Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

SOD2 lentiviral vector, construction method and application thereof in targeted radiation protection and tumor sensitization

A technology of lentiviral vector and construction method, which is applied in the application field of SOD2 lentiviral vector, targeted radiation protection and tumor sensitization, can solve the problems of restricting the popularization and application of SOD2 gene therapy, so as to increase sensitivity and reduce apoptosis Mortality Effect

Inactive Publication Date: 2017-11-21
SICHUAN CANCER HOSPITAL
View PDF2 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the lack of effective gene transfection methods targeting the radiation field and the regulation of the expression of the introduced genes, the clinical application of SOD2 gene therapy is limited.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • SOD2 lentiviral vector, construction method and application thereof in targeted radiation protection and tumor sensitization
  • SOD2 lentiviral vector, construction method and application thereof in targeted radiation protection and tumor sensitization
  • SOD2 lentiviral vector, construction method and application thereof in targeted radiation protection and tumor sensitization

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] SOD2 lentiviral vector pLVX-C 9 The construction method of BC-SOD2-T2A-AcGFP1 comprises the following steps:

[0047] (1) Synthesis of CArG 9 , CMV basic promoter chimeric promoter C 9 BC gene fragment, the C 9 The upstream of the BC gene fragment contains a Cla I restriction site, and the downstream contains an Xho I restriction site;

[0048] (2), Cla I, Xho I double enzyme digestion lentiviral vector pLVX-AcGFP1-N1, the original vector backbone CMV promoter was removed; Cla I, Xho I double enzyme digestion full gene fragment, obtained radiation-induced promoter C 9 BC; will C 9 The BC promoter was ligated to the pLVX-AcGFP1-N1 fragment in which the CMV promoter was excised to obtain pLVX-C 9 BC-AcGFP1-N1;

[0049] (3) Using cDNA as a template to amplify the entire SOD2 gene fragment, the entire gene fragment contains the kozak sequence, the upstream of SOD2 contains the Xho I restriction site, and the downstream contains the BspT104I restriction site; Xho I, Bs...

Embodiment 2

[0079] Effect of irradiation treatment on the expression of SOD2 protein in stable cell lines

[0080] Given that 2-8Gy can effectively activate the chimeric promoter C 9 For the transcription of downstream genes by BC, experiments at the cell and animal levels were irradiated with γ-rays, with a total dose of 6Gy; animal experiments were irradiated with a single dose of 2Gy, once a day, for 3 consecutive days; experiments at the cell level were irradiated with a single dose of 6Gy method.

[0081] After observing 6Gy irradiation, the treatment vector (pLVX-C 9 BC-SOD2-T2A-AcGFP1) stable expression cell lines HT-29 and CCD 841 CoN expression changes of SOD2, combined with the previous part of the experimental results, 48h after irradiation treatment by western blot detection. The results showed that, compared with other groups, the expression of SOD2 in HT-29 stably transfected cell line was significantly increased after 6Gy irradiation ( Picture 1-1 and 1-2 ), the differe...

Embodiment 3

[0084] Effect of SOD2 overexpression on the proliferation ability of stable cell lines

[0085] The CCK-8 cell proliferation assay was used to detect the changes in cell proliferation ability among different cell groups. Experimental results such as image 3 shown. At 48h after irradiation, the cell proliferation ability of the SOD2+R group was lower than that of the Normal+R group, and the difference was significant (P<0.05); at 72h, the trend was more obvious, and the cell proliferation ability of the SOD2+R group was significantly lower than that of the Normal+R group ( P<0.01). It shows that the overexpression of SOD2 can increase the sensitivity of tumor cell HT-29 to radiation treatment and enhance the effect of radiation therapy.

[0086] Compared with the Normal+R group, the CCD 841 CoN stably transfected cell lines irradiated with 6Gy tended to improve the proliferation ability at 24h and 48h, but the difference was not significant. The cell proliferation ability ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a SOD2 lentiviral vector, a construction method and application thereof in targeted radiation protection and tumor sensitization and belongs to the technical field of tumor gene therapy. The vector is pLVX-C9BC-SOD2-T2A2-AcGFP1; an embedded promoter composed of a CAr G box and a CMV basal promoter in serial connection has obvious radiation inducing characteristics, can effectively promote expression of downstream gene under radiation treatment and can regulate and control the expression of the downstream gene within a certain range according to radiation dosage. In-vivo and in-vitro experiments prove that combined application of radiotherapy and SOD2 gene therapy has radiotherapy sensitization effect on tumor cells and can effectively inhibit proliferation and growth of the tumor cells; the combined application has radiation protection effect on normal cells and tissue and can lower apoptosis rate of the normal cells in a radiation field.

Description

technical field [0001] The invention relates to the technical field of tumor gene therapy, in particular to a SOD2 lentiviral vector, a construction method and its application in targeted radiation protection and tumor sensitization. Background technique [0002] Radiation therapy is one of the indispensable core technologies in the comprehensive treatment of tumors. In the local control of tumors, radiation will inevitably cause damage to normal tissues around the tumor. In order to reduce the damage of radiation to normal tissues, the dose and time of radiation will be strictly limited, which also restricts the therapeutic effect of radiotherapy to a certain extent. Multi-leaf collimators, precise fixation techniques, and radiation protection agents are often used to reduce radiation damage, but the effect is not ideal, and radiation protection agents lack targeting, and at the same time protect tumor tissues, thereby reducing the risk of radiation damage. The effect of r...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N15/867C12N15/66
CPCC12N15/66C12N15/86C12N2740/15043
Inventor 王卫东郎锦义张志强李宏阳章友刘晶郝玉徽李蓉王兴勇
Owner SICHUAN CANCER HOSPITAL
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com