Application of annexin A3 in preparation of drug for target killing liver cancer stem cells

A liver cancer stem cell and annexin technology is applied in the application field of ANXA3 in the preparation of targeted killing liver cancer stem cell drugs, which can solve the problem of lack of clear antigen and achieve the effect of broad clinical application prospects.

Inactive Publication Date: 2015-03-25
夏建川
View PDF0 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, previous studies have not clarified which antigen plays a major role
There are no research reports on liver cancer
At present, there is no research report on HCC stem cell-associated antigens

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of annexin A3 in preparation of drug for target killing liver cancer stem cells
  • Application of annexin A3 in preparation of drug for target killing liver cancer stem cells
  • Application of annexin A3 in preparation of drug for target killing liver cancer stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Isolation and identification of liver cancer stem cell-associated antigens

[0021] Because CD133 has been considered as a marker of liver cancer stem cells, in this example, a gene chip was used to screen the gene profiles differentially expressed in CD133+ and CD133- liver cancer cells. CD133+ liver cancer stem cells and CD133- non-liver cancer stem cells were sorted by flow cytometry, and their total RNA was extracted, and the high-expression CD133+ liver cancer stem cells were preliminarily screened by Affymetrix Human Gene 1.0ST human gene expression profile chip scanning analysis. Gene, real-time fluorescence quantitative polymerase chain reaction and immunoblotting methods further verified that the screened ANXA3 gene was highly expressed in CD133+ liver cancer stem cells.

[0022] (1) Materials and methods:

[0023] 1) cell line

[0024] Huh7 cells were purchased from the Japan Health Science Research Resource Bank, SMMC-7721 cells were purchased fr...

Embodiment 2

[0041] Example 2: The role of ANXA3 in maintaining the biological characteristics of liver cancer stem cells

[0042] (1) Materials and methods:

[0043] 1) Establish a liver cancer cell line that stably overexpresses and silences ANXA3

[0044] The recombinant lentivirus expressing ANXA3 and 5 μg / mL polybrene were added to the well-growing liver cancer cells. After 48 hours, the liver cancer cell lines stably overexpressing ANXA3 were screened with fresh medium containing 2 μg / mL puromycin. The ANXA3-interfering plasmid was transfected into liver cancer cells with liposome 2000. After 48 hours, the liver cancer cell lines with stable ANXA3 silence were screened with fresh medium containing 2 μg / mL puromycin.

[0045] 2) Ball formation experiment

[0046] Single cells (5,000 cells / ml) were plated in a six-well ultra-low adsorption culture plate, treated with 20ng / mL human recombinant epithelial growth factor, 10ng / mL human recombinant basic fibroblast growth factor, 2% B27 a...

Embodiment 3

[0057] Example 3: DC cells loaded with ANXA3 antigen

[0058] (1) Materials and methods:

[0059] In view of the important role of ANXA3 in maintaining the biological characteristics of liver cancer stem cells, we used it as an antigen associated with liver cancer stem cells, loaded DC cells, and prepared ANXA3 antigen-specific CTL cells.

[0060] Buffy coat cells (from Guangzhou blood station) of HLA-A11 positive healthy people were collected, and mononuclear cells (PBMC) were obtained by density gradient centrifugation. Use Quanta-007 serum-free medium to suspend and isolate PBMC, spread on 75cm 2 culture flask in CO 2 Incubate in the incubator for 1 hour, shake the cell culture flask gently, collect the non-adherent suspension cells and maintain the culture with Quanta-007 serum-free medium containing 20 U / mL IL-2. Adhered cells were added to DC cell culture medium (CellGro DC culture medium + 1000U / ml GM-CSF + 400U / ml IL-4) for DC cell culture. On the 5th day, DC cells ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to novel use of annexin A3 (ANXA3) and particularly relates to an application of annexin A3 in preparation of a drug for target killing liver cancer stem cells. The invention puts forward the application of using ANXA3 as a liver cancer stem cell related antigen and then inducing to obtain the drug for target killing liver cancer stem cells by using liver cancer stem cell antigen-loaded ANXA3 for the first time. Anti-hepatoma specific immunotherapy based on the drug has a wide clinical application prospect.

Description

technical field [0001] The invention belongs to the fields of molecular biology, cell biology, tumor immunology and tumor immunotherapy, and relates to a new application of annexin A3 (ANXA3), in particular to the application of ANXA3 in the preparation of drugs targeted to kill liver cancer stem cells. Background technique [0002] It is currently believed that tumor recurrence and metastasis are related to a group of heterogeneous cells called "tumor stem cells", which have the ability to self-renew, differentiate and induce tumor formation. Many research results have shown that cancer stem cells are not sensitive to conventional radiotherapy and chemotherapy, and are prone to tolerance. Therefore, traditional radiotherapy and chemotherapy mainly kill ordinary tumor cells, but have no killing effect on tumor stem cells, and these tumor stem cells are thus enriched. , and then gain a proliferative advantage, eventually triggering tumor recurrence and metastasis. [0003] m...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): A61K39/00A61K35/26A61P35/00
Inventor 夏建川潘求忠
Owner 夏建川
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap