Three Kinds Of Mice Implanted Liver Tumor Model Building And Applied Research

Background:Primary hepatocellular carcinoma (Primary Hepatocellular Carcinoma, PHC) refers to occurred in human liver cells or intrahepatic bile duct cancer cell disease finest, is one of common malignant tumors in China. Our morbidity and mortality of liver cancer are the first in the world [1-2], after a very easy to relapse or metastasis [3-5], and resistance to chemotherapy,5-year survival rate is low[6-7] therefore requires in-depth study of the pathogenesis of liver cancer, to explore new methods of treatment. Many human studies due to the limitation of ethics, the need to establish an animal model in line with the development of features and variation of clinical disease and liver cancer. Mainly induced liver cancer in animal models of liver cancer and liver transplant two categories. Liver carcinogenesis model due to the long period of time, simply targeting a liver molding rate to be improved, both affect the application of liver carcinogenesis in animal models, so transplanted liver cancer model has become an important object of study.Currently, human liver cancer-liver cancer mouse model is the most widely used research animal model in which the common model of human liver cancer cell lines or tissue blocks inoculated in nude mice, severe combined immunodeficient mice (SCID) mice and other immune deficiencies, resolve Many problems of cancer recurrence, metastasis and treatment mechanism of basic research, but such model is characterized by immune deficiency, but the mice were used as liver cancer research "reactor", the lack of immune system factors for liver cancer. Therefore, this study focus on and build a different immune status and mice transplanted liver cancer model, a common approach to cancer recurrence and metastasis to the Walker-256 murine cell lines and human hepatoma cell lines HepG2 cell origin, by hepatic portal vein injection and injection, build a simulation liver metastasis, and mice implanted liver tumor recurrence model, this model try to simulate the behavior of the clinical features of malignant liver cancer, and clinical application of rat models in which radiofrequency ablation The experimental treatment and to determine the efficacy study. In addition, murine Walker-256 cell line is also secondary to metastatic liver cancer clinical simulation process occurs. In this study the metastasis of HCC recurrence and secondary metastatic liver cancer pathogenesis and intervention therapy in experimental animal models.Objective:Under normal immune status and immunosuppressive interventions to build three mice implanted liver tumor model, provide animal models for the study of metastatic and recurrent hepatocellular carcinoma, and try to rats used in clinical RFA treatment and determine the efficacy study.Methods:Select Mouse Walker-256 cell lines and human hepatoma cell line HepG2, respectively, for normal immune and liver transplantation in rats "triple immunosuppressive regimen" immunosuppressed mice, using direct injection and hepatic portal vein injection establish three kinds of rat orthotopic liver transplantation model, and the formation of the following groups:① portal-Walker-256 Group:Doors intravenous Walker-256-mice implanted liver tumor model, SD rats 30; ② lobe-Walker-256 Group:Walker-256-lobe injected mice implanted liver tumor model, SD rats 30; ③ portal-HepG2 group:the portal vein injection of human HepG2 hepatoma cells-immune suppression Balb/c mice were implanted liver tumor model, mouse 15, triple immunosuppressive regimen is methylprednisolone, tacrolimus and mycophenolate mofetil. Liver cancer mouse model outcome measures are:① animal general and rate of tumor, tumor size, liver metastasis and invasion of adjacent organs;③ Rats ultra high frequency ultrasound imaging observation; ② tumor tissue HE staining was observed:; ④ dynamic observation of mice weight, white blood cell count and classification. In addition, murine orthotopic liver transplantation model is also used for the experimental study of radiofrequency ablation, radiofrequency ablation of the lesion and take the surrounding tissue, conduct some detection:①:HE staining light microscope concept; ② apoptosis TUNEL staining.Results:Three methods can be successfully constructed murine orthotopic liver transplantation model, molding time is between 9 to 15 days, tumor formation rate between 14.3% to 33.3%, including high-frequency ultrasonic testing portal -Walker-256 group lobe -Walker-256 group rat model, can be found lobe tumors, mainly body is round or oval hypoechoic irregular groups, heterogeneous internal echo. HE staining under a microscope showed the tumor cells arranged in cords and lumpy, big stain nuclei, atypia, less cytoplasm, characteristic of metastatic liver cancer. Radiofrequency ablation of liver cancer model in rats after intervention to the needle point at the center region extending outward delivery times microscope obvious necrosis, apoptosis, necrosis and apoptosis based mixing zone area, of which the first two regions have a lot of inflammation cell invasion, the two regions have seen after a small amount of residual tumor cells. Portal vein-HepG2 liver cancer orthotopic transplantation model mice, mice with white blood cell count was 8.87±4.25%; the proportion of the value of neutrophils was 66.78 ± 14.49%, the ratio of the number of lymphocytes accounted 18.36±1.07%, Monocytes 9.45±.00%, the weight of mice showed a gradual downward trend, there were significant gaps normal. Pathological drawn tumor size (diameter) of 3.0mm. Pathological observation of light microscope, cell disorder, increased mitotic large nuclei, into a deep dyeing, prominent nucleoli, atypia, less cytoplasmic staining lighter and even colorless.Conclusions:Three implanted liver tumor model was successfully constructed, can basically simulate blood metastasis and metastasis of metastatic liver cancer and liver cancer recurrence, orthotopic liver transplantation model can be used to treat experimental studies of RF.