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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 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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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 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 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