Structure and application of siRNA for suppressing proliferation and metastasis of human lung cancer cells

A technology of vascular endothelium and growth factors, applied in DNA/RNA fragments, recombinant DNA technology, gene therapy, etc., can solve the problem of low survival rate and achieve important social and economic benefits

Inactive Publication Date: 2011-01-12
江其生
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Problems solved by technology

Although a variety of treatment methods are used, the 3-year and 5-year survival rates of patients are s

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  • Structure and application of siRNA for suppressing proliferation and metastasis of human lung cancer cells

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Embodiment Construction

[0015] The experimental design, methods and experimental results are as follows:

[0016] 1. siRNA design and synthesis and plasmid vector construction

[0017] The full-length sequence of human VEGF mRNA (sequence number: NM 001033756, CDS: 492-1607) was obtained from the NCBI database of the National Center for Biotechnology Information, USA. Based on the principles of siRNA design, combined with design software and literature reports, 12 VEGF-siRNA target sequences were designed. The length of each siRNA target sequence is 19bp, and all BLAST comparison checks are performed to ensure that there is no homology with other genes. The length of the synthesized siRNA transcription template is 64bp, and a forward and reverse target sequence separated by a 9bp stem-loop sequence is added in the middle of the sequence. 5 Ts were inserted at the end as a termination sequence. BamHI and Hind11I restriction sites were added upstream and downstream, respectively. Plasmid (digested w...

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Abstract

The invention provides a structure and application of small interference RNA (siRNA) for suppressing proliferation and metastasis of human lung cancer cells, and relates to the bioengineering pharmaceutical field, in particular to a method for designing, synthesizing and screening the small interference RNA (siRNA) for suppressing (silencing) gene expression of human VEGF so as to suppress the proliferation, invasion and metastasis of malignant cells such as lung cancer cells and the like according to gene sequence information on a vascular endothelial growth factor (VEGF). Through the structure and the application, that the siRNA which is complementary with a specified region of the VEGF mRNA has a suppression effect on the proliferation and the metastasis of the human lung cancer A549 cells cultured in vitro is designed and discovered. Thus, the invention relates to the siRNA sequence, the structure and a novel medicament thereof for preventing and curing the proliferation and the metastasis of the malignant cells such as lung cancer cells and the like aiming at the VEGF gene.

Description

technical field [0001] The present invention relates to the field of bioengineering drugs, in particular to a sequence structure of small interfering RNA (small interference RNA, siRNA) targeting vascular endothelial growth factor (vascular endothelial growth factor, VEGF) gene and its potential for treating malignant tumor cells such as lung cancer. New drugs for proliferation, invasion and metastasis. Background technique [0002] Lung cancer has become one of the common diseases causing human death, and its morbidity and mortality rate ranks first in various malignant tumors in large and medium-sized cities in my country. Although a variety of treatment methods are used, the 3-year and 5-year survival rates of patients are still very low, and the main cause of patient death is directly related to tumor recurrence and metastasis. [0003] The proliferation, invasion and metastasis of cancer cells is a complex process regulated by multiple factors. The proliferation of ca...

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Application Information

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IPC IPC(8): C12N15/113A61K48/00A61P11/00A61P35/00A61P35/04
Inventor 江其生
Owner 江其生
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