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Dual-targeting gene treatment vector for liver cancer and preparation method thereof

A dual-targeting, gene therapy technology, applied in the field of gene carriers, can solve the problems of low carrier specificity and large cell toxicity

Inactive Publication Date: 2011-09-28
SHANXI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the high positive charge density makes PEI exhibit great cytotoxicity at the same time, and the specificity of the carrier is not strong

Method used

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  • Dual-targeting gene treatment vector for liver cancer and preparation method thereof
  • Dual-targeting gene treatment vector for liver cancer and preparation method thereof
  • Dual-targeting gene treatment vector for liver cancer and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: Preparation of dual targeting vectors.

[0020] 1. Prepare the pAFP-TK-IRES2-EGFP plasmid vector at a concentration of 50 μg / mL.

[0021] 2. Place the chylomicrons in a sonicator and sonicate 10 times, 15s each time, with a power of 150w.

[0022] 3. According to the ratio of plasmid vector to chylomicrons (mass / volume) of 2 μg / mL, mix the products of step 1 and step 2, vortex for 30 seconds, and let stand for 30 minutes to prepare a dual-targeting carrier.

Embodiment 2

[0023] Example 2: Encapsulation effect of chylomicrons on plasmid vectors.

[0024] The dual-targeting carrier obtained in Example 1 was observed by scanning electron microscopy, and the results showed that when the ratio (mass / volume) of the plasmid carrier to chylomicron was 2 μg / mL, the encapsulation effect of the chylomicron on the plasmid carrier was the best. And the particle size is between 200-300nm, such as figure 2 .

Embodiment 3

[0025] Example 3: Protective effect of chylomicrons on plasmid vectors.

[0026] Add bovine pancreatic deoxyribonuclease I to the dual-targeting carrier obtained in Example 1 to make a final concentration of 20 μg / mL, digest at 37°C for 30 minutes, use a simple plasmid carrier as a control, and observe chyle by agarose gel electrophoresis Protection of plasmid vectors by microparticles. Observation see image 3 , Lane 1 shows that the pure plasmid vector is degraded by bovine pancreatic deoxyribonuclease Ⅰ because there is no protective effect of chylomicrons; lane 2 shows that chylomicrons in the dual-targeting vector have a strong protective effect on the plasmid vector, which can avoid its Degraded by bovine pancreatic deoxyribonuclease I. The results showed that the chylomicrons in the dual-targeting vector had a strong protective effect on the plasmid vector and could prevent it from being degraded, so the dual-targeting vector had high stability.

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Abstract

The invention relates to a dual-targeting gene treatment vector for liver cancer, which is a complex consisting of chylomicron and an eukaryotic expression plasmid vector containing an alpha-fetoprotein (AFP) promoter and a suicide gene. Apolipoprotein E (apo E) is contained in the chylomicron, and apo E receptors present on membrane surfaces of liver cells and liver cancer cells, and can identity and combine the apo E specifically, so the plasmid vector can be carried by the chylomicron specifically to be oriented to the liver cells and the liver cancer cells without being absorbed by other cells of bodies; and the eukaryotic expression plasmid vector contains the AFP promoter for promoting gene expression, and the promoter can be promoted only under the specific AFP promoting environment to promote the gene expression at the downstream of the eukaryotic expression plasmid vector, so the dual-targeting gene treatment vector has high and obvious liver cell targeting, and can be promoted and expressed only in the liver cancer cells.

Description

technical field [0001] The invention relates to a gene carrier, in particular to a gene carrier for carrying a target gene for treating liver cancer, and a preparation method of the gene carrier. Background technique [0002] Liver cancer is one of the most common tumors, and its incidence is increasing worldwide, seriously endangering human life and health. The current methods of treating liver cancer mainly include chemotherapy, surgery, intervention and other means, but the effect is not satisfactory. With the development of genetic science and technology, gene therapy for liver cancer has become possible. The key issue of gene therapy is the carrier of the gene. At present, the technology in this field is developing rapidly. The commonly used gene therapy carriers for liver cancer mainly include viral vectors, liposomes, plasmids, and cationic polymers. [0003] Viral vectors have high efficiency, but poor specificity, and due to safety issues, the application has not ...

Claims

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Application Information

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IPC IPC(8): A61K47/42A61P35/00A61K48/00
Inventor 解军于保锋徐钧司海东张小曼程凯王惠珍张悦红赵虹刘志贞张俊涛弓韬胡晓年向前
Owner SHANXI MEDICAL UNIV
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