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Establishment and application of hepatocellular carcinoma cell line HCC-LY5

A technology of HCC-LY5 and liver cancer cells, applied in the field of its establishment, a new hepatocellular carcinoma cell line HCC-LY5, which can solve the problems of non-expression, inconsistent cell line characteristics, and unstable passage

Inactive Publication Date: 2015-06-24
SHANGHAI INST OF ONCOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] For the field of liver cancer, at present, although there are some liver cancer cell lines, these liver cancer cell lines have many disadvantages, including: unstable passage, poor tumorigenicity, inconsistent characteristics of cell lines, expression of liver cancer-specific markers Jagged, some even non-expressive, etc., and therefore unsatisfactory

Method used

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  • Establishment and application of hepatocellular carcinoma cell line HCC-LY5
  • Establishment and application of hepatocellular carcinoma cell line HCC-LY5
  • Establishment and application of hepatocellular carcinoma cell line HCC-LY5

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Establishment of hepatocellular carcinoma cell line HCC-LY5

[0075] (a) Source

[0076] This cell line comes from a male liver cancer patient (patient number: 603320), and its clinical information is shown in the table below.

[0077] Table 1

[0078]

[0079] (b) Method

[0080] Take the primary liver cancer tissue of the liver cancer patient and inoculate it in immunodeficient mice, and take the tumor-forming tissue for repeated inoculation and passage, and finally obtain a stable passage cell line. It includes the following processes:

[0081]

[0082] In addition, in the sixth month, 5 male NOD / SCID mice were surgically incised for the third-generation mice. In the 7th month, bilateral lymph nodes were observed to be enlarged after tumor resection. After dissection, the lungs were transparent. The primary culture was performed. The lymph nodes were taken to pass 4 NOD / SCID mice, and the lungs were taken to pass 4 NOD / SCID mice. mouse. On the 8th month, the tumor was cut a...

Embodiment 2

[0088] Biological characteristics of human liver cancer cell line HCC-LY5

[0089] In this example, the biological characteristics of the purebred human hepatocarcinoma cell line with the deposit number of CCTCC NO: C201099 were detected by conventional methods. The results are as follows:

[0090] 2.1 Cell morphology:

[0091] Under the inverted phase contrast microscope, the HCC-LY5 liver cancer cells are polygonal or circular, uniform in shape, and passaged every 72 hours ( image 3 ).

[0092] 2.2 Enzyme / green fluorescent protein double labeling

[0093] The result of luciferase / GFP double labeling of HCC-LY5 cell line is as follows Figure 4 (Right panel) shows that all cells are labeled with green fluorescent protein (GFP).

[0094] 2.3 HCC-LY5 cell immunocytochemical analysis

[0095] HCC-LY5 liver cancer cells were planted on a cell chip, and the expression of liver cell-related proteins was analyzed by immunocytochemistry. The antibodies used and the dilution are shown in Table...

Embodiment 3

[0131] Establishing an animal model of liver cancer with human liver cancer cell lines

[0132] Take the human hepatocellular carcinoma cell line HCC-LY5 (preservation number: CCTCC C201099) obtained in Example 1, after digestion with the digestive solution (0.25% pancreatin, 0.02% EDTA, pH 7.3), collect and count, and count it as 1×10 6 Or 1×10 7 The number of cells / mouse was subcutaneously inoculated into 20 4-6 weeks old Balb / c nude mice on the right chest wall of the mouse.

[0133] Between 14-60 days after vaccination, all 20 mice developed tumors one after another. The pathological examination was performed in the same way as in Example 2, and it was confirmed that hepatocellular carcinoma was induced in the animal model.

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Abstract

The invention provides establishment and application of a hepatocellular carcinoma cell line HCC-LY5. Specifically, the hepatocellular carcinoma cell line HCC-LY5 provided in the invention has stable traits, can be subcultured stably and repeatedly, and is applicable to established animal models of hepatocellular carcinoma, thus being an ideal cell line for the basic and preclinical phase application study of hepatocellular carcinoma. In addition, a very small part of the hepatocellular carcinoma cell line HCC-LY5 expresses hepatocellular carcinoma stem cell marker CD133. The invention also provides application of the cell line in establishing model animals, screening drugs, and other aspects.

Description

Technical field [0001] The present invention relates to the fields of biology and oncology, and more specifically to a new hepatocellular carcinoma cell line HCC-LY5, and its establishment method and application. Background technique [0002] The occurrence and development of various human diseases are very complicated. It is necessary to thoroughly explore the pathogenesis and curative effects of various diseases and cannot be tested on patients. However, it is possible to follow the Animal Protection Law and conduct research on various diseases and life phenomena through animals, and then deduce that humans explore the mysteries of human life, explore the laws of human disease occurrence and development, so as to control the occurrence and development of human diseases, and cure diseases and diseases. Alleviate the suffering of human diseases, improve the quality of life, and prolong human life. [0003] For a long time, people have found that it is difficult to promote the deve...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09A61D7/00A01K67/027A61K49/00
Inventor 李锦军姚明闫明霞张立行孙荷芬刘蕾葛超郝向芳顾健人
Owner SHANGHAI INST OF ONCOLOGY
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