Construction, screen and use for stomach cancer target AFP gene siRNAs expression carrier

An expression vector and gene technology, which can be used in the construction of siRNAs expression vector, gene therapy drugs, and screening fields, which can solve the problems of complex biological activity of AFP gene expression and incomplete elucidation.

Inactive Publication Date: 2007-04-04
汪运山
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Problems solved by technology

The increase of AFP content in adult blood is a sign of various diseases, including hepatocellular carcinoma, some gastrointestinal tumors and some germline embryonic tumors, etc. However,...

Method used

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  • Construction, screen and use for stomach cancer target AFP gene siRNAs expression carrier
  • Construction, screen and use for stomach cancer target AFP gene siRNAs expression carrier
  • Construction, screen and use for stomach cancer target AFP gene siRNAs expression carrier

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Embodiment Construction

[0077] The present invention will be further described below in conjunction with example, but not limited thereto.

[0078] 1. Main reagents

[0079] 1. Vector RNAi-Ready pSIREN-DNR-DsRed-Express Donor Vector was purchased from Clontech

[0080] 2. T4 DNA ligase and RT-PCR kit were purchased from Takara Company

[0081] 3. Plasmid Miniprep Kit QIAprep Spin Miniprep Kit was purchased from QIAGEN

[0082] 4. AFP-specific PCR primer synthesis, AFP-specific siRNA oligonucleotide chain synthesis and DNA sequence determination were completed by Shanghai Yingjun Bioengineering Technology Service Co., Ltd.

[0083] 5. Transfection reagents Lipofectamine 2000 Reagent and TRzol Reagent were purchased from Invitrogen

[0084] 6. The luciferase detection system was purchased from Promega

[0085] 7. Mouse anti-human α-fetoprotein monoclonal antibody, immunohistochemical staining kit and concentrated DAB chromogenic kit were purchased from Beijing Zhongshan Jinqiao Biotechnology Co., L...

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Abstract

Construction for constructing and screening siRNAs expression carrier of stomach cancer target AFP gene and its use are disclosed. Three expression plasmids in mammal cells by different AFP gene target sequences are constructed, and they are pSIREN-DNR-DsRed-Express Donor-Vector-AFP-siRNA1; pSIREN-DNR-DsRed-Express Donor Vector-AFP-siRNA2 and pSIREN-DNR-DsRed-express Donor Vector-AFP-siRNA3. It can inhibit AFP gene expression efficiently and be used to prepare gene medicine in treatment of stomach cancer.

Description

(1) Technical field [0001] The invention relates to a gene therapy drug for gastric cancer expressing an alpha-fetoprotein (alpha-fetoprotein, AFP) gene, in particular to the construction, screening and use of an siRNAs expression vector targeting the AFP gene, and belongs to the technical field of biomedicine. (2) Background technology [0002] As early as 1970, Bourreille J et al. published the article "Existence of alpha-fetoprotein in secondary liver cancer of gastric origin" in "Presse Med." (Bourreille J, Metayer P, Sauger F, et al. Existence of alpha-fetoprotein during gastric -origin secondary cancer of the liver. Presse Med. 1970 Jun 6; 78(28): 1277-8.), first reported a case of AFP positive gastric cancer (alpha-fetoprotein-producing gastric carcinoma, APGC). The literature reports on APGC mainly come from Japan, and the incidence of APGC in foreign literatures accounts for 5.1% to 15% of gastric cancer. The reported incidence rate in China is lower than that in f...

Claims

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Application Information

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IPC IPC(8): C12N15/63C12N15/11C07H21/02A61K48/00A61P35/00C12N15/113
Inventor 汪运山胡安拉马晓丽朱之炜肖东杰孙善会郏雁飞周芳
Owner 汪运山
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