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Use of hepatitis B virus x protein and encoding gene thereof in preparing medicament for treating cancer of liver

A technology for hepatitis B virus and liver cancer treatment, applied in gene therapy, drug combination, genetic engineering and other directions, can solve the problem of no hepatitis B virus HBx protein, etc., and achieve good results and good therapeutic effects

Active Publication Date: 2009-08-12
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is currently no report in the field that hepatitis B virus HBx protein is used to prevent and treat chronic hepatitis, liver cancer and its complications

Method used

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  • Use of hepatitis B virus x protein and encoding gene thereof in preparing medicament for treating cancer of liver
  • Use of hepatitis B virus x protein and encoding gene thereof in preparing medicament for treating cancer of liver
  • Use of hepatitis B virus x protein and encoding gene thereof in preparing medicament for treating cancer of liver

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Preparation of recombinant adenovirus Ad-X

[0052] For a brief technical route of the construction process, see figure 1 . It mainly includes the following steps

[0053] a. Use Gateway technology to connect the hepatitis B virus X protein gene into the gate vector using Nco1 and Xho1 restriction sites;

[0054] b. Using the principle of gene recombination, the entry vector containing the X protein gene and the vector containing the adenovirus genome are subjected to in vitro recombination under the action of recombinase (please verify the statement of recombination) to construct a gene containing the target and the recombinant plasmid of the adenovirus genome (E1, E3 deletion);

[0055] c. Transfect the recombinant plasmid containing the target gene and the adenovirus genome (E1, E3 deletion) into 293 cells after linearization with Pac I, and package it into a recombinant adenovirus (Ad-X) containing the target gene;

[0056] d. Packaging the recombinant...

Embodiment 2

[0184] Example 2 Efficacy experiment of X protein recombinant adenovirus of the present invention

[0185] The main equipment used:

[0186] 1) PCR instrument: Hybaid Company, model: PxE.

[0187] 2) Constant temperature shaker: Forma Company, model: 4580.

[0188] 3) Gel imaging system: Bio-Rad Company, model: Fluor-S.

[0189] 4) Electroporator: Bio-Rad Company, model: Gene Pulser Xcell.

[0190] 5) Biological safety cabinet: Forma Company, model: A / B3.

[0191] 6) CO2 incubator: Forma Company, model: 3111.

[0192] 7) Liquid nitrogen tank: Forma Company, model: 8031.

[0193] 8) Inverted phase contrast microscope: Olympus Company, model: CK40.

[0194] 9) Inverted phase contrast microscope: Nikon Company, model: TE2000U.

[0195] 10) Refrigerated high-speed centrifuge: Beckman Company, model: J25.

[0196] 11) Refrigerated ultracentrifuge: Beckman Company, model: Optima L-100XP.

[0197] 12) Ultra-low temperature refrigerator: SANYO Company, model: MDF-382E.

[0198...

Embodiment 3

[0230] Example 3 Experimental research on the therapeutic effect of X protein recombinant adenovirus against liver cancer ascites of the present invention

[0231] 1. Method

[0232] 1.1 Culture of mouse liver cancer cell line (H22) Take mouse liver cancer H22 cells stored in liquid nitrogen, revive at 37°C, and use 1640 medium containing 10% fetal bovine serum at 37°C, 5% CO 2 cultivated in an incubator. After the cells are full, collect the cells, wash the cells with serum-free 1640 medium, count the cells, and take 1×10 7 H22 cells were inoculated into the peritoneal cavity of mice and passaged three times in the peritoneal cavity.

[0233] 1.2 Establishment of the ascites model Take an appropriate amount of ascites, count the cells, and dilute the ascites with normal saline to a concentration of 2.5×10 6 / ml, each mouse was intraperitoneally inoculated with 5×10 cells 5 / indivual.

[0234] 1.3 Treatment of malignant ascites Three days after inoculation, ascites formed...

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Abstract

The invention belongs to the field of gene therapy of diseases, relating to application of hepatitis B virus x protein gene (HBx) in preparing medicines for treating liver cancer and complicating diseases coupled with the liver cancer. The invention has the technical proposal to provide the application of the hepatitis B virus x protein gene (HBx) and encoded protein thereof in preparing the medicines for treating liver cancer and complicating diseases coupled with the liver cancer. In the invention, the hepatitis B virus x protein gene is recombined into an adenoviral vector to be used in medicines for treating the liver cancer; furthermore, activating transcription factors which are generally considered as virus promoters in the field are creatively engaged in virus pathogenic HBx protein for preparing medicines for treating the liver cancer and are proved to have good effect by in vitro and in vivo experiments, thus being well capable of treating the liver cancer and liver ascites,a complicating disease thereof and providing a new selection for liver cancer treatment in the field.

Description

technical field [0001] The invention belongs to the field of disease gene therapy. It relates to the use of hepatitis B virus X protein gene (HBx) in the preparation of medicines for treating liver cancer and liver cancer complications. Background technique [0002] Tumor is a common disease that seriously endangers human health and people's lives. Liver cancer is one of the most common malignant tumors. The time from the discovery of liver cancer to death is generally less than half a year, and it has become one of the main causes of death for human beings. China is a country with a large incidence of liver cancer, accounting for 45% of the world's morbidity and mortality. The incidence rate of liver cancer in my country ranks second among cancers, and it is the second cancer killer. Therefore, it is of great significance to study drugs for the treatment of liver cancer. With the rapid development of molecular biology, immunology, and cell biology, tumor biotherapy repre...

Claims

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Application Information

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IPC IPC(8): A61K38/16A61K48/00A61P1/16A61P35/00C12N15/51C12N15/861
Inventor 魏于全李玉华文艳君赵霞杨莉
Owner SICHUAN UNIV
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