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Novel lung specificity transfer liver cancer cell and preparation thereof

A liver cancer cell and specific technology, applied in the fields of biology and oncology, can solve the problems of large differences in the expression of specific markers, poor tumorigenicity, and difficulty in culturing tumor cells

Active Publication Date: 2018-08-31
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Tumor cell lines play an important role in the basic research of tumors. It is difficult to culture tumor cells in vitro, especially to establish human tumor cell lines that can grow and pass down for a long time, and have certain characteristics. Poor sex, inconsistent genetic background, or large differences in the expression of specific markers

Method used

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  • Novel lung specificity transfer liver cancer cell and preparation thereof
  • Novel lung specificity transfer liver cancer cell and preparation thereof
  • Novel lung specificity transfer liver cancer cell and preparation thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Example 1 Screening and identification of lung-specific metastatic liver cancer cell lines

[0079] The cells were first marked with GFP for the isolation of lung metastatic cells. Secondly, the cells were modified to stably express luciferase enzyme, which can be used for mouse live imaging to detect cell location and quantitative analysis. At the same time, the modified cells were labeled while constructing luciferase enzyme m-cherry, as a selection marker for luciferase-positive cells. Compared with the original hep-3B cells, the migration ability of the cells was significantly enhanced.

[0080] Specifically, luciferase-labeled hep-3B cells were injected into nude mice through the tail vein, and randomly metastatic liver cancer lesions appeared in the mice in 20-40 days, that is, the primary hep-3B cells did not have the organ specificity of metastasis. Among the randomly transferred mice, the mice with target organ metastasis were selected, the tumor mass of the l...

Embodiment 2

[0082] Example 2 Establishment and identification of a mouse model of lung-specific metastasis of liver cancer cells

[0083] 2.1 will be 5×10 5 -1×10 6 A lung-specific metastatic liver cancer cell (or its progeny cells) was inoculated in nude mice for 21-42 days, the lungs of the mice were taken out, shredded, digested with collagen enzyme to obtain a single-cell suspension, and cultured in DMEM medium until the cells adhered to the wall. GFP-positive cells were sorted by flow cytometry to obtain lung-specific metastatic liver cancer cells.

[0084] 2.2 Passage activity identification

[0085] It has been identified that the subculture activity of the cells is good, and the specific lung metastasis can still be stable after 15-20 passages, such as figure 2 b.

Embodiment 3

[0086] Example 3 Gene expression profile identification of lung-specific metastatic liver cancer cells

[0087]After culturing hep-3B cells and lung metastasis-specific cells for 2 passages, some of the cells were sent for preservation, and some of the cells in the parallel passages were identified for genetic characteristics, including gene sequencing, differential detection of miRNA profiles and mRNA profiles, cell adhesion, and When the culture dish reaches 50-70%, trypsinize, collect cells by centrifugation, lyse the cells by trizol method, extract RNA, reverse transcribe and perform mRNA expression chip (Shanghai Bohao Biotechnology Co., Ltd.) detection and gene sequencing. As a result, it was found that the genetic characteristics of the preserved cells and each progeny (2-15 generations) were the same.

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Abstract

The invention relates to a lung specificity transfer liver cancer cell 3B-LM. Particularly, the preservation number of the cell 3B-LM in China Center for Type Culture Collection is CCTCC NO:C2016173,and the cell belongs to an HEP-3B organ specificity transfer cell line LM.

Description

technical field [0001] The invention belongs to the fields of biology and oncology, in particular, the invention relates to a lung-specific metastatic liver cancer cell. Background technique [0002] Worldwide, hepatocellular carcinoma is the second leading cause of cancer-related death. The high metastases of liver cancer lead to poor prognosis for patients with liver cancer after drug treatment. The common clinical liver cancer metastasis is intrahepatic metastasis, lung metastasis and lung metastasis. transfer. [0003] The metastasis of liver cancer involves multiple processes, including shedding of tumor cells in situ in the liver, invasion of surrounding tissues, entry into the blood system, survival in the blood system, and formation of clones in distant tissues and organs. Each process involves tumor The changes in the cell itself and the interaction between the tumor and the microenvironment, the specific mechanism of action is not completely clear. [0004] Tumor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12Q1/02A01K67/027A61K49/00C12R1/91
CPCA01K67/0271A01K2207/12A01K2267/0331A61K49/0008C12N5/0693C12Q1/6886C12Q2600/158G01N33/5011
Inventor 王慧李晓光高志虎
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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