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Aim To establish subcutaneous and orthotopic transplantation models of human lung cancer in nude mice, and compare the anti-cancer effects of digoxin between the two models. Methods After subcutaneous inoculation of H460 tissues in nude mice, the tumor volume was measured; HE staining and immunohistochemistry were performed; H460-Luc cell suspension was injected into the lung of nude mice toestablish orthotopic tumor model, the in vivo imaging and fluorescence values were recorded, and the tumor lesions in other organs were observed after dissection. Results Compared with control group, the gemcitabine group had a significant anti-tumor effect (P 0.05). HE staining showed that the cell density in each treatment group decreased, and necrosis and/or fibrous hyperplasia were obvious. Immunohistochemistry indicated that the protein expression of p-p38, p-ERK and Nur77 in each treatment group significantly increased in the subcutaneous transplantation model; in the orthotopic transplantation model, the gemcitabine, the middle (P < 0.05) and low dose of digoxin group could inhibit the tumor growth, while the high dose of digoxin group accelerated the development of tumor (P < 0.05). Conclusion Digoxin is more sensitive to orthotopic transplanted tumor than subcutaneous transplanted tumor, anddigoxin may inhibit the tumor growth by up-regulating the expression of p-p38, pERK and Nur77.
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Objective To investigate the effect of SABP, a water-soluble component of Salvia miltiorrhiza, on the growth of orthotopic transplantation of H22 liver cancer and the immune microenvironment of liver cancer. Methods We established a mouse model of orthotopic transplantation of H22 cell liver cancer in BALB/c mice. ELISA was used to detect the expression of PD-L1, TGF-β, IL-1β, IL-10, IL-4, IFN-γ, IL-18, IL-7, IL-2, CCL-2 and CCL-21 in the liver. We counted the organ indexes of liver, spleen and kidney. Results SABP inhibited the growth of orthotopic transplantation tumors of H22 cell liver cancer, and increased the expression levels of PD-L1, TGF-β, IL-1β and IL-10 in the microenvironment of liver cancer, as well as the liver, spleen and kidney coefficients. Conclusion SABP could inhibit the growth of orthotopic transplantation tumors of H22 cell liver cancer and promote the expression of PD-L1, TGF-β, IL-1β and IL-10 in the microenvironment of liver cancer.
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Objective To investigate the effect of medical sodium hyaluronate gel (HA) on the growth and metastasis of abdominal and pelvic tumor cells in vitro and in nude mice. Methods Three tumor cells, Hela, CT26 and HCT116, were used to investigate the effects of different HA concentrations on the growth and migration of tumor cells in vitro by MTT assay and Transwell assay. An orthotopic transplantation model of colonic tumor in nude mice was established to investigate the effect on the proliferation of cell HCT116 by comparing the tumor volume and tumor mass 4 weeks after inoculation. The effects on the metastasis of cell CT26 were investigated by comparing the tumor metastasis rate and the number of metastatic lesions of lung and liver in nude mice among the different experimental groups 3 weeks after inoculation. Results HA did not promote the growth and metastasis of Hela, CT26 and HCT116 cells in vitro at different concentrations. Actually, HA exhibited a certain inhibitory activity at the concentration of 5 mg/ml. In the orthotopic transplantation model of colonic tumor-HCT116, HA did not promote the growth of cell HCT116. In the orthotopic transplantation model of colonic tumor-CT26, HA inhibited CT26 tumor metastasis. Conclusion Under the experimental conditions, HA did not promote the growth, migration or metastasis of abdominal and pelvic related tumor cells including Hela, CT26 and HCT116 in vitro and in vivo.
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Background and purpose: Renal cell carcinoma is the most common form of kidney cancer, characterized by lack of early symptoms and high malignancy. This study aimed to establish orthotopic nude mice models of human renal cell carcinoma with high success rate and good repeatability. Methods: The four types of methods which were adopted to establish the orthotopic models of renal cell carcinoma were orthotopic injection of 786-0 and ACHN cell suspensions, orthotopic injection of primary cell suspensions obtained from the subcutaneous tumor tissues, renal subcutis orthotopic implantation into renal capsule and surgical subcutis orthotopic implantation into renal fascia. To gain insights into the tumorigenicity and the growth of transplantation tumors, the imageological examination (PET/CT), histological examination (H-E staining, immunohistochemistry staining) and biochemical analysis of blood were carried out. Results: In terms of the subcutaneous transplantation of human renal cell carcinoma models in nude mice, tumorigenic rate of ACHN cells (90%) was higher than that of 786-0 cells (30%). The tumorigenic incidences of 786-0 cell suspensions orthotopic injection, ACHN cell suspensions orthotopic injection, ACHN subcutis cellular suspensions orthotopic injection, ACHN subcutis orthotopic implantation into renal capsule and renal fascia were 33%, 80%, 90%, 100% and 20%, respectively. ACHN subcutis orthotopic implantation into renal capsule was the most effective approach. Imageological and histological results accorded with poorly differentiated renal cell carcinoma. Conclusion: Four orthotopic nude mice models of human renal cell carcinoma were successfully established. Among these methods, ACHN subcutis orthotopic implantation into renal capsule is the most effective approach, which provides an ideal model for the research on biological behavior of human renal cell carcinoma and its treatment.
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Objective To study the feasibility of DCE-MRI application in the orthotopic transplantation model of gastric cancer in nude mice.Methods Orthotopic transplantation model of gastric cancer in nude mice was established, and 15 models underwent DCE-MRI using the contrast agent of gadopentetate.Subsequently, the tumor was dissected in order to detect the MVD.The MVD was compared between orthotopic transplantation tumor model of gastric cancer and normal gastric mucosa.Results Fifteen nude mice with orthotopic transplantation of gastric cancer successfully underwent DCE-MRI examination.As for the cancer, the values of Ktrans, Kep and Ve were (2.11±0.44) min-1, (4.59±0.93) min-1, and 0.46±0.06, respectively.The MVD in gastric cancer tissues was significantly higher than that in normal gastric mucosa (χ2=16.205, P<0.001).Conclusion DCE-MRI can be used for noninvasively quantitative evaluation of vascular parameters of gastric carcinoma.
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@#The study aims to establish a human gastric cancer orthotopic transplantation model in nude mice and to use 7T MRI for detection. After poorly differentiated human gastric adenocarcinoma SGC-7901 cells were injected subcutaneosly into the right flanks of nude mice, the model of in nude mice was established with orthotopic transplanted cancer of gastric tumor by the Compont® gel pasted method. 7T MRI scan was conducted on the mice after operating model about 20 days later. Histopathological examinations were carried out on the stomach. Two of three mice on which 7T MRI scan were performed showed visible suspected stomach tumor and their presence was verified again by histopathological examinations; tumor formation rate in the nude mice gastric orthotopic transplantation model was 66. 7%. This study suggested that 7T MRI could be used in the live detection of in situ tumor and that MRI could be used for pre-clinical gastric cancer drug development and clinical gastric carcinoma diagnosis.
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Objective:To explore the influence of different cell culture methods in the genome-wide DNA methylation status of breast cancer MDA-MB-231 cells,and to clarify the relationship between genome-wide DNA methylation status and cell growth environment and the role of genome-wide DNA methylation status in the occurrence and development of tumor.Methods:The MDA-MB-231 cells were cultured with 2D and 3D cell culture models and mouse orthotopic transplantation model (Ti model)and collected, then DNA was extracted by DNA extraction kit and the genome-wide DNA methylation status of MDA-MB-231 cells after cultured with three different culture methods was detected by DNA methylation chip,then the value of beta,DiffScore and Delta_Beta of the CpG loci of each gene were calculated by applying Genomestudio software, and the differential methylation genes were screened by Genomestudio software and GO and Pathway analysis of these genes were performed in DAVID on-line analysis tool.Results:All 480 genes of the MDA-MB-231 cells showed significant differences in the degree of methylation in 3D and 2D models (P<0.05);86 448 genes in 3D and Ti models (P<0.05);90 005 genes in Ti and 2D models (P<0.05).The differential methylation genes in 3D and 2D,3D and Ti,and Ti and 2D models were enriched on the multicellular organismal development term and cell differentiation term (P<0.05);also on MAPK signaling pathway,cell adhesion molecules (CAMs),and regulation of actin cytoskeleton (P<0.05). Conclusion:There are differences in genome-wide DNA methylation status of MDA-MB-231 cells cultured in 2D, 3D cell culture and Ti models.
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Objective To investigate the characters and targeted ability of FITC-CSNRDARRC molecular probe in labeling orthotopic transplantation tumor of bladder cancer in vivo.Methods From July 2013 to June 2014,the stability and characters of FITC-CSNRDARRC molecular probe were detected by spectrophotometer and molecular imaging and the optimum concentration and imaging time window were determined.30 BALB-C nude mice were randomly divided into experimental group (n =20) and control group(n =10).In control group,5 of them (group A) were ligated bilateral ureter,others(group B) were not.We established orthotropic transplanted bladder tumor (BIU-87) model by operation.And 0.2 ml probes (220 μmol/L) was then injected intravenously in all mice after 2 weeks.We obtained images and analyzed average gray value of the heart,lung,liver,spleen,bilateral kidney and orthotropic transplantation bladder tumor by using optical probe molecule fluorescence imaging system after 30 min,1 h,2 h,4 h and 12 h,respectively.Results After injected the FITC-CSNRDARRC molecular probes intravenously at 220 μmol/L,the fluorescence signal of tumor tissue strengthened gradually.The optimal imaging time window was 4 hours after injection.The illumination and temperature had little effect on the fluorescence signal.With the time passing after injection,the intensity of florescence signal progressively increasing,which reached the peak at4 h.The average gray value of tumor tissue at 1 h,2 h,3 h,4 h,5 h,6 h,8 h,12 h were 74.22,76.2,80.11,89.38,83.29,85.1,81.22,83.01,respectively.The fluorescence signal of normal tissue weakened gradually with the passage of time.Only liver and gall bladder could notice the fluorescence signal 4 hours after injection in group A.However,the relatively strong fluorescence signal could be found in liver and gall bladder in group B.Conclusions The characters of fluorescence probe are affected by its concentration.Its optimal concentration of labeling tumor is 220 μmol/L.The optical imaging time window was about 4 h after intravenous injection.The FITC-CSNRDARRC molecular probe can specifically bound to orthotopic transplanted tumor of bladder cancer in vivo.
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Objective By comparing the biological characteristics among Lewis lung cancer cells ( LLC) , LLC or-thotopic transplantation derivative cells ( R1-LLC) and R1-LLC orthotopic transplantation derivative cells ( R2-LLC) , we e-valuate their invasion and metastatic abilities in orthotopic transplantation models.Methods In vitro, we applied CCK8, clonogenic assay, and Transwell invasion assay to detect the proliferation ability, invasion ability and morphologic structures of LLC,R1-LLC, R2-LLC cells respectively, meanwhile observing morphologic structures by transmission electron microsco-py.In vivo, we constructed LLC, R1-LLC and R2-LLC orthotopic transplantation models.Herein, we observed their tumor growth and metastasis.The tumor formation rate and tumor-forming time were recorded for statistic analysis.Results In vitro:LLC, R1-LLC and R2-LLC cells showed no significant difference in proliferation ability ( P>0.05 ) , but significant differ-ences in invasion ability:R2-LLC>R1-LLC>LLC(P<0.05).In vivo:In the orthotopic group, the tumor formation rates of LLC, R1-LLC and R2-LLC cells were 66.67%、80%、93.33%(P <0.05).Conclusions In orthotopic implantation mouse models, R2-LLC cells present higher invasion and metastatic ability than R2-LLC and LLC cells.
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Objective To establish an orthotopic osteosarcoma nude mice model that co-expresses green fluorescent protein( GFP) , red fluorescent protein ( RFP) and luciferase for the purpose of monitoring the growth of osteosarcoma and screening drug candidates against osteosarcoma .Methods Human osteosarcoma cells of U 2-OS were infected with lentivirus carrying reporter gene .The reporter gene expression was verified by fluorescent microscopy and bioluminescence imaging.The cells were transplanted into tibia of the nude mice and monitored by bioluminescence imaging .Results The reporter gene was stably expressed in U 2-OS cells.The growth and metastsis of osteosarcoma could be detected in nude mice.Conclusion The established orthotopic osteosarcoma nude mice model is an ideal model for investigating the mechanism of growth and metastasis of osteosarcoma and for screening drug candidates against osteosarcoma .
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Objective To establish a stable orthotopic model with high green fluorescent protein (GFP) expression in nude mice and observe its biological features.Methods Human HCT116 colon cancer cells transfected with GFP pLPCX retroviral plasmid were used to build a subcutaneous tumor model in nude mice.Fifteen BALB/C nude mice were selected to underwent orthotopic transplantation of colon when the GFP-labeled tumor grew to 10 mm × 10 mm as observed by in vivo fluorescent microscopy.The growth and metastasis of orthotopically implanted colon cancer cells were observed with fluorescent imaging system at different time points.The differences of the tumor size measured by peripheral vernier caliper and fluorescent imaging system were analyzed using the t test,and the differences in different groups were analyzed using the analysis of variance.Results GFP-labeled colon cancer models were successfully established in all the 15 nude mice,and there was no surgery-related complications or death.Tumors marked by GFP were observed under fluoroscope in week 3.The size of the tumors progressively increased with time.The volumes of the orthotopically transplanted tumors obtained from global measurement using fluorescent imaging system were greater than those measured by peripheral vernier calipers at postoperative week 3,4,5,6,7,while no statistically significant difference was observed (t =-1.280,-1.115,-0.718,-0.199,-0.386,P ＞0.05).There was a significant difference in the interation of measure method and different time points (F =29.546,P ＜ 0.05).Eight nude mice survived at the end of the experiment,and tumor metastasis was observed in 6 mice.Conclusions It is technically feasible to construct GFP-labeled colon cancer orthotopic transplantation model.The mice model could be used for real-time,in vivo,non-invasive and dynamic observation and analysis of the growth and metastasis of tumor cells.
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Objective To establish a stable model of rat orthotopic left lung transplantation using direct suture of vessels and bronchi. Methods Ten Sprague Dawley (SD) rats weighted 250 to 350 g were used as lung donors and recipients respectively. Airway and pulmonary vessels were reconstructed microsurgically using continuous running suture technique. Survival time were recorded and donor lungs were checked by autopsy. Results All 10 rats received left lung transplantation were weaned from ventilator successfully. Both of cold ischemia time and warm ischemia time were about 40 minutes. The total procedure took about 130 minutes. Autopsy was used to check the patency of anastomotic sites. No thrombosis or air leak was found. Conclusions Direct microsurgical surture can be used to establish an experimental model of orthotopic left allograft lung transplantation in rats. This method is proved to be stable, reliable and similar to clinical practices.
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Objective:To establish the pulmonary metastasis model of osteosarcoma in rats.Methods:UMR106 cells(rat osteosarcoma strain) were cultured in RPMI1640 media and collected before transplantation and resuspended in serum-free media or collagen gel to a final concentration of 1?107 cells/ml.About 200?l cell suspensions were injected into the right proximal tibia of 3-week-old Sprague-Dawley rats.Twelve rats were used in every group sorted by resuspension fluid.All rats were treated by cyclosporin A and SMZ-TMP after transplantation.The dates of tumor appearance and rat death were noted,the size of tumor was measured once a week,the metastasis foci on the lung surface was counted and the limbs afflicted by tumor were observed by X-ray after death.Both the tumors in limb and lung were stained with hematoxylin and eosin,and analyzed by microscopy.Results:All rats developed local intratibial bone tumors that were radiographically and histologically similar to primary human osteosarcoma.The pulmonary metastasis foci were found in all rats at autopsy.The tumor formation time of serum-free media group was(10.0?1.65) days,shorter than that of collagen gel group,which was(17.8?0.87) days(P
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Objective To evaluate the inhibitory effect of rofecoxib in human gastric carcinoma tissues established in nude mice by orthotopic transplantation through the expressions of vascular endothelial growth factor (VEGF) and inducible nitric oxide synthase (iNOS). Methods After the models of human gastric carcinoma in nude mice by orthotopic transplantation were established, rofecoxib was administered intragastrically. The expressions of VEGF and iNOS were evaluated in the local tumors by Envision immunohistochemical method. Results The expression of VEGF protein in isotonic saline group, rofecoxib group and combination group were 81.25%, 61.25%, and 35.00% respectively.There was significant difference between rofecoxib group and isotonic saline group, and so was rofecoxib group and combination group (P
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PURPOSE: The metastatic animal model of human cancer is important from a practical point of view in the research of cancer metastasis, because it resembles the original tumors morphologically, biologically and biochemically. We developed the animal model to investigate the clinically relevant metastasis of gastric cancer which is the leading cause of death in Korea. METHODS: Seven to eight-week-old specific-pathogen-free (SPF) BALB/c-nu mice were used. We developed an orthotopic transplantation model using the tissue obtained from an inoculation of the gastric cancer cell suspension (YCC-3) into a subcutaneous layer of mice. The mice were kept in laminar-flow cabinets under SPF condition and inspected everyday. RESULTS: Mice were sacrificed 8~12 weeks after the operation when they showed either a measurable mass or signs of distress. The metastatic pattern of this animal model was very similar to that of human gastric cancer. At autopsy, the local growth of the gastric cancer, lymph node metastasis and any distant metastasis were noted. CONCLUSION: We developed an animal model for human gastric cancer metastasis that will enhance our understanding of the biology of cancer metastasis and it will contribute to the development of the research and treatment of cancer metastasis.
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Animais , Humanos , Camundongos , Autopsia , Biologia , Causas de Morte , Coreia (Geográfico) , Linfonodos , Modelos Animais , Metástase Neoplásica , Neoplasias GástricasRESUMO
Objective To investigate the inhibitory effect of earthworm fibrinolytic enzyme (EFE) on the metastasis of human hepatoma cells.Methods A metastatic model of human hepatocellular carcinoma in nude mice via orthotopic implantation was established.After the modeling for 7 days,the mice were divided randomly into the control group,low-dose EFE group (800 uku/kg?d-1),and high-dose EFE group (1600 uku/kg?d-1).After administration for 30 days,the implanted tumors were weighted,and the intrahepatic transmission rate and abdominal cavity seeding rate were calculated after examination by naked eyes.Then the pulmonary metastasis were examined under microscope,and the expression of focus adhesion kinase (FAK) and ?1-intigrin were detected through RT-PCR and western blotting method.Results Compared with the model control group,the mean weight of the orthotopic tumor was reduced,and the intrahepatic transmission rate and abdominal cavity seeding rate were decreased in FEF groups (P