人类胃癌裸鼠淋巴结转移模型的建立

 目的　研究胃癌淋巴结转移动物模型的建立方法。方法　人类胃癌低分化细胞系SGC-7901体外培养、传代并扩增后，收集细胞行皮下种植成瘤，鼠间传代至第6代，以皮下肿瘤组织块原位种植于裸鼠胃壁建立动物模型。种植后第9周处死裸鼠，观察原位种植瘤生长、淋巴结转移及其他脏器转移情况，测定荷瘤裸鼠血清癌胚抗原（CEA）值。结果　原位移植瘤种植成功率100 ％，胃周淋巴结转移率93.3 ％，移植瘤可发生局部浸润及远处脏器转移，荷瘤裸鼠CEA值明显高于正常裸鼠（P＜0.01）。结论　应用SGC-7901细胞系可成功建立胃癌的淋巴结转移动物模型。     

人类卵巢癌原位移植自发转移裸小鼠模型的构建

 目的　用人类卵巢癌组织块原位移植构建与人类卵巢癌最相似的裸鼠原位移植自发转移模型。方法　将人类卵巢癌高转移细胞系8910PM在裸鼠皮下注射成瘤,然后将瘤组织运用显微外科的方法植入裸鼠卵巢包膜内,原位移植成功后继续在鼠间卵巢包膜内传代,并运用透射电镜进一步验证肿瘤形成和转移效果。结果　裸鼠皮下与卵巢部位成瘤率均为100 %（分别为2／2和16／16）,原位移植瘤转移率为50 %。卵巢包膜间传代,第2代、第4代分别出现卵巢癌的肝转移,其他部位尚未见转移灶。最早成瘤时间1周,最早转移出现时间2周。在连续传代过程中移植瘤仍保持原癌组织病理形态特点及人类肿瘤染色体的特点。结论　成功构建了人类卵巢癌原位移植自发转移裸鼠模型,为卵巢癌的转移机制及治疗研究提供了一个理想的动物模型。     

Establishment of a human chorionic gonadotropin-producing human gastric carcinoma in nude mice

A human chorionic gonadotropin (HCG)-producing gastric carcinoma was transplanted into BALB/c nu/nu nude mice. The original tumor tissue had been obtained by gastrectomy from a 55-year-old patient with gastric carcinoma. The tumor was transplanted serially in nude mice, and its doubling time was stable to approximately 12 days. A positive correlation was observed between serum HCG level and tumor weight. The serum HCG level of the mice and the histology of the tumor weight. The serum HCG level of the mice and the histology of the tumor, papillary adenocarcinoma with partial poorly differentiated adenocarcinoma, did not change on serial transfers. HCG was positively stained by an immunochemical technique only in the site of the poorly differentiated adenocarcinoma, while staining was negative in the site of the papillary adenocarcinoma, which constituted most of the tumor. These results indicate that this tumor in nude mice will be useful not only as a therapeutic experimental system but also for studying the potential malignancy of HCG-producing cancer cells.     

Invasion and metastasis of SY86B human gastric carcinoma cells in nude mice

A moderately differentiated tubular adenocarcinoma of human stomach, named SY86B, was successfully transplanted subcutaneously to nude mice of different genetic backgrounds (BALB/CA/PBI-nu, C57BL/6J.615/PBI-nu and ICR.BALB/CA/PBI-nu). The tumor has been passaged for 13 generations and the transplantability was 100%. The SY86B cells retained the capacity of invasive and metastatic growth in the nude mice and showed a high rate of metastasis to the regional lymph nodes and to such distant organs as the lungs, liver and pancreas. The overall rate of metastasis was 77.7%. The species of the nude mice, their age and sex apparently did not significantly affect the occurrence of metastasis. Tumor-bearing time and the aggressive character of the tumor cells themselves appeared important for the genesis of metastasis. This experimental model can provide a new approach to basic and clinical studies of cancer metastasis.     

Establishment and characterization of primary human pancreatic carcinoma in continuous cell culture and in nude mice.

Primary human panceratic exocrine adenocarcinoma has been established in tissue culture and as xenografts in immune-deficient nu/nu mice. The cell line has a doubling time of 36 h and grows as a confluent monolayer together with a constant population of free-floating cells. Evidence of tumourigenicity was provided by growth on an early diploid fibroblast monolayer and in soft agar, and as solid tumours in immune-deficient nu/mu mice. Chromosome analysis of the cultured cells confirmed their tumour origin. Xenografts established from the cell line or directly from primary tumour tissue have retained a similar histology to the original tumour on serial transplantation. An electrophoretic study of exportable pancreatic digestive enzymes and a number of intracellular enzymes has shown that the cell line and xenografts maintain a human intracellular enzyme profile, but do not produce pancreatic digestive enzymes.     

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.     

高表达MUC1的人食管癌裸鼠移植瘤模型的建立

背景与目的:Mucin-1(MUC1)粘蛋白是一种肿瘤相关抗原,是肿瘤免疫治疗良好的分子靶点之一.本研究建立高表达MUCl的人食管癌裸鼠皮下移植瘤模型,为研究以MUC1为靶点的食管癌的生物治疗提供体内模型.方法:以体外培养的高表达MUC1的人食管癌细胞株EC-109接种于4～5周龄的BALB/c裸小鼠皮下,观察移植瘤生长情况,对移植瘤进行病理组织学检查,采用免疫组化方法检测移植瘤细胞增殖细胞核抗原(proliferating cell nuclear antigens,PCNA),采用流式细胞仪检测移植瘤细胞的细胞周期及MUC1的表达.结果:裸鼠皮下移植瘤成瘤率为86.0%,移植瘤具有与人恶性肿瘤相似的组织学和生物学特点,其平均PCNA标记指数为(63.5 3.6)%、S期细胞百分比(S-phase fraction,SPF)平均值为(37.6±3.7)%、MUC1的平均表达率为97.5%.结论:高表达MUC1的人食管癌裸鼠皮下移植瘤模型具有恶性肿瘤的生物学特性,可用于研究人食管癌的生物学特点,同时为以MUC1为靶点的食管癌的免疫治疗研究提供了良好的体内模型.     

Tumorigenicity and metastasis of human breast carcinoma cell lines in nude mice

There are few reports describing experimental models of the growth and metastasis of human breast carcinomas. This article discusses the tumorigenic and metastatic properties of two estrogen receptor-negative breast carcinomas injected into nude mice. Tumor growth in the mammary fatpad (m.f.p.) and the subcutis was compared in female nude mice. The injection of 10(5) viable cells of two human breast carcinoma cell lines (MDA-MB-231 and MDA-MB-435) gave a 100% tumor take rate in the m.f.p., whereas only 40% of the s.c. injections produced tumors and these occurred several weeks after the appearance of the m.f.p. tumors. Thus, the m.f.p. of nude mice is a favorable site for the growth of human breast carcinomas. MDA-MB-435 tumors produced distant metastases in 80% to 100% of recipients. The most common sites for metastasis were the lymph nodes and lungs, with a lower incidence of metastases in muscle (chest wall and thigh), heart, and brain. New variant cell lines were isolated from metastases in the lungs, brain, and heart. All the cell lines were tumorigenic in the m.f.p., and the lung- and heart-derived metastasis lines produced slightly more lung metastases than the original cell line. However, the brain metastasis variant produced significantly fewer lung metastases. Intravenous inoculation of the spontaneous metastasis-derived cell lines produced few lung colonies. Only cell variants isolated from experimental lung metastases showed enhanced lung colonization potential when reinjected i.v. Our results suggest that the estrogen receptor-negative MDA-MB-435 cell line injected in the m.f.p. of nude mice could be a valuable tool for analysis of the cellular and molecular basis of the metastasis of advanced breast cancer.     

Establishment and characterisation of a human clear cell sarcoma model in nude mice

We have established a new experimental model of human clear cell sarcoma, UM-CCS1, using serial subcutaneous transplantation of intact tumour tissue in nude mice. The heterotransplanted nude mouse tumours retained characteristic morphological features of the primary clear cell sarcoma. Immunohistochemical analysis showed the retained expression patterns of S-100 protein, melanoma-associated antigen HMB-45 and vimentin in the xenografts as compared to the primary tumour. DNA index showed low variations both between the xenografts in the same passage and between the serial passages. Cytogenetic analysis of the primary tumour and the xenografts showed the unbalanced translocation der(6)t(6;12)(p23;q13). Based on the combined genetic data a reasonable interpretation of our findings is that there was a complex chromosomal rearrangement resulting in a cytogenetically cryptic EWS-ATF1 fusion gene. Analysis of cell kinetics using in vivo incorporation of iododeoxyuridine and flow cytometry showed generally short potential doubling time (T(pot)) of the xenografts. Volume doubling time showed low variations without correlation with T(pot). The retained phenotypic and genotypic characteristics of the primary tumour and the morphological and structural stability over time makes the model suitable for studies on the tumour biology and treatment of clear cell sarcoma.     

人子宫内膜癌瘤组织块活体移植瘤动物模型的建立

目的:研究建立人子宫内膜癌的组织块活体移植动物模型方法。方法:将生长状态良好的Ishikawa细胞浓度调整至1.5×107/ml,接种于BALB/c nude裸鼠腋下皮下(Ⅰ组),Ⅱ组生理盐水对照,观察其生长情况,待成瘤5-10mm时建立成瘤模型,收集Ⅰ组瘤组织块行活体移植于Ⅲ组小鼠,留取组织标本行病理检查。结果:成功建立了人子宫内膜癌Ishikawa细胞的BABL/c裸鼠皮下移植瘤模型和活体移植瘤模型,形成的肿瘤具备人肿瘤生物学特点。结论:建立的瘤组织块活体二次传代移植人子宫内膜癌的BABL/c nude小鼠皮下移植瘤模型具有人子宫内膜癌特征且成瘤率更高,为大批建立子宫内膜癌单位模型研究子宫内膜癌的发病机制和药物治疗提供了实验动物模型。     

不同转移潜能MG63骨肉瘤细胞亚株裸鼠肺转移模型的建立

Objective: To establish a nude mice model of human osteosarcoma lung metastasis. Methods: The growth of human osteosarcoma cell sublines M8 and M6 was determined by MTT assay. 2 × 107 cells were injected into the tail vein of nude mice. Mice were sacrificed started on week 4 after injection, and lung metastases were evaluated under both mac-roscopic and microscopic observation with HE staining. Results: The growth of low-metastatic subline M6 was lower than high-metastatic sublines M8. Seventeen mice after injected M8 had occurred lung metastases while only one mice had oc-curred in M6 group. Moreover, M8 cells within metastases were arrangement disorder with variable nuclear hyperchromasia. Conclusion: A mouse model for human osteosarcoma cancer lung metastasis can be established by injection different ability of metastasis MG63 cells into tail vein.     

Establishment and characterization of human uveal malignant melanoma xenografts in nude mice

The purpose of this study was to develop a suitable animal model for the investigation of the pathogenesis and therapy of uveal malignant melanoma. Eight choroidal malignant melanomas from eight patients were transplanted into nude mice in an attempt to establish a serially transplantable tumour model. Tumour tissue blocks (2 x 2 x 2 mm) from enucleated eyes with choroidal malignant melanoma were transplanted subcutaneously into the flanks of nude mice. The growing tumours were measured and serially transplanted. The tumour samples were investigated by histology, immunohistochemistry and electron microscopy. Only one of the eight transplanted primary tumours (13%) was established as a xenograft in nude mice. Furthermore, the take rate of the transplantable tumour was low (13%). The growth of the tumour fitted a Gompertz function, and the calculated tumour volume doubling time was 54 days. The transplanted tumour cells were epithelioid and slightly larger than the primary tumour cells and had prominent nucleoli. However, the transplanted tumour retained a morphological appearance similar to that of the primary tumour. Immunohistochemical examinations demonstrated that the cells preserved the characteristic properties of malignant melanoma. However, the transplanted cells demonstrated vimentin reactivity, whereas the primary tumour cells were negative for vimentin. It can be concluded that a new experimental model of malignant uveal melanoma with tumours that were easy to observe and access was established in nude mice.     

Transplantation model of human hepatocellular carcinoma in nude mice. II. Establishment of the LTNM2 human liver cancer model in nude mice and observation on the growth of the transplanted tumor

LTNM2 model was established by the surgical specimen of a patient with recurrent hepatocellular carcinoma (HCC) with positive AFP. Human HCC was successfully transplanted subcutaneously into nude mice (Swiss nu/nu) following a latent period of 93 days with serial passages for 7 generations. Transplantability in nude mice was 72% (42/58). Based on the observation of 42 nude mice models, it was proved that the original properties of the human HCC, such as the morphological features, cell type and degree of differentiation and the functions, synthesis of AFP, were preserved. The tumor growth was progressive. The mean value of increased geometrical mean diameter tumor growth was progressive. The mean value of increased geometrical mean diameter (GMD) was 2.5 +/- 1.1 mm/week. Necrosis, ulceration and spontaneous regression was rarely observed. These results show that the transplanted tumor in nude mice bears a strong resemblance to the parent human HCC. LTNM2 model can be used not only for basic research of liver cancer but also for experimental therapeutic study.     

CIK细胞对裸鼠腹膜移植人胃癌细胞增殖和凋亡影响的实验研究

目的研究细胞因子诱导的杀伤细胞(Cytokine inducedkiller,CIK)对裸鼠腹膜移植人胃癌细胞增殖和凋亡的影响。方法体外制备CIK细胞。建立人胃癌细胞裸鼠腹膜移植小鼠模型,模型鼠腹腔注射CIK细胞,透射电镜观察腹膜移植癌组织形态学变化,流式细胞术(FCM)检测癌细胞增殖周期中G0/G1、S、G2/M期细胞比率、增殖指数(proliferationIndex,PI)和细胞凋亡指数(apoptosisIndex,AI)的变化。结果模型鼠腹膜癌组织细胞的超微结构出现不同程度的凋亡形态学改变,FCM检测DNA直方图上呈现有凋亡峰。G0/G1期细胞比率为(70.73±2.02)%～(75.88±2.23)%,AI为(7.50±2.69)%～(13.16±2.58)%,与对照组相比显著增加,P=0.0000,P=0.008,P<0.01;S期细胞比率为(14.96±1.32)%～(15.34±1.30)%,无显著变化,P>0.05;G2/M期细胞比率为(14.31±1.51)%～(8.91±0.86)%,PI为(29.27±2.05)%～(24.12±2.10)%,均显著降低,P=0.000、0.008、0.033、0.043,P<0.01。随CIK细胞注射剂量的增加和作用时间延长,G0/G1期细胞比率和AI逐渐增加,而G2/M期细胞比率和PI下降,差异均有统计学意义,P<0.05或P<0.01。细胞AI与G0/G1期细胞比率正相关,r=0.676,与PI负相关,r=-0.659。结论CIK细胞可以影响胃癌腹膜种植转移细胞的细胞周期,并具有诱导细胞凋亡的作用。     

A new experimental metastasis model in athymic nude mice, the human malignant melanoma LOX

The human tumor line LOX was established as an s.c. xenograft in nude mice from a lymph-node metastasis of a patient with malignant melanoma. I.v. injection into adult nude mice of single-cell suspensions prepared from xenografts resulted in progressively growing lung tumor colonies that killed the animals. No difference in colony formation was seen between cells taken from lung colonies and s.c. xenografts. An in vitro cell line, LOX-L, was established from lung colonies, and the monolayer cells, detached with EDTA, retained the same ability to form experimental lung metastases. In a total of 14 experiments, 82 of 89 mice receiving 1 X 10(6) viable tumor cells died with a mean survival time of 34.1 +/- 4.8 days. Long-term passaging in vivo and in vitro did not result in any alteration of the lung-colonizing potential of the LOX cells, whereas trypsinization of the cells before i.v. injection reduced lung colony formation. The life span was inversely related to the number of LOX cells injected, permitting estimation of the cell kill caused by chemotherapy. Mice injected i.v. with the LOX cells showed the same relative response to cis-diamminedichloroplatinum (CDDP) and mitozolomide (MZA) as did animals carrying s.c. xenografts. The LOX cells have shown a remarkable stability and similarity to the cells of the patient's tumor with respect to morphology, karyotype and chemosensitivity. The LOX model may be useful for testing effects of therapy on lung micro- and macrometastases, and the activity of antimetastatic agents, as well as for studying mechanisms involved in the metastatic process.     

Establishment and characterization of human gastric carcinoma cell lines

We report 8 newly established gastric-carcinoma cell lines (SNU-216, 484, 520, 601, 620, 638, 668, 719) from Korean patients. Morphologic study was carried out using light and electron microscopes. CEA, αFP, and CA 19-9 and TPA in supernatant and in cell lysate were measured by radioimmunoassay. p53 and c-Ki - ras gene mutations were screened and confirmed by sequencing. The cell lines, derived from tumors with moderate differentiation, grew as a diffuse monolayer, and those from tumors with poor differentiation and minimal desmoplasia grew exclusively as non-adherent. Out of the 8 gastric-cancer cell lines, 5 had detectable levels of CEA both in supernatant and in cell lysate; there was no expression or secretion of αFP in these cells; 4 cell lines showed high levels of CA 19-9 in cell pellets. All cell lines except SNU-484 had high concentrations of TPA both in cell lysate and in supernatants. p53 mutation was found in 6 cell lines (75%): 2 (SNU-216 and SNU-668) had mutations in exon 6, and other 3 in exon 8. The c-Ki - ras mutation was found in 2 cell lines (25%), SNU-601 and SNU-668. The former showed GGT-to-GAT transition mutation at codon 12, while the latter showed CAA-to-AAA transversion mutation at codon 61. DNA profiles using restriction endonuclease Hinfl and polymorphic DNA probes ChdTC-15 and ChdTC-114 showed different unique patterns; which suggests that these cell lines are unique and not cross-contaminated. We believe that the newly characterized gastric-cancer cell lines presented in this paper will provide a useful in vitro model for studies related to human gastric cancer. Int. J. Cancer, 70:0–0, 1997. © 1997 Wiley-Liss, Inc.     

人胃癌细胞株BGC-823在裸鼠腹腔内种植转移后大网膜巨噬细胞的演化

目的:探讨人胃癌细胞种植到大网膜后巨噬细胞的动态表达和形态改变及其意义。方法:采用动物模型制备人胃癌转移后的裸鼠大网膜标本,免疫组织化学法检测80例大网膜标本中巨噬细胞F4/80的表达情况,透射电子显微镜观察巨噬细胞超微结构变化。结果:实验组和对照组大网膜巨噬细胞表达率和形态改变存在差异。自第3天始,实验组和对照组巨噬细胞表达率即出现差异,实验组巨噬细胞表达强;实验组不同种植时间巨噬细胞表达率有差异,对照组则否。实验组大网膜巨噬细胞F4/80表达率随种植时间的延长不断增强,两者呈正相关(P<0.001),对照组大网膜巨噬细胞F4/80表达率不随种植时间的延长而增强;实验组大网膜巨噬细胞较对照组变化多。结论:大网膜巨噬细胞数量增多和形态改变最终作用是为胃癌细胞的种植转移提供良好的微环境条件。     

p53 expression,growth, and spontaneous metastasis of the human GI 101 breast carcinoma in athymic nude mice

The human GI 101 breast carcinoma cell lines produces spontaneous metastasis to the lungs when xenografted subcutaneously in female athymic nude mice. To establish the time-course of tumor growth and distant metastasis to the lungs and axillary lymph nodes, 5 mm3 of tumor tissue was implanted in the subaxial region of female athymic nude mice. Micrometastases in the lung were first detected 3 weeks after tumor implantation. The incidence of lung metastasis and the number of tumor emboli were correlated with the volume of the primary tumors. Ipsilateral axillary lymph node metastasis was observed within 17 weeks, indicating that metastasis to the lymph node is a later event. Unlike pulmonary micrometastases which were in the form of clusters of four to six tumor cells, metastasis to the lymph nodes were in nodules of poorly differentiated and larger tumor cells. Immunohistochemistry evaluation of p53 oncoprotein in the primary and metastatic tumor cells showed different patterns of subcellular accumulation. Cytoplasmic staining was mainly detected in the primary and secondary tumor cells disseminated to the lungs. In contrast, nuclear staining was only detected in tumor cells infiltrated to the axillary lymph nodes. There was no gain of loss of positivity of p53 accumulation (i.e., qualitative measurements) as the tumor grew in size. The data indicate that the GI 101 tumor cells could be used as a useful model for studying the malignant progression of hormone-independent breast cancer, antimetastatic drugs, and early events in tumor metastasis.     

Establishment of a human colonic carcinoma xenograft in nude mice and its chief biological characteristics

A human colonic cancer specimen cut into 1-2 mm3 pieces under aseptic condition was heterotransplanted hypodermically by trocar to the inguinal region of BALB/cATcL nude mice bred in our laboratory. 53 days later, a 10 X 9 X 11 mm3 tumor was obtained. The take rate was 100% (48/48). The take rate of liquid nitrogen frozen and recovered tissue pieces was also 100% (13/13). Grossly, the tumor was rich in blood supply and well encapsulated. Within the tumor, the tissue was cream-colored. G-banded chromosome analysis revealed a human chromosome pattern with the mode 70-90. Pathologic diagnosis was signet-ring cell cancer, which was identical to the surgical specimen. No change was found after maintaining 13 passages in nude mice. It was named the human colonic carcinoma xenograft XHCn/w. It provides an ideal tumor model for in vivo or in vitro experiments.     

康莱特注射液对结肠癌肝转移裸鼠血浆骨桥蛋白的影响

目的　建立人结肠癌ＬｏＶｏ细胞裸鼠肝转移模型,观察康莱特注射液（ＫＬＴ）对该模型裸鼠血浆骨桥蛋白的影响。方法　将３０只裸鼠随机分为５组：对照组、环磷酰胺（ＣＴＸ）组、ＫＬＴ组、ＣＴＸ＋ＫＬＴ组和正常组。前４组将０.２ｍｌＬｏＶｏ细胞悬液（１×１０７个／ｍｌ）接种于裸鼠脾脏,建立结肠癌肝转移模型,分别经腹腔注射给予生理盐水、ＣＴＸ、ＫＬＴ和ＣＴＸ＋ＫＬＴ。正常组未接种肿瘤细胞。接种４５天后处死全部裸鼠,称取４组脾脏接种瘤和肝脏转移瘤重量,并检测血浆骨桥蛋白浓度。结果　与对照组相比,ＣＴＸ组、ＫＬＴ组、ＣＴＸ＋ＫＬＴ组的脾脏接种瘤和肝脏转移瘤重量均减轻（Ｐ＜０.０５）,其中ＫＬＴ组分别为（４０１.７±５３.１）ｍｇ和（３５３.３±３９.８）ｍｇ,ＣＴＸ组＋ＫＬＴ组分别为（２３５.０±５２.４）ｍｇ和（１８５.０±２７.４）ｍｇ。与正常组相比,其余４组裸鼠血浆骨桥蛋白浓度均升高（Ｐ＜０.０５）;对照组血浆骨桥蛋白浓度为（２.１０±０.５５）ｎｇ／ｍｌ,ＣＴＸ＋ＫＬＴ组为（０.９２±０.３１）ｎｇ／ｍｌ,两组差异有统计学意义（Ｐ＜０.０５）,而ＣＴＸ组、ＫＬＴ组与对照组比较差异无统计学意义（Ｐ＞０.０５）。结论　ＫＬＴ联合ＣＴＸ可以抑制模型裸鼠的脾脏接种瘤及肝脏转移瘤的生长,抑制其血浆骨桥蛋白的浓度。