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Establishment and application of mmhrl1 transgenic mouse liver tumor cell line

A liver tumor and transgenic technology, applied in the fields of biology and oncology, can solve the problems of difficulty in establishing mouse liver tumor cell lines, high experimental costs, and complex influencing factors.

Inactive Publication Date: 2020-05-08
DALIAN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the high cost, time-consuming and labor-intensive experiments in mice, and the complex influencing factors, it is urgent to establish corresponding mouse liver tumor cell lines
However, in contrast, the establishment of mouse liver tumor cell lines is more difficult

Method used

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  • Establishment and application of mmhrl1 transgenic mouse liver tumor cell line
  • Establishment and application of mmhrl1 transgenic mouse liver tumor cell line
  • Establishment and application of mmhrl1 transgenic mouse liver tumor cell line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Establishment of liver tumor cell MMHRL1 in H-ras12V transgenic male mice

[0059] (a) source

[0060] The H-ras12V transgenic male mouse liver tumor cell line comes from H-ras12V transgenic male mice, and its specific information is shown in the table below.

[0061] Table 1

[0062]

[0063] The primary liver tumor tissue of the H-ras12V transgenic male mouse was obtained, and the liver tumor tissue was directly cultured in vitro, digested and passaged repeatedly, and finally a stable passage H-ras12V transgenic male mouse liver tumor cell line was obtained. Specifically include the following processes:

[0064] The above-mentioned H-ras12V transgenic male mice were sacrificed painlessly, and the liver tumors were excised by aseptic technique. Place the liver tumor in a 10cm culture dish filled with sterilized normal saline at normal temperature, fully clean the tumor capsule, and use a scalpel to cut the liver tumor tissue collected after the above operation to...

Embodiment 2

[0068] Biological characteristics of liver tumor cell MMHRL1 in H-ras12V transgenic male mice

[0069] In this example, conventional methods were used to detect the biological characteristics of the purebred H-ras12V transgenic male mouse liver tumor cell MMHRL1 with the preservation number CCTCC NO: C2016103, and the results are as follows:

[0070] 2.1 Cell Morphology

[0071] Observing H-ras12V transgenic male mouse liver tumor cells MMHRL1 under an ordinary optical microscope, it can be seen that most of the liver tumor cells are round or polygonal, and the cell shape is uniform ( figure 2 A). Cell Counting Kit-8 (CCK8 kit) was used to detect the proliferation state of the cell line, and the operation was carried out in full accordance with the operation guide of the kit ( figure 2 B). The cells proliferated stably.

[0072] 2. Genotype identification of 2H-ras12V transgene

[0073] Take the H-ras12V transgenic male mouse liver tumor cell MMHRL1 (preservation number...

Embodiment 3

[0086] Establishment of liver tumor animal model using H-ras12V transgenic male mouse liver tumor cell MMHRL1

[0087] The H-ras12V transgenic male mouse liver tumor cell MMHRL1 (preservation number: C2016103) obtained in Example 1 was collected and counted after being digested with digestive solution (0.25% trypsin, 0.02% EDTA, pH7.3), and counted according to 1×10 6 or 1×10 8 The number of cells / mouse was inoculated into the subcutaneous site of the right armpit of the mice, and the inoculated mice were 5 nude mice at the age of 5 weeks.

[0088] Between 14 and 60 days after inoculation, tumors occurred successively in five mice. The same method as in Example 2 was used for biological detection, and it was confirmed that hepatocellular liver tumors were induced in animal models. The result is as Figure 5 shown.

[0089] Cell line deposit

[0090] The H-ras12V transgenic male mouse liver tumor cell MMHRL1 of the present invention was deposited in the China Center for T...

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Abstract

The invention provides establishment and application of an H-ras 12V transgenic male mouse liver tumor cell line MMHRL1. Specifically, the H-ras 12V transgenic male mouse liver tumor cell line MMHRL1 provided by the invention has stable properties, can be stably passed for a plurality of times, is applicable to establishment of a liver tumor animal model and is an ideal cell line for foundation and clinical prophase application researches of liver tumor. Furthermore, the H-ras 12V transgenic male mouse liver tumor cell line MMHRL1 can be used for expressing H-ras mutated cancer genes and has highly-activated Ras / MAPK (Renin-angiotensin system / Mitogen-activated Protein Kinase) and PI3K / AKT / mTOR (Phosphoinositide 3-kinase / protein kinase B / mammalian Target of Rapamycin) signal paths. The invention further provides the application of the cell line to the aspects of establishment of model animals, drug screening and the like.

Description

technical field [0001] The present invention relates to the fields of biology and oncology, and more specifically relates to a novel H-ras12V transgenic male mouse liver tumor cell line MMHRL1, its establishment method and application. Background technique [0002] The occurrence and development of human diseases is a very complex process. In-depth study of the pathogenesis of various diseases and the effect of clinical drug treatment in order to achieve the purpose of preventing and curing various diseases is the unremitting goal of human beings. However, based on humanitarianism, direct experiments on patients cannot be used. In accordance with the Animal Protection Law, animal experiments can be used to conduct in-depth research on the occurrence and development of various diseases and life phenomena, so as to achieve the purpose of controlling the occurrence and development of human diseases, relieving patients' pain, and finally curing diseases. [0003] In the long-t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10A01K67/027A61K49/00
CPCA01K67/0271A01K2207/12A01K2227/105A01K2267/0331A61K49/0008C12N5/0693C12N2510/00
Inventor 王爱国刘要甫樊婷婷王靖宇王福金
Owner DALIAN MEDICAL UNIVERSITY
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