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Attenuated salmonella transporting recombinant plasmid and its application in treating tumor

A recombinant plasmid, Salmonella technology, used in anti-tumor drugs, applications, recombinant DNA technology, etc., to prolong survival time, inhibit tumor growth and metastasis.

Active Publication Date: 2007-06-06
吉林启众生物科技有限公司
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AI Technical Summary

Problems solved by technology

[0006] Our previous work has successfully established a recombinant plasmid for the treatment of prostate cancer and melanoma, and applied for a patent, patent application number: CN200510017056.9; publication number: CN1730658, currently carrying siRNA-Stat3 in attenuated Salmonella, siRNA-Survivin and the pGRIM-19-Si-Stat3 recombinant plasmid co-expressing siRNA-Stat3 and GRIM-19 are used to treat a variety of tumors, which have not been reported at home and abroad

Method used

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  • Attenuated salmonella transporting recombinant plasmid and its application in treating tumor
  • Attenuated salmonella transporting recombinant plasmid and its application in treating tumor
  • Attenuated salmonella transporting recombinant plasmid and its application in treating tumor

Examples

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preparation example Construction

[0158] 2.1 Preparation of electroconversion competent state

[0159] Pick up the monoclonal attenuated Salmonella colony with a pipette tip, and put it into a 50ml centrifuge tube filled with 5ml LB liquid medium. (Blank control of culture medium and tip at the same time) 37°C, 220rpm, cultivate for 14-16 hours. The next day, pour the 5ml bacterial solution into 500ml LB liquid medium at a ratio of 1:100, shake at 37°C and 220rpm for 2-3h, measure the OD value every half hour, when the OD value reaches 0.3-0.4 , stop culturing. The bacterial solution was pre-cooled on ice for 30 minutes, and then the bacterial solution was distributed into 100ml pre-cooled centrifuge tubes, and centrifuged at 4200rpm for 10min at 4°C. Discard the supernatant, add 1mmol / L ice-cooled HEPES (pH7.0) 1ml to the centrifuge tube to resuspend the pellet, then add 1mmol / L ice-cooled HEPES (pH7.0) 49ml, 4°C, 4200rpm Centrifuge for 10 min. Repeat 2 times. Discard the supernatant, add a small amount ...

Embodiment

[0239] Formulate the invention with PDS as 3 x 10 8 / ml solution into 1ml ampoule. When in use, inject 1ml subcutaneously or intramuscularly per person.

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PUM

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Abstract

The present invention relates to attenuated salmonella transporting recombinant plasmid and its application in treating tumor, and belongs to the field of biotechnology. The present invention constitutes pH1Si-Stat3, pGRIM-19 and pGRIM-19-Si-Stat3 recombinant plasmid capable of expressing GRIM-19 and siRNA-Stat3 simultaneously, and translates the recombinant plasmid electrically into attenuated salmonella. Intracorporeal experiment proves that the coexpression of both GRIM-19 and siRNA-Stat3 results in excellent synergistic effect of inhibiting tumor growth and metastasis. Attenuated salmonella possesses tumor phagocytising characteristic and can propagate in tumor cell in the specificity to that in health cell over 1000 times, so that attenuated salmonella carrying transporting recombinant plasmid capable of expressing GRIM-19 and siRNA-Stat3 simultaneously to the depth of tumor can inhibit tumor growth and metastasis obviously.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to attenuated Salmonella carrying siRNA-Stat3, siRNA-Survivin and co-expressing siRNA-Stat3 and GRIM-19 plasmids, a preparation method thereof and an application in treating tumors. Background technique [0002] One of the main obstacles in tumor gene therapy is how to deliver anti-tumor factors or other therapeutic drugs into tumor tissues, especially deep tumors and metastatic tumors, without harming normal tissues. Salmonella is a facultative anaerobic bacterium with tumor-tropic activity. It can reproduce well in the deep part of the tumor under hypoxic or hypoxic conditions and produce oncolytic effects. Many reports have pointed out that the specificity of Salmonella in tumor tissue is compared with normal tissue. It can be more than 1000 times, so the use of attenuated Salmonella in tumor treatment has a very promising application prospect. [0003] The oncogene Stat3...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/09C12N15/12A61K35/74A61P35/00C12R1/42C12N15/11C12N15/113
CPCA61K38/00A61K35/74C12N2320/32C12N2310/111C12N1/36C12N2310/14C12N15/111C07K14/4747C12N15/1135A61P35/00Y02A50/30
Inventor 张灵赵雪俭徐德启高丽芳
Owner 吉林启众生物科技有限公司
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