Gene drug for combined treatment on tumors, and use thereof

A tumor and composition technology, applied in anti-tumor drugs, gene therapy, drug combinations, etc., can solve the problems of ineffective, difficult and weak tumor treatment response, achieve broad application prospects, good biological safety, and promote tumor cells. the effect of apoptosis

Inactive Publication Date: 2013-02-13
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in most cases, the body's response to tumor treatment is ineffective and weak
The main reason is that the occurrence and development of malignant tumors is a long-term, multi-stage, multi-gene change accumulation proces...

Method used

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  • Gene drug for combined treatment on tumors, and use thereof
  • Gene drug for combined treatment on tumors, and use thereof
  • Gene drug for combined treatment on tumors, and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Embodiment 1, the preparation of recombinant adenovirus

[0062] 1. Amplification of Endostatin gene and TRAIL gene

[0063] Specific primers were designed according to human Endostatin gene and human TRAIL gene,

[0064] The primer sequence of amplification Endostatin gene is as follows:

[0065] Upstream primers:

[0066] 5'-GCTCTAGAGCCACCATGTACAGGATGCAACTCCTGTC-3'(XbaI),

[0067] Downstream primers:

[0068] 5'-GGGGTACCTTACTTGGAGGCAGTCATG-3'(KpnI),

[0069] The primer sequences for amplifying the TRAIL gene are as follows:

[0070] Upstream primers:

[0071] 5'-GCTCTAGAGCCACCATGTACAGGATGCAACTCCTGTC-3'(Xba I),

[0072] Downstream primers:

[0073] 5'-GGGGTACCTTTAGCCAACTAAAAAGGCCC-3'(Kpn I),

[0074] Using the T vector carrying the Endostatin gene (sequence 1 in the sequence listing can also be used as a template) and the TRAIL gene (sequence 2 in the sequence listing can also be used as a template) as a template, the above primers are used to perform PCR ampl...

Embodiment 2

[0096] Embodiment 2, Ad-Endostatin and Ad-TRAIL recombinant adenovirus combined effect

[0097] 1. Detection of the inhibition efficiency of endothelial cells and tumor cells by single action

[0098] HUVEC and HepG2 cells were cultured in 96-well plates, and when the cell fusion rate reached 80%, they were infected with 10 MOI and 50 MOI of recombinant adenovirus Ad-Endostatin and Ad-TRAIL respectively according to the method of Example 1. After 24 hours, 48 ​​hours and 72 hours after transfection, add 10 μl of 5 mg / ml MTT solution to each well, continue to incubate for 4 hours, remove the culture medium, add 100 μl DMSO to dissolve for 10 minutes, and detect the solution on a microplate reader The absorbance at 490nm was used to calculate the proliferative activity of the cells. Take an equal volume of 0.1M PBS, pH 7.4 PBS and 10MOI or 50MOI equivalent Ad-EGFP as a control.

[0099] The maximum value in each experimental PBS treatment group is 100%; value-added rate (%)=(A...

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Abstract

The invention discloses a gene drug for combined treatment on tumors, and a use thereof. The gene drug is a composition for preventing and/or treating tumors. The composition is composed of 1), Endostatin protein coding gene-containing recombinant viruses and TRAIL protein coding gene-containing recombinant viruses, or 2), an Endostatin protein and a TRAIL protein, or 3), Endostatin protein coding gene-containing recombinant vectors and TRAIL protein coding gene-containing recombinant vectors. An experiment proves that in the gene drug provided by the invention, an Endostatin gene for inhibiting tumor neovascularization and a TRAIL gene for promoting tumor cell apoptosis respectively carry signal peptides and are used for construction of adenovirus high-expression vectors. The gene drug can be used for treating a cervical cancer, a liver cancer, a breast cancer and a lung cancer.

Description

technical field [0001] The invention belongs to the field of gene therapy, and in particular relates to a gene drug for combined treatment of tumors and its application. Background technique [0002] The rapid proliferation and malignant metastasis of tumor benefit from the increase of tumor angiogenesis and the decrease of tumor cell apoptosis and other factors. The key genes or target proteins in the corresponding pathways of various factors are being studied by scientists more and more and successfully applied in clinic. However, in most cases, the body's response to cancer treatment is ineffective and weak. The main reason is that the occurrence and development of malignant tumors is a long-term, multi-stage, multi-gene change accumulation process. The tumor is affected by multiple signaling pathways in the process of confrontation and selection with the body. A single change of a certain gene or It is often difficult to achieve satisfactory results for a certain signa...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K39/00A61K38/17A61P35/00C12N15/63C12N7/01
Inventor 黄来强严飞
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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