, and nude-beige-xid (N:NIH-nu-bg-xid/Sed mice). Flow cytometric studies showed both strains to be deficient in Thy. 1.2 (T) cells and unresponsive to stimulation by Concanavalin A (Con A) or direct T-cell-receptor triggering with anti-CD3. The number of B cells was similar in the 2 strains, but the response to lipopolysaccharide (LPS) was markedly reduced in the nude-beige-xid animals. The number of asialoGM1-positive cells (predominantly NK) detected by flow cytometry was also reduced in the nude-beige-xid mice. The transplantability of the human tumors was found to be equivalent in the 2 strains. Quantitative cell-transplantation assays performed for 2 of the tumor cell lines did not reveal any subtle transplantation advantage for the more broadly immune-deficient animals. No evidence could, therefore, be found to suggest that NK or B cells were major determinants of human tumor xenotransplantability in these strains of mice.     

Tumorigenicity and dissemination of primary and metastatic human melanomas implanted into different sites in athymic nude mice

In the present study, the growth features and metastatic capacity of human primary and metastatic melanomas transplanted by different routes in nude mice was examined. Eight different human melanoma early cell cultures derived from 3 primary tumors, 1 local recurrence and 4 metastatic lesions of 6 melanoma patients were characterized for cell surface HLA (class I and II) and melanoma-associated antigens and karyotype. These tumors were then transplanted in CD-1 outbred nude mice by subcutaneous, intraperitoneal and intrasplenic routes. It was found that 7 out of 8 melanomas were tumorigenic after subcutaneous implantation without giving rise to metastases; 5 out of 7 melanomas grew when injected intraperitoneally and 3 of them disseminated to peritoneal organs and infiltrated intraperitoneal lymph nodes, liver and pancreas; of 8 melanomas implanted intrasplenically 4 grew in the spleen or invaded intraperitoneal lymph nodes, liver, pancreas, ovaries or lungs. Primary and metastatic melanomas did not differ in the pattern of dissemination. In fact, 2 out of 4 metastases and 1 of 3 primary tumors and 1 recurrence disseminated after intrasplenic or intraperitoneal inoculation. Heterogeneity in the growth pattern of different metastases of the same melanoma patient was also found. No correlation could be detected between the metastatic ability of melanoma cells studied and clinical stage of patients, tumor cell karyotype abnormalities, modal number or with the antigenic phenotype.     

二烯丙基二硫对人胃癌细胞裸鼠移植瘤的抗肿瘤作用

背景与目的:既往研究发现二烯丙基二硫(diallyldisulfide,DADS)在体外可抑制多种肿瘤细胞生长,但在体内抗肿瘤作用的研究报道较少。本实验旨在探讨DADS对人胃癌细胞移植瘤在BALB/C裸鼠体内生长的影响。方法:未经药物处理和经30mg/LDADS处理1天的胃癌细胞MGC803接种于裸鼠皮下;观察体外DADS处理MGC803细胞裸鼠移植瘤的成瘤情况和未处理MGC803细胞移植瘤成瘤后腹腔注射DADS对胃癌移植瘤在BALB/c裸鼠体内生长情况的影响。Westernblot检测瘤组织中增殖细胞核抗原(proliferatingcellnuclearantigen,PCNA)的表达情况。结果:30mg/LDADS处理的MGC803细胞移植裸鼠体内无一成瘤。荷瘤裸鼠腹腔注射DADS剂量为50、100和200mg/kg时的抑瘤率分别为27.8%、66.1%和73.0%,同时可抑制移植瘤癌细胞PCNA的表达。结论:DADS可明显降低胃癌细胞裸鼠移植瘤的成瘤性,并对移植瘤生长有明显抑制作用。     

早期和延期腹腔内化疗对结肠癌细胞腹腔种植的影响

采用人结肠癌裸鼠腹腔种植瘤模型，进行表阿霉素早期和延期腹腔内化疗，观察其对癌细胞腹腔内种植的影响。15只雄性裸鼠腹腔内接种Lovo细胞后，随机分成对照组、早期和延期腹腔内化疗3组。早期腹腔内化疗组以0．8mg/kg/d表阿霉素于接种后1～3天行腹腔注射，延期组用相同剂量于接种后5～7天行腹腔注射。结果显示，早期腹腔内化疗组裸鼠平均荷瘤生存时间最长；延期腹腔内化疗组次之；对照组最短。前者与后两者比较，均有显著性差异（P＜0．05和P＜0．01）。实验提示早期腹腔内化疗的抑瘤作用明显高于延期腹腔内化疗。     

In vivo metastasis of human tumor transplant in T and NK cell immune-deficient mice

Beige-nude mice with combined T and NK cell deficiency were produced by introducing "nu" gene into the beige mice of C57BL/6 background. The beige-nude mice were characterized by morphological features, level and activity of the NK cell in spleen and peripheral blood. NK cell level of the beige-nude mice was higher than the beige mice and similar to the ordinary nude mice though its NK cell activity was markedly lower than the nude mice. Human lung adenocarcinoma (PAa) was transplanted subcutaneously to the beige-nude mice and nude mice of BALB/cA background. No metastasis was found in BALB/cA nude mice (0/7), while in beige-nude mice, spontaneous lymph node metastasis rate was 36.3% (4/11). It should be noted that the transplanted PAa metastasized to lungs, which had never been seen in BALB/cA nude mice in previous experiments. The increase of metastatic rate is correlated with the relative low NK cell activity of the beige-nude mice. Our observation is that the beige-nude mice could be a suitable experimental model for in vivo study on metastatic behavior of the human tumors.     

Steroid hormones stimulate human prostate cancer progression and metastasis

Tissue recombinants (TRs) composed of mouse urogenital mesenchyme (mUGM) plus an immortalized nontumorigenic human prostatic epithelial cell line (BPH-1) were grown under the kidney capsule of male athymic nude mice under different hormonal conditions. The objectives were to determine temporal plasma concentrations of testosterone (T) and estradiol-17beta (E2) that elicit progression of nontumorigenic human prostatic epithelial cells in vivo. Second, to determine whether mUGM+BPH-1 TRs in [T+E2]-treated hosts could progress to metastases. Control mouse hosts received no exogenous hormonal support, whereas treated mice received Silastic implants containing T and E2 for 1-4 months. Plasma from hormonally treated mice contained significantly higher (p < 0.01) concentrations of T at 1 month (11.7 vs. 0.9 ng/ml). Plasma levels of E2 in steroid implanted mice were significantly higher (p < 0.05) at 2 months (104.5 vs. 25.6 ng/l) and 4 months (122.8 vs. 19.2 pg/ml). Wet weights of mUGM+BPH-1 TRs from [T+E2]-implanted mice were significantly larger (p < 0.001) than those from untreated hosts. Untreated mUGM+BPH-1 TRs contained a well organized differentiated epithelium surrounded by smooth muscle stroma similar to developing prostate. In [T+E2]-implanted mice, mUGM+BPH-1 TRs formed carcinomas that contained a fibrous connective tissue stroma permeating the tumor; smooth muscle when present was associated with vasculature. Renal lymph nodes collected from [T+E2]-treated mice, but not untreated mice, contained metastatic carcinoma cells. Moreover, metastases could be observed at distant sites including lung and liver. Epithelial cells isolated from untreated mUGM+BPH-1 TRs exhibited benign histology and formed small nontumorigenic grafts when subsequently transplanted into athymic nude mice. In contrast, epithelial cells isolated from mUGM+BPH-1 tumors of [T+E2]-treated hosts formed large tumors that grew independent of stromal and hormonal support and developed lymph node metastases. We conclude that [T+E2]-treatment promotes prostatic cancer progression in mUGM+BPH-1 TRs. Use of mUGM in this system will allow future studies to utilize the power of mouse genetics to identify paracrine factors involved in human prostatic carcinogenesis.