Bladder-cancer specific oncolytic adenovirus independent of CAR and establishment method

An oncolytic adenovirus and construction method technology, applied in the field of bladder cancer-specific oncolytic adenovirus and construction, can solve the problems of low expression and limited tumor cell efficiency

Inactive Publication Date: 2017-04-26
LANZHOU UNIVERSITY
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, the expression of CAR in poorly differentiated or highly malignant tumor cells is often low, which limits the efficiency of transfection of tumor cells with adenovirus type 5.

Method used

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  • Bladder-cancer specific oncolytic adenovirus independent of CAR and establishment method
  • Bladder-cancer specific oncolytic adenovirus independent of CAR and establishment method
  • Bladder-cancer specific oncolytic adenovirus independent of CAR and establishment method

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Embodiment Construction

[0021] The present invention will be further described in detail below with reference to the accompanying drawings, but it should not be construed as a limitation of the present invention. Without departing from the spirit and essence of the present invention, any modifications or substitutions made to the methods, steps or conditions of the present invention fall within the scope of the present invention. Unless otherwise specified, the technical means used in the embodiments are conventional means well known to those skilled in the art.

[0022] 1. Construction of Shuttle Plasmid

[0023] Synthetic DNA oligonucleotide fragments RO-301 and RO-302, the sequence of RO-301 is shown in SEQ ID NO.1, the sequence of RO-302 is shown in SEQ ID NO.2; RO-301 and RO- 302 were dissolved in water at 200mM each, mixed in equal volumes, heated and denatured at 95°C, and then slowly cooled to room temperature and refolded into a multi-cloning site linker; the MCS linker was inserted into th...

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Abstract

The invention discloses a bladder-cancer specific oncolytic adenovirus independent of CAR, an establishment method and application and belongs to the technical field of biological genetic engineering. Multiple cloning site linkers are made by artificially synthesizing DNA oligonucleotide fragments, expression of an oncolytic adenovirus EIA gene is controlled based on a specific promoter UPII of the urinary bladder, a prostate stem cell antigen enhancer PSCAE is inserted into the upstream of the promoter UPII to enhance the activity of the promoter UPII, and the bladder-cancer specific oncolytic adenovirus Ad5/F11p-PSCAE-UPII-E1A is established, can remarkably inhibit proliferation of CAR low-expression bladder cancer cells, has the bladder cancer specificity replication ability and has dose and time dependence.

Description

technical field [0001] The invention relates to the technical field of biogenetic engineering, in particular to a CAR-independent bladder cancer-specific oncolytic adenovirus, its construction method and application. Background technique [0002] Bladder cancer, one of the most common malignancies of the urinary system, has two distinct types: noninvasive (superficial) and invasive bladder cancer, both in terms of molecular mechanisms of formation and origin, and treatment It is completely different from other aspects such as recovery. At present, the clinical treatment of bladder tumors is mainly based on surgery, supplemented by radiotherapy, chemotherapy and bladder perfusion. However, the cure rate is far from satisfactory. Traditional chemotherapy and radiotherapy have disadvantages such as low anti-tumor ability, small killing index, and large side effects. Bladder cancer, especially muscle-invasive bladder cancer, is resistant to a variety of chemotherapy drugs, and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/861A61K39/235A61P35/00
CPCC12N7/00A61K39/00C12N15/86C12N2710/10021C12N2710/10032C12N2710/10043
Inventor 王志平曹文娟田俊强高彦俊郭琦
Owner LANZHOU UNIVERSITY
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