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Application of Clusterin in preparation of cancer therapy medicine for killing tumor stem cell

A tumor stem cell and cancer treatment technology, which is applied in the application field of cancer treatment drugs, can solve the problems of Clusterin that have not been reported in the literature and the mechanism of action is unknown, so as to achieve good application prospects, improve the effect of curing tumors, and improving the status quo of treatment

Active Publication Date: 2018-02-23
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the mechanism of action of Clusterin in tumors is currently unknown, and the expression and function of Clusterin in tumor stem cells are unknown to those skilled in the art. So far, Clusterin has not been used as a therapeutic target for tumor stem cells or in the preparation of drugs for treating tumor stem cells. Literature report

Method used

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  • Application of Clusterin in preparation of cancer therapy medicine for killing tumor stem cell
  • Application of Clusterin in preparation of cancer therapy medicine for killing tumor stem cell
  • Application of Clusterin in preparation of cancer therapy medicine for killing tumor stem cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1 Analysis of the expression of Clusterin during the growth and sphere formation of gastric cancer stem cells

[0060] 1. Identification and quantitative analysis of GCSC spheroidizing medium supernatant protein by iTRAQ multiple chemical labeling tandem mass spectrometry

[0061]Collect the GCSC culture medium (respectively denoted as D1-D5) and uncultivated medium (denoted as D0) cultured for 1-5 days, centrifuge to get the supernatant, concentrate the protein by ultrafiltration, trypsin (Trypsin) enzymolysis, enzymolysis The peptides were labeled and mixed with iTRAQ reagent (Applied Biosystems) and then analyzed by liquid phase tandem mass spectrometry (LC / MS / MS). Protein identification and quantitative analysis were performed by the ProteinPilot software platform (Applied Biosystems).

[0062] 2. Western blot

[0063] The protein concentration in the supernatant was determined by the BCA method, the same amount of protein samples were subjected to 10% SDS-...

Embodiment 2

[0068] Example 2 Clusterin is a key molecule regulating the stemness and survival of gastric cancer stem cells

[0069] 1. Establishment and identification of GCSC cells stably expressing inducible overexpression of Clusterin and HSP90, and GCSC cells stably expressing inducible Clusterin shRNA

[0070] Restriction endonuclease EcoRI, MluI double enzyme digestion PCR amplification Clusterin, HSP90 (from GCSC cells) gene products were purified and recovered, respectively connected to the inducible eukaryotic expression vector pLVX-TetOne-Puro digested by the corresponding endonuclease (Clontech Company), construct the recombinant expression vector pLVX-TetOne-Puro-Clusterin, pLVX-TetOne-Puro-HSP90. The empty control vector is pLVX-TetOne-Puro.

[0071] Clusterin and HSP90 overexpression viruses are composed of pLVX-TetOne-Puro-Clusterin, pLVX-TetOne-Puro-HSP90 and control pLVX-TetOne-Puro overexpression plasmids, respectively, plus packaging plasmid Lenti-X PackagingSingle Sho...

Embodiment 3

[0090] Example 3 Clusterin knockdown can significantly inhibit the growth of gastric cancer stem cell xenografts in nude mice

[0091] Female Balb / C nude mice (6 weeks old, body weight 18.0±2.0g) were cultured in a sterile environment. Mice were divided into two groups, Dox treatment group (Dox+) and control group (Dox-), 5 mice in each group, respectively injected 100 μl containing 4×10 5 The suspension of shClu2 GCSC cells (referred to as shClu) and shCtrl cells (referred to as shCtrl) was used to establish a subcutaneous xenograft tumor model. After 14 days, the tumor volume of the nude mice bearing the tumor reached 0.6cm 3After about 10 minutes, related treatments were started, wherein the Dox treatment group (Dox+) added Dox (1mg / ml, 0.5% glucose) to the drinking water; the control group (Dox-) did not add Dox (0.5% glucose) to the drinking water, and changed every 3 days. Drink water once, a total of 12 days. Closely observe the growth of the subcutaneous tumor, meas...

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Abstract

The invention discloses application of Clusterin in preparation of a cancer therapy medicine for killing a tumor stem cell. The invention also discloses a Clusterin inhibitor and the cancer therapy medicine. The invention further discloses a cancer combination therapy medicine. The cancer combination therapy medicine comprises Clusterin proteins and HSP90 proteins, and the cancer combination therapy medicine can kill the tumor stem cell. The invention discloses, for the first time, Clusterin which can be taken for preparing a molecular targeted therapy medicine for the tumor stem cell and canbe combined with HSP90 to prepare a molecular therapy medicine for the tumor stem cell, thereby contributing to development of a targeted medicine for the tumor stem cell, improving the current statusof malignant tumor therapy and curing tumors, and having a potential and great application prospect in the field of tumor therapy.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of Clusterin in the preparation of cancer treatment drugs for killing tumor stem cells. Background technique [0002] Malignant tumors are diseases with high morbidity and high mortality that seriously endanger human health. For example, gastric cancer (GC) is the most common malignant tumor of the digestive system, its incidence rate ranks fifth among all malignant tumors worldwide, its mortality rate ranks third among malignant tumors, and it also has a high recurrence rate and low treatment rates. [0003] Tumor cells in tumors can be divided into two types: one is normal tumor cells and the other is tumor stem cells. Ordinary tumor cells have the characteristics of rapid division, sensitivity to anticancer drugs, and no self-renewal ability. Cancer stem cells (Cancer stem cells, CSCs) are a small number of cell subsets in tumors that have the characteristics of...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61K45/00A61K45/06A61P35/00A61P35/04A61K38/16A61K31/409
CPCA61K31/409A61K38/16A61K38/1703A61K45/00A61K45/06A61K2300/00
Inventor 熊继先
Owner SHENZHEN UNIV
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