Aquaporin 4-siRNA interfering sequence, its fusion expression vectors and medicinal use of the fusion expression vectors

An aquaporin and interfering sequence technology, applied in the field of aquaporin 4-siRNA interfering sequence and its fusion expression vector, can solve the problems that are not yet fully clarified, and achieve the effects of inhibiting invasion and infiltration and inhibiting tumor spread

Inactive Publication Date: 2011-11-16
TIANJIN MEDICAL UNIV CANCER HOSPITAL
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Problems solved by technology

So far, the molecular mechanism of invasion and inva

Method used

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  • Aquaporin 4-siRNA interfering sequence, its fusion expression vectors and medicinal use of the fusion expression vectors
  • Aquaporin 4-siRNA interfering sequence, its fusion expression vectors and medicinal use of the fusion expression vectors
  • Aquaporin 4-siRNA interfering sequence, its fusion expression vectors and medicinal use of the fusion expression vectors

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[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 AQP4-siRNA

[0029] Design specific intersectinl-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: 5'-GGCTGGCTTGGAGGAGAATTA-3'.

[0030] 3. Preparation of AQP4-siRNA fusion expression vector

[0031] 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 AQP4-siRNA was annealed, connected into the pGPU6 / GFP / Neo...

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Abstract

The invention discloses an aquaporin 4-siRNA interfering sequence, its fusion expression vectors and a medicinal use of the fusion expression vectors, and belongs to the biopharmaceutical field. In the invention, an aquaporin 4-siRNA interfering sequence is designed and fusion expression vectors of the aquaporin 4-siRNA interfering sequence are synthesized. After the fusion expression vectors are transfected into human glioma cell lines, the fusion expression vectors can degrade specifically aquaporin 4mRNA which is complementary with the fusion expression vectors, decrease the expression of an aquaporin protein 4 in a human glioma cell, inhibit effectively a migration capacity, an adhesive capacity and an invasion capacity of human glioma cells and have effects of inhibiting the infiltration and the diffusion of human glioma cells thus provide a feasible technical means for a gene therapy utilized for tumor patients.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an aquaporin 4-siRNA interference sequence for inhibiting the infiltration and spread of glioma cells, 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 Research 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 single injections of sense and antisense strands. This suppression of specific gene expression triggered by dsRNA is called RNA interference (RNA interferenee RNAi). [0003] RNA interference is sequence-specific gene silencing caused by double-...

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Application Information

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IPC IPC(8): C12N15/113C12N15/63A61K48/00A61P35/00A61P35/04A61P25/00
Inventor 谷峰马勇杰丁婷
Owner TIANJIN MEDICAL UNIV CANCER HOSPITAL
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