Orthotopic transplantation rat liver cancer model and preparing method and application thereof

Abstract

The invention pertains to the field of biomedical technology, in particular to an orthotopic transplanted mouse liver cancer model which takes the research of the occurrence and development mechanisms of liver cancer and the development of anti-tumor drugs as the purposes, and the preparation method and the application thereof. The H22 cell strain of stable transfection EGFP is injected into the BALB / c mouse peritoneal cavity for amplification; the ascites is extracted from the H22 ascites cancer mouse peritoneal cavity, then cell separation is carried out and cell suspension is prepared; the mouse peritoneal cavity is opened, and the cell suspension is extracted by a micro-syringe to be injected into the mouse liver; a cotton bud with 75 percent alcohol is pressed on the needle hole till the surface of the liver does not bleed any more, and the needle hole is sealed by bonding agent; then the liver surface is washed by physiological saline, and layer separation is carried out and the peritoneal cavity is closed. The results show that the occurrence rate of mouse liver cancer is 100 percent and the natural remission rate is zero. The model is an ideal orthotopic liver cancer model which can be used in the research of the development and metastasis mechanisms of orthotopic liver cancer and can also be applicable to the development of anti-tumor drugs, the experimental treatment and diagnosis research of liver cancer.

Description

1. Technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to a mouse orthotopic transplanted liver cancer model for the purpose of studying the molecular mechanism of liver cancer development and the development of antitumor drugs, as well as its preparation method and application. 2. Background technology [0002] Primary liver cancer is one of the common malignant tumors in my country, and its mortality rate is high, ranking third in the death order of malignant tumors. About 110,000 people die from liver cancer every year in my country, accounting for 45% of the death toll from liver cancer in the world. The treatment of liver cancer needs to be improved urgently. An ideal animal model of liver cancer is an indispensable experimental tool for the development of anti-tumor drugs, the mechanism of liver cancer development, diagnosis and treatment. Therefore, the successful establishment of experimental animal models of ...

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Problems solved by technology

In the experiment of Han et al., the production procedure is cumbersome and the success rate is low, and the model needs to be subcutaneously implanted in mice to form solid tumors, so the tumor tissue blocks transplanted in each experiment cannot come from the same mouse body, and the cut tissue The size of the block is not easy to be consistent, the content is not uniform, and it contains other non-cancerous tissues, which will seriously affect the repeatability and reliability of the experiment, and cannot fully reflect the effect of anti-liver cancer drugs

Li Qi et al. and Shao Chengwei et al. established a rat transplanted liver cancer model by intrahepatic injection of Walker-256 cells. The Walker-256 cell line used in the model can better simulate the expansion and infiltration of human liver cancer after implantation into the liver. However, the cell line is derived from the spontaneous carcinosarcoma of Wistar rat mammary glands rather than liver cancer tissue. Therefore, this model cannot well simulate the occurrence and development of liver cancer, let alone meet the requirements of anti-liver cancer drug development and screening, liver cancer molecular biology research needs

In addition, rats are more expensive and require a larger breeding space, so it is not easy to produce in large quantities

Although the existing human-nude mouse xenograft liver cancer model has a high transplantation success rate, the immune system of nude mice is deficient, which cannot reflect the real situation of normal organisms, and it is impossible to study the role of the immune system in the occurrence of liver cancer and the The impact on the subsequent development of liver cancer cannot simulate the normal metabolism of drugs in the body

In addition, the nude mouse-human liver cancer transplantation model requires high technical conditions, the postoperative survival rate of nude mice is low, and the requirements for sterility are high, feeding is difficult, and the price is expensive. Therefore, the practicability of this model is greatly limited.

At the same time, liver cancer models made by simply transplanting tumor tissue blocks or cell suspensions into the liver are not enough to serve as a platform for studying the molecular mechanism of liver cancer development and finding molecular targets for anti-liver cancer drugs.

Therefore, the existing animal models of liver cancer have certain defects and cannot meet the needs of the current biomedical field.

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