Application of recombinant adenovirus in producing antineoplastic medicine

A recombinant adenovirus and anti-tumor drug technology, applied in the field of biology, can solve the problems of difficult to achieve effective concentration in vitro, unable to exert drug effect, insufficient drug intake, etc.

Inactive Publication Date: 2008-02-27
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The multidrug resistance characteristics of tumors can be summarized into four types: generally including one, insufficient effective drug concentration in cells: (1) insufficient drug intake; (2) drug pumping: chemotherapy absorbed by cells If the drug is re-pumped before it takes effect, the drug effect cannot be exerted, which is another reason for the insufficient concentration of the effective drug in the cell
(3) Intracellular drugs are "blocked" so that they c

Method used

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  • Application of recombinant adenovirus in producing antineoplastic medicine
  • Application of recombinant adenovirus in producing antineoplastic medicine
  • Application of recombinant adenovirus in producing antineoplastic medicine

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Experimental program
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Embodiment 1

[0035] Embodiment 1: Construction and preparation of recombinant adenovirus (Ad-WWOX)

[0036] (1) Construction of recombinant adenovirus

[0037] Replication-deficient adenovirus AdMax adenovirus packaging system (product of MicrobixBiosystems, Canada, the map of the shuttle plasmid—pDC316 plasmid is shown in Figure 1, and the backbone plasmid—pBHGlox_E1, 3Cre.)

[0038] 293 cells are products of MICROBIX BIOSYSTEMS, Canada.

[0039] 1. Construction of pDC316-IRES-EGFP plasmid

[0040] 1.1 Amplified sequence IRES-EGFP from plasmid pIRES-EGFP

[0041] Primers were designed (primer design software: oligo6.0), the sequence IRES-EGFP was amplified from the plasmid pIRES-EGFP (purchased from clontech company), and the plasmid pDC316-IRES-EGFP was constructed.

[0042] The primer sequences are:

[0043] IRES up 5'-GCCAGTCGACACCGCATCGAGCTGA-3'

[0044] IRES down 5'-CTGCCAAGTTGCTCGAAGTCGACT-3'

[0045] Using primers IRES up and IRES down to amplify fragment IRES-EGFP from pIRES...

Embodiment 2

[0156] Example 2: Experimental Study of Recombinant Adenovirus Mediated Human WWOX Gene on Drug Resistance Gene Therapy of Human Lung Adenocarcinoma Cells

[0157] (1) Detection of drug resistance gene therapy in lung adenocarcinoma drug-resistant cells (in vitro drug sensitivity test-CCK-8 method)

[0158]A549 / T+ group, A549 / T-group and A549-group grew up to 80% infected with Ad (MOI: 50), collected cells in 48 hours; added to 96-well culture plate (100ul complete medium+8000 cells / well), Incubate for 12 hours, add different concentrations of CDDP (Australian Keding Pharmaceutical Co., Ltd.) or TAX (three replicate wells for each well); incubate in a 37°C incubator for 72 hours, suck up the disposable infusion needle connected to the vacuum pump to measure Well culture solution, add complete medium (premixed 5ul CCK-8+100ul medium containing 10% fetal bovine / well), cultivate for 3.5 hours; measure the absorbance at 450nm (reference wavelength 650nm), calculate the cell surviv...

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Abstract

The invention discloses an application of recombinant adenovirus of human-carrying WWOX gene to prepare antineoplastic drug in the molecular biological domain, which is characterized by the following: constructing recombinant adenovirus of human-carrying WWOX gene; establishing drug tolerant model A549 Paclitaxel drug-tolerant strain and gene transmission method; observing the expression of adenovirus in the A549/T cell; reversing the expression of Paclitaxel and cisplatin through WWOX in the external drug sensitive test. The lung cancer is malignant tumour with highest mortality based on general chemotherapy as main therapeutic method, but most of NSCLC displays insensitivity corresponding to chemotherapy in vivo and in vitro. The invention can reverse the drug tolerance of lung cancer cell, which provides new method to solve the drug tolerant problem of malignant tumour.

Description

technical field [0001] The invention relates to the field of biology, in particular to the application of a recombinant adenovirus carrying human WWOX gene in the preparation of antitumor drugs, in particular to the preparation of chemotherapeutic drug sensitizers. Background technique [0002] Lung cancer is a malignant tumor with a high mortality rate. The main reasons are high incidence, low possibility of surgical cure and poor response to chemotherapy. 80% of lung cancer patients are non-small cell lung cancer, and about 40% of non-small cell lung cancer is diagnosed at an advanced stage, systemic chemotherapy has become the main treatment. But we have to face the fact that the vast majority of non-small cell lung cancers (NSCLC) have been shown to be chemotherapy insensitive both in vivo and in vitro. [0003] One of the biggest obstacles in tumor chemotherapy is the resistance of tumor cells to drugs. Some tumors, such as colorectal cancer cells, have inherent resis...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61P35/00A61P11/00C12N15/861
Inventor 李强吴俊杰
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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