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1.
Rashidi B Yang M Jiang P Baranov E An Z Wang X Moossa AR Hoffman RM 《Clinical & experimental metastasis》2000,18(1):57-60
The Lewis lung carcinoma has been widely used for many important studies. However, the subcutaneous transplant or orthotopic cell-suspension injection models have not allowed the expression of its full metastatic potential. A powerful new highly metastatic model of the widely-used Lewis lung carcinoma is reported here using surgical orthotopic implantation (SOI) of tumor fragments and enhanced green fluorescent protein (GFP) transduction of the tumor cells. To achieve this goal, we first developed in vitro a stable high-expression GFP transductant of the Lewis lung carcinoma with the pLEIN retroviral expression vector containing the enhanced Aequorea victoria GFP gene. Stable high-level expression of GFP was found maintained in vivo in subcutaneously-growing Lewis lung tumors. The in vivo GFP-expressing tumors were harvested and implanted as tissue fragments by SOI in the right lung of additional nude mice. This model resulted in rapid orthotopic growth and extensive metastasis visualized by GFP-expression. 100% of the animals had metastases on the ipsilateral diaphragmatic surface, contralateral diaphragmatic surface, contralateral lung parenchima, and in mediastinal lymph nodes. Heart metastases were visualized in 40%, and brain metastases were visualized in 30% of the SOI animals. Mice developed signs of respiratory distress between 10–15 days post-tumor implantation and were sacrificed. The use of GFP-transduced Lewis lung carcinoma transplanted by SOI reveals for the first time the high malignancy of this tumor and provides an important useful model for metastasis, angiogenesis and therapeutic studies. 相似文献
2.
Yang M Chishima T Wang X Baranov E Shimada H Moossa AR Hoffman RM 《Clinical & experimental metastasis》1999,17(5):417-422
Stable high-level green fluorescent protein (GFP)-expressing Chinese hamster ovary cells (CHO) were used to visualize the degree of metastatic behavior of this cell line in nude and SCID mice. A stable GFP high-expression CHO clone, selected in 1.5 M methotrexate, was injected subcutaneously in nude and severe combined immunodeficient (SCID) mice and implanted orthotopically in the ovary of nude mice. CHO proved to be highly metastatic from both the subcutaneous and orthotopic sites as brightly visualized by GFP fluorescence. High-level GFP-expression allowed the visualization of metastatic tumor in fresh live host tissue in great detail. Metastases were visualized by GFP expression in the lung, pleural membrane, spleen, kidney, ovary, adrenal gland, and peritoneum after orthotopic implantation in nude mice. Metastases were visualized by GFP expression mainly in the lung, pleural membrane after subcutaneous implantation in nude mice. Metastases were visualized in the lung and pleural membrane, liver, kidney, and ovary after subcutaneous implantation in SCID mice. The construction of highly fluorescent stable GFP transfectants of CHO has revealed the multi-organ metastatic capability of CHO cells. CHO has such a high degree of malignancy that it is metastatic from both the orthotopic and subcutaneous transplant sites. This highly malignant GFP-expressing cell-line with multi-organ metastatic affinity should serve as a powerful tool to study tumor-host interaction. 相似文献
3.
Imaging tumor angiogenesis with fluorescent proteins 总被引:5,自引:0,他引:5
Hoffman RM 《APMIS : acta pathologica, microbiologica, et immunologica Scandinavica》2004,112(7-8):441-449
We have developed three unique mouse models to image angiogenesis with fluorescent proteins, which are described in this review. First, we have adapted the surgical orthotopic implantation (SOI) model to image angiogenesis of human tumors labeled with green fluorescent protein (GFP) transplanted in nude mice. The nonluminous induced capillaries are clearly visible by contrast against the very bright tumor fluorescence examined either intravitally or by whole-body imaging in real time. Intravital images of an SOI model of human pancreatic tumors expressing GFP visualized angiogenic capillaries at both primary and metastatic sites. Whole-body optical imaging showed that blood vessel density increased linearly over a 20-week period in an SOI model of human breast cancer expressing GFP. Opening a reversible skin-flap in the light path markedly reduces signal attenuation, increasing detection sensitivity many-fold and enabling vessels to be externally visualized in GFP-expressing tumors growing on internal organs. The second model utilizes dual-color fluorescence imaging, effected by using red fluorescent protein (RFP)-expressing tumors growing in GFP-expressing transgenic mice that express GFP in all cells. This dual-color model visualizes with great clarity the details of the tumor-stroma interaction, especially tumor-induced angiogenesis. The GFP-expressing tumor vasculature, both nascent and mature, are readily distinguished interacting with the RFP-expressing tumor cells. Using a spectral imaging system based on liquid crystal tunable filters, we were able to separate individual spectral species on a pixel-by-pixel basis. Such techniques non-invasively visualized the presence of host GFP-expressing vessels within an RFP-labeled orthotopic human breast tumor by real-time whole-body imaging. The third model involves a transgenic mouse in which the regulatory elements of the stem cell marker nestin drive GFP. The nestin-GFP mouse expresses GFP in areas of the brain, hair follicle stem cells, and in a network of blood vessels in the skin interconnecting hair follicles. RFP-expressing tumors transplanted to nestin-GFP mice enable specific visualization of nascent vessels in skin-growing tumors such as melanoma. Thus, fluorescent proteins expressed in vivo offer very high resolution and sensitivity for real-time imaging of angiogenesis. 相似文献
4.
Zili An Ping Jiang Xiaoen Wang A.R. Moossa Robert M. Hoffman 《Clinical & experimental metastasis》1999,17(3):265-270
In this study we compared the metastatic rate of human renal cell carcinoma SN12C in two orthotopic nude mouse models: surgical orthotopic implantation (SOI) of histologically intact tumor tissue and cellular orthotopic injection (COI) of cell suspensions in the kidney. The primary tumors resulting from SOI were larger and much more locally invasive than primary tumors resulting from COI. SOI generated higher metastatic expression than COI. The differences in metastatic rates in the involved organs (lung, liver, and mediastinal lymph nodes) were 2–3 fold higher in SOI compared to COI (P<0.05). Median survival time in the SOI model was 40 days, which was significantly shorter than that of COI (68 days) (P<0.001). Histological observation of the primary tumors from the SOI model demonstrated a much richer vascular network than the COI model. Lymph node and lung metastases were larger and more cellular in the SOI model compared to COI. We conclude that the tissue architecture of the implanted tumor tissue in the SOI model plays an important role in the initiation of primary tumor growth, invasion, and distant metastasis. This study directly demonstrates that the implantation of histologically intact tumor tissue orthotopically allows accurate expression of the clinical features of human renal cancer in nude mice. This model should be of value in cancer research and antimetastatic drug discovery for renal cancer, a currently very poorly responding malignancy. 相似文献
5.
Li X Wang J An Z Yang M Baranov E Jiang P Sun F Moossa AR Hoffman RM 《Clinical & experimental metastasis》2002,19(4):347-350
We report an optically imageable orthotopic metastatic nude mouse model of the human breast cancer MDA-MB-435 expressing green
fluorescent protein (GFP). We demonstrate fluorescent imaging of primary and metastatic growth in live tissue and in intact
animals. Fragments of tumor tissue expressing GFP were sutured into the pocket in the right second mammary gland. Tumor tissue
was strongly fluorescent, enabling whole-body imaging of tumor growth by week 5. Neovascularization of the primary tumor was
also visualized by whole-body imaging by contrast of the vessels to the fluorescent tumor. At autopsy, the MDA-MB-435-GFP
was found to have metastasized to various organs, including the lung in 55% of the animals, the lymph nodes in 15% of the
animals including axillary nodes, and the liver in 10% of the animals. These metastases could be visualized in fresh tissue
by fluorescent imaging. Detailed fluorescence analysis visualized extensive metastasis in the thoracic cavity and the lymphatic
system. Large metastatic nodules in the lung involved most of the pulmonary parenchyma in all lobes. Lymph node metastasis
was found mainly in the axillary area. In the liver, fluorescent macroscopic metastatic nodules were found under the capsule.
The metastatic pattern in the model thus reflected clinical metastatic breast cancer and provides a powerful model for drug
discovery for this disease.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
6.
Lee NC Bouvet M Nardin S Jiang P Baranov E Rashidi B Yang M Wang X Moossa AR Hoffma RM 《Clinical & experimental metastasis》2000,18(5):379-384
Gemcitabine is a promising new agent that has been recently studied for palliation of advanced (stage IV) unresectable pancreatic
cancer. We hypothesized that adjuvant gemcitabine would reduce recurrence and metastases following surgical resection of pancreatic
cancer. To test this hypothesis, we evaluated gemcitabine on a green fluorescent protein (GFP) transductant of the human pancreatic
cancer cell line BxPC-3 (BxPC-3-GFP) using surgical orthotopic implantation (SOI) in nude mice. GFP enabled high resolution
fluorescent visualization of primary and metastatic growth. Five weeks after SOI, the mice were randomized into three groups:
Group I received exploratory laparotomy only. Group II underwent surgical resection of the pancreatic tumor without further
treatment. Group III underwent tumor resection followed by adjuvant treatment with gemcitabine, 100 mg/kg every three days
for a total of four doses, starting two days after resection. The mice were sacrificed at thirteen weeks following implantation
and the presence and location of recurrent tumor was recorded. Gemcitabine reduced the recurrence rate to 28.6% compared to
70.6% with resection only (P=0.02) and reduced metastatic events 58% in the adjuvant group compared to resection only. This study, demonstrating that
gemcitabine is effective as adjuvant chemotherapy post-pancreatectomy, suggests this new indication of the drug clinically.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
7.
Bouvet M Yang M Nardin S Wang X Jiang P Baranov E Moossa AR Hoffman RM 《Clinical & experimental metastasis》2000,18(3):213-218
Pancreatic cancer is a highly metastatic disease that responds poorly to currently-available treatment. In order to better
visualize and understand the chronology and specificity of metastatic targeting of pancreatic cancer, two human pancreatic
cancer cell lines, expressing green fluorescent protein (GFP), were studied in orthotopic models. MIA-PaCa2-GFP and BxPC-3-GFP
tumor fragments were transplanted by surgical orthotopic implantation (SOI) to the nude mouse pancreas for fluorescence visualization
of the chronology of pancreatic tumor growth and metastatic targeting. BxPC-3-GFP tumors developed rapidly in the pancreas
and spread regionally to the spleen and retroperitoneum as early as six weeks. Distant metastases in BxPC-3-GFP were rare.
In contrast, MIA-PaCa-2-GFP grew more slowly in the pancreas but rapidly metastasized to distant sites including liver and
portal lymph nodes. Regional metastases in MIA-PaCa-2-GFP were rare. These studies demonstrate that pancreatic cancers have
highly specific and individual “seed-soil” interactions governing the chronology and sites of metastatic targeting.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
8.
Bobek V Kolostova K Pinterov D Boubelik M Jiang P Yang M Hoffman RM 《Clinical & experimental metastasis》2004,21(8):705-708
The Lewis lung tumor has been extensively studied in both syngeneic and allogeneic mouse models. However, its metastatic potential and mechanism are poorly understood. The aim of the present study was to develop a highly metastatic lymph-node targeting, imageable model of the Lewis lung carcinoma in a syngeneic host. We report here a syngeneic model of the Lewis lung carcinoma in which the carcinoma cells are labeled with green fluorescent protein (GFP). The tumor cells were transplanted in the dorsal side of the ear of C57-B16 mice in order to give the tumor cells access to the lymphatic system. This model of the Lewis lung carcinoma extensively metastasized to numerous lymph nodes throughout the body of the animal as well as visceral organs, as visualized by fluorescence microscopy using the bright GFP signal. Twenty-one different metastatic sites, including lymph nodes throughout the body, were identified among the cohort of transplanted animals. The data demonstrate a predilection of the Lewis lung carcinoma for lymphatic pathways for metastasis throughout the animal body. The concomitant macrometastases to the visceral organs observed in this model may be remetastasis from the lymph nodes. This model of the Lewis lung carcinoma should be very useful in defining cellular trafficking and targeting mechanisms of metastasis, in particular those involving lymphatic pathways. 相似文献
9.
An imageable highly metastatic orthotopic red fluorescent protein model of pancreatic cancer 总被引:6,自引:0,他引:6
Katz MH Takimoto S Spivack D Moossa AR Hoffman RM Bouvet M 《Clinical & experimental metastasis》2004,21(1):7-12
In order to investigate the antitumor and antimetastatic efficacy of new chemotherapeutic agents, a novel, red-fluorescent, orthotopic model of pancreatic cancer was constructed in nude mice. MIA-PaCa-2 human pancreatic cancer cells were transduced with red fluorescent protein (RFP) and initially grown subcutaneously. Fluorescent tumor fragments were then transplanted onto the pancreas by surgical orthotopic implantation (SOI), facilitating high-resolution, real-time visualization of tumor and metastatic growth and dissemination in vivo. Tumor growth at the primary site was visible within the first postoperative week, while distant metastasis and the development of ascites became visible over the following week. This MIA-PaCa-2-RFP model produced extensive local disease and metastases to the retroperitoneum (100%), spleen (100%), intestinal and periportal lymph nodes (100%), liver (40%) and diaphragm (80%), and gave rise to malignant ascites and peritoneal carcinomatosis in 80% of cases. Growth and metastasis of tumor was more rapid and frequent than in previously described orthotopic pancreatic cancer models, leading to a median survival of only 21 days after tumor implantation. This unique, red fluorescent model rapidly and reliably simulates the highly aggressive course of human pancreatic cancer and can be easily non-invasively visualized in the live animal. The model can therefore be used for the discovery and evaluation of novel therapeutics for the treatment of this devastating disease. 相似文献
10.
Vladimir?Bobek Katarina?Kolostova Daniela?Pinterov Michael?Boubelik Ping?Jiang Meng?Yang Robert?M.?Hoffman
The Lewis lung tumor has been extensively studied in both syngeneic and allogeneic mouse models. However, its metastatic potential and mechanism are poorly understood. The aim of the present study was to develop a highly metastatic lymph-node targeting, imageable model of the Lewis lung carcinoma in a syngeneic host. We report here a syngeneic model of the Lewis lung carcinoma in which the carcinoma cells are labeled with green fluorescent protein (GFP). The tumor cells were transplanted in the dorsal side of the ear of C57-B16 mice in order to give the tumor cells access to the lymphatic system. This model of the Lewis lung carcinoma extensively metastasized to numerous lymph nodes throughout the body of the animal as well as visceral organs, as visualized by fluorescence microscopy using the bright GFP signal. Twenty-one different metastatic sites, including lymph nodes throughout the body, were identified among the cohort of transplanted animals. The data demonstrate a predilection of the Lewis lung carcinoma for lymphatic pathways for metastasis throughout the animal body. The concomitant macrometastases to the visceral organs observed in this model may be remetastasis from the lymph nodes. This model of the Lewis lung carcinoma should be very useful in defining cellular trafficking and targeting mechanisms of metastasis, in particular those involving lymphatic pathways.This revised version was published online in August 2005 with a corrected cover date. 相似文献
11.
A model of spontaneous lung metastases visualised in fresh host tissue by green fluorescent protein expression 总被引:3,自引:0,他引:3
Paris S Chauzy C Martin-Vandelet N Delpech B Thiberville L Martin JP Diarra-Mehrpour M 《Clinical & experimental metastasis》1999,17(10):817-822
The authors describe a model of spontaneous lung metastases in nude mice using green fluorescent protein (GFP) expression
as a marker. The human lung cell line H460M was transfected with the humanised GFP-S65T cDNA and a stable fluorescent cell
line termed H460MGFP was obtained. The latter kept in vitro biological features when compared to the parental H460M cell line, which suggests that GFP-expression does not influence
H460MGFP cell line behaviour. In order to evaluate their metastatic potential and to determine the number of spontaneous metastases,
H460MGFP cells were subcutaneously inoculated into nude mice. Animals were sacrificed at time intervals and tissues (lung, liver,
spleen, node, and kidney) were analysed under fluorescence microscopy. These experiments demonstrated that 2 weeks after subcutaneous
inoculation, 75% of animals exhibited fluorescent spontaneous lung micrometastases. From the third week, 100% of animals exhibited
an increasing number of metastases (10–16) which were only localised in the lungs. At the end of the study, the number of
lung metastases had dramatically increased (42–400 at 7 weeks). Although these metastases were mainly localised in lung, a
few mice had an invasion of neighbouring lymph nodes. The H460MGFP cell line allowed to follow the seeding and development of spontaneous lung metastases and may be considered a simple and
powerful tool to study each step of the metastasis to screen new anticancer drugs.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
12.
X-irradiation to the nude mouse lung and the ensuing pulmonary metastasis of the injected human cancer cells were investigated. Human cancer cells were injected intravenously into nude mice following 20 Gy right hemithoracic irradiation. The right lungs showed evidence of metastasis while metastasis was slight in the non-irradiated left lung. Platelet aggregation and fibrin deposition occurred around the arrested cancer cells in the capillaries of the right lung. The fibrinolytic activity of the irradiated right lung was lower than that of the non-irradiated contralateral lung. Natural killer cell activity was lower in the right-lung-irradiated mice than in the non-irradiated mice. We conclude that when the target organ is exposed to X-irradiation, there will be a decrease in fibrinolytic activity, a condition paving the way toward a metastasis. 相似文献
13.
Establishment of fluorescent lung carcinoma metastasis model and its real-time microscopic detection in SCID mice 总被引:5,自引:0,他引:5
Huang MS Wang TJ Liang CL Huang HM Yang IC Yi-Jan H Hsiao M 《Clinical & experimental metastasis》2002,19(4):359-369
Lung cancer is the most prevalent malignant tumor in the world. Metastasis of the disease causes death in lung cancer patients.
Recent study has shown that multiple cascades of gene defects occur in lung cancer. In this report, we established a novel
H1299/EGFP tumor model to determine whether H1299 transfected with the enhanced green fluorescent protein (EGFP) gene in vitro and xenotransplanted into SCID mouse lung would permit the detection of lung cancer micro-metastasis in vivo. We demonstrated that EGFP-transduced H1299 cells maintained stable high-level EGFP expressions during their growth in vivo. EGFP fluorescence clearly demarcated the primary seeding place and readily allowed for the visualization of distant micrometastasis
and local invasion at the single-cell level. Small metastatic and locally invasive foci, including those immediately adjacent
to the tumor's leading invasive edge, were almost undetectable by routine hematoxylin and eosin staining and immunohistochemistry.
The GFP tagged lung cancer model is superior for the detection and study of physiologically relevant patterns of lung cancer
invasion and metastasis in vivo.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
14.
An ultra-metastatic model of human colon cancer in nude mice 总被引:5,自引:0,他引:5
Fang X. Sun Aaron R. Sasson Ping Jiang Zili An Reza Gamagami Lingna Li A.R. Moossa Robert M. Hoffman 《Clinical & experimental metastasis》1999,17(1):51-58
An ultra-high metastatic model of human colon cancer was developed in order to represent highly malignant patient disease for which there is no current model. Surgical orthotopic implantation (SOI) of a histologically intact liver metastasis fragment derived from a surgical specimen of a patient with metastatic colon cancer was initially implanted in the colon, liver and subcutaneously in nude mice. This tumor did not metastasize for the first 10 passages. At the eleventh passage, the tumor exhibited metastasis from the colon to the liver, spleen, and lymph nodes. At this time, two selective passages were carried out by transplanting resulting liver metastases in the nude mice to the colon of additional nude mice. After these two passages, the tumor became stably ultra-metastatic and was termed AC3488UM. One-hundred percent of mice transplanted with AC3488UM with SOI to the colon exhibited local growth, regional invasion, and spontaneous metastasis to the liver, lymph nodes, and spleen. While the maximum size of the primary tumor was 0.9 g, the metastatic liver was over 9 times the weight of the normal liver with the maximum weight of the metastatic liver over 12 g. Liver metastases were detected by the tenth day after transplantation in all animals. Half the animals died of metastatic tumor 25 days after transplantation. Histological characteristics of AC3488UM tumor were poorly differentiated adenocarcinoma of colon. Mutant p53 is expressed heterogeneously in the primary tumor and more homogeneously in the liver metastasis suggesting a possible role of p53 in the liver metastasis. The human origin of AC3488UM was confirmed by positive fluorescence staining for in situ hybridization of human DNA. The AC3488 human colon-tumor model with its ultra-high metastatic capability in each transplanted animal, short latency and a short median survival period is different from any known human colon cancer model and will be an important tool for the study of and development of new therapy for highly metastatic human colon cancer. 相似文献
15.
Tiam1基因过表达对结直肠癌细胞增殖和转移能力的影响 总被引:1,自引:0,他引:1
目的探讨Tiaml(T lymphoma invasion and metastasis1)基因转染对结直肠癌细胞增殖、转移能力的影响。方法采用裸鼠皮下接种结直肠癌细胞的方法观察Tiaml基因对结直肠癌细胞体内增殖能力的影响,利用外科原位移植技术观察Tiaml对体内增殖、转移能力的影响。结果接种HT29/Tiaml细胞(转染Tiaml基因)的裸鼠皮下肿瘤生长明显加快,肿瘤体积自皮下注射后的第7天起与接种HT29/mock细胞(转染对照)的裸鼠皮下肿瘤比差异有统计学意义,接种HT29/Tiaml细胞的裸鼠皮下肿瘤第20天的平均体积是接种HT29/mock细胞组的2.5倍。进行结直肠癌细胞外科原位移植6周后,接种HT29/Tiaml细胞的裸鼠结肠原位肿瘤重量明显大于接种HT29/mock细胞的裸鼠(t=-14.916,P〈0.01)。采用外科原位移植的方法比较HT29/mock和HT29/Tiaml细胞的体内转移能力。在HT29/Tiaml细胞组,7只裸鼠全都发生了腹腔转移,4只发生了肝转移,其中有1只裸鼠发生了广泛转移,包括脾、肺及淋巴结等转移。在HT29/mock细胞组,7只中只有2只裸鼠发生了腹腔转移,没有一只裸鼠发生远处转移。结论Tiaml基因是结直肠癌增殖、转移的促进基因,Tiaml表达可作为结直肠癌增殖、转移过程中一个有价值的指标。 相似文献
16.
Yamamoto N Yang M Jiang P Xu M Tsuchiya H Tomita K Moossa AR Hoffman RM 《Clinical & experimental metastasis》2003,20(7):633-638
We have established stable, bright green fluorescent protein (GFP)- or red fluorescent protein (RFP)-expressing HT-1080 human
fibrosarcoma clones. These cell lines showed similar cell proliferation rates and high-frequency experimental lung metastasis.
The HT-1080-GFP and -RFP clones enable simultaneous real-time dual-color imaging in the live animal. HT-1080 cells were transduced
with retroviral vectors containing GFP or RFP and the neomycin resistance gene. Stable transformants were selected stepwise
with G418 up to 800 μl/ml. Subsequently, high GFP- or RFP-expressing clones, HT-1080-GFP or HT-1080-RFP, respectively, were
selected. 3×106 cells from each clone were mixed and injected into the tail vein of SCID mice. The cells seeded the lung at high frequency
with subsequent formation of pure green and pure red colonies as well as mixed yellow colonies with different patterns visualized
directly on excised lungs. The lung metastases were also visualized by external fluorescence imaging in live animals through
skin-flap windows over the chest wall. Lung metastases were observed on the lung surface of all mice. SCID mice well tolerated
multiple surgical procedures for direct-view imaging via skin-flap windows. Real-time metastatic growth of the two different
colored clones in the same lung was externally imaged with resolution and quantification of green, red, or yellow colonies
in live animals. The color coding enabled determination of whether the colonies grew clonally or were seeded as a mixture
with one cell type eventually dominating, or whether the colonies grew as a mixture. The simultaneous real-time dual-color
imaging of metastatic colonies described in this report gives rise to the possibility of color-coded imaging of clones of
cancer cells carrying various forms of gene of interest.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
17.
18.
Development of a green fluorescent protein metastatic-cancer chick-embryo drug-screen model 总被引:3,自引:0,他引:3
Bobek V Plachy J Pinterova D Kolostova K Boubelik M Jiang P Yang M Hoffman RM 《Clinical & experimental metastasis》2004,21(4):347-352
The chick-embryo model has been an important tool to study tumor growth, metastasis, and angiogenesis. However, an imageable model with a genetic fluorescent tag in the growing and spreading cancer cells that is stable over time has not been developed. We report here the development of such an imageable fluorescent chick-embryo metastatic cancer model with the use of green fluorescent protein (GFP). Lewis lung carcinoma cells, stably expressing GFP, were injected on the 12th day of incubation in the chick embryo. GFP-Lewis lung carcinoma metastases were visualized by fluorescence, after seven days additional incubation, in the brain, heart, and sternum of the developing chick embryo, with the most frequent site being the brain. The combination of streptokinase and gemcitabine was evaluated in this GFP metastatic model. Twelve-day-old chick embryos were injected intravenously with GFP-Lewis lung cancer cells, along with these two agents either alone or in combination. The streptokinase-gemcitabine combination inhibited metastases at all sites. The effective dose of gemcitabine was found to be 10 mg/kg and streptokinase 2000 IU per embryo. The data in this report suggest that this new stably fluorescent imageable metastatic-cancer chick-embryo model will enable rapid screening of new antimetastatic agents. 相似文献
19.
Henriksson KC Almgren MA Thurlow R Varki NM Chang CL 《Clinical & experimental metastasis》2004,21(6):563-570
Neuroblastoma is the most common extra-cranial solid tumor of infancy and childhood, and majority of patients die from the
metastatic disease. Orthotopic xenograft mouse models are valuable tools for improving our understanding and control of neuroblastoma
metastasis, because they readily represent genetic diversity and allow spontaneous metastasis. Intra-adrenal injection is
commonly used for establishing the orthotopic animal models since human neuroblastoma frequently originates in the adrenal
gland. However, it is unclear whether the metastatic potential of neuroblastoma can be reliably determined in adrenally-injected
mice because their gland size is so small. In this study, we developed and characterized a fluorescent orthotopic xenograft
animal model of NB69-derived human neuroblastoma. By comparing animals receiving adrenal injection and adrenal overlay, with
the latter mimicking injection spillover, we found that the metastatic potential of neuroblastoma can be reliably determined
in animal lungs. Furthermore, the lung metastasis can be genetically modulated in these animals. The results also show that
the expression of Renillagreen fluorescent protein (GFP) was exceptionally stable in NB69 cells, allowing rapid and sensitive detection of lung metastases
at the macroscopic level. Additional features of our model include 100 tumor take, a 1-week tumor latency, resemblance to
tumor behaviors in neuroblastoma patients, and the ability to monitor the expression of a gene of interest with GFP. This
animal model of human neuroblastoma will be useful for studying genes involved in the metastatic process and for evaluating
anti-metastasis agents in pre-clinical trials.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
20.
目的探讨血管性血友病因子(vWF)在不同转移潜能人肺癌裸鼠移植瘤中的表达。方法体外培养低转移及高转移人肺癌细胞系A549和D95,Transwell小室法检测肿瘤细胞体外侵袭和迁移能力;10只Balb/c裸小鼠随机分为A549组和D95组(n=5)。0. 2 mL两种肿瘤细胞悬液(浓度约为1×10^7/mL)分别皮下注入A549组及D95组裸小鼠左上肢,构建人肺癌裸鼠移植瘤模型。观察vWF对裸小鼠移植瘤生长的影响;ELISA检测外周血vWF水平;免疫组化检测移植瘤vWF蛋白表达;免疫印记检测移植瘤、肺和肝vWF蛋白表达。结果 D95肺癌细胞体外侵袭和迁移细胞数多于A549肺癌细胞(P<0. 05);D95组裸小鼠于皮下接种后第5天全部出现瘤结节,A549组于第7天全部出现;与A549组相比,D95组裸小鼠移植瘤质量及肺转移瘤结节数显著增多(P<0. 05),D95组移植瘤、肺及肝组织vWF蛋白的表达显著高于A549组(P<0. 01)。结论 vWF与人肺癌细胞的转移潜能密切相关。 相似文献