Cytoskeletal binding protein siRNA (small interfering ribonucleic acid) interfering sequence, fusion expression vector thereof and medical application of same

A technology of actin and protein binding, applied in DNA/RNA fragments, pharmaceutical formulations, recombinant DNA technology, etc., can solve problems that have not been fully clarified, and achieve the effect of inhibiting tumor spread, invasion and infiltration

Active Publication Date: 2013-01-02
TIANJIN MEDICAL UNIV CANCER HOSPITAL
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Problems solved by technology

So far, the molecular mechanism of invasion and invasion of malignant glioma has not been fully clarified

Method used

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  • Cytoskeletal binding protein siRNA (small interfering ribonucleic acid) interfering sequence, fusion expression vector thereof and medical application of same
  • Cytoskeletal binding protein siRNA (small interfering ribonucleic acid) interfering sequence, fusion expression vector thereof and medical application of same
  • Cytoskeletal binding protein siRNA (small interfering ribonucleic acid) interfering sequence, fusion expression vector thereof and medical application of same

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Experimental program
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Embodiment Construction

[0026] 1. Sources of experimental materials:

[0027] Human malignant glioma cell line LN-229 was provided by the University of Texas MD Anderson Cancer Center, the chemotaxis chamber was purchased from Neuro Probe, EGF was purchased from Peprotech, the intersectin1 antibody was from Abcam, and G418 was purchased from Invitrogen company.

[0028] 2. Design and synthesis of Girdin-siRNA

[0029] Design specific intersectin1-siRNA according to the siRNA design software provided by Ambion on the Internet, and synthesize the following templates by Shanghai Gemma Pharmaceutical Technology Co., Ltd., with BamH I and Bbs I restriction sites at both ends, and siRNA action sites:

[0030] 5'-GAAGGAGAGGCAACTGGAT - 3'.

[0031] 3. Preparation of Girdin-siRNA fusion expression vector

[0032] The expression vector pGPU6 / GFP / Neo vector has a linear structure, with sticky ends of BamH I and Bbs I restriction sites at both ends. The synthesized Girdin-siRNA was annealed, connected into t...

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Abstract

The invention discloses a cytoskeletal binding protein siRNA (small interfering ribonucleic acid) interfering sequence, a fusion expression vector thereof and medical application of the same, belonging to the field of biological pharmacy. The invention designs a segment of cytoskeletal binding protein siRNA interfering sequence, and synthesizes the fusion expression vector of the cytoskeletal binding protein siRNA interfering sequence; after the fusion expression vector is transfected into a human glioma cell line, the fusion expression vector can specifically degrade the cytoskeletal binding protein messenger RNA (mRNA) complementary to the fusion expression vector to reduce the expression of the cytoskeletal binding protein in the human glioma cells; and the fusion expression vector can effectively inhibit the migration, adhesion and invasion abilities of the human glioma cells, has an effect on inhibiting the human glioma cellular infiltration and metastasis, and provides a feasible technical means for gene therapy for tumor patients.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a cell actin binding protein siRNA interference fragment for inhibiting glioma cell infiltration and spread, its fusion expression vector and the medical application of the expression vector. Background technique [0002] Post-transcriptional gene silencing is an evolutionary defense mechanism against transcriptomes and RNA viruses that arose before the divergence of plants and animals. In 1998, Andrew Fire of the Carnegie Institute of Washington and Craig Mello of the University of Massachusetts Cancer Center injected double-stranded dsRNA (double-stranded RNA) into nematodes for the first time, which induced gene silencing stronger than individual injections of the sense and antisense strands. This suppression of specific gene expression triggered by dsRNA is called RNA interference (RNA interference RNAi). [0003] RNA interference is fragment-specific gene silencing caused by do...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/85A61K48/00A61P35/00A61P35/04
Inventor 马勇杰谷峰王静
Owner TIANJIN MEDICAL UNIV CANCER HOSPITAL
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