Recombinant lentiviral vector containing ubiquitin-specific protease gene USP39-shRNA (short hairpin ribonucleic acid) and application thereof

A technology of recombinant lentivirus and usp39-shrna, which is applied in the field of molecular biology, can solve the problems of breast cancer that have not been studied and the function of USP39 has not been elucidated.

Inactive Publication Date: 2013-07-17
HOSPITAL ATTACHED TO QINGDAO UNIV
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Problems solved by technology

[0004] Through the analysis of these existing studies, it is found that the function of USP39 is far from elucidated, and how it participates in the occurrence and development of breast cancer has not been studied so far.

Method used

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  • Recombinant lentiviral vector containing ubiquitin-specific protease gene USP39-shRNA (short hairpin ribonucleic acid) and application thereof
  • Recombinant lentiviral vector containing ubiquitin-specific protease gene USP39-shRNA (short hairpin ribonucleic acid) and application thereof
  • Recombinant lentiviral vector containing ubiquitin-specific protease gene USP39-shRNA (short hairpin ribonucleic acid) and application thereof

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Embodiment 1

[0032] The RNA extraction, reverse transcription and fluorescent quantitative reagents used in the present invention were purchased from Bao Bioengineering (Dalian) Co., Ltd., antibodies were purchased from Abcom, and various cell lines were purchased from ATCC (American Type Microorganism Collection Center) and Shanghai Cell Bank of Chinese Academy of Sciences (http: / / www.cellbank.org.cn / mulu.asp), various restriction endonucleases were purchased from NEB Company, the plasmids and gene chips used were purchased from Shanghai Jikai Biotechnology Company, and the plasmid extraction kit was purchased from Omega Company, Plus-20 centrifugal ultrafiltration device was purchased from Millipore Company, various biochemical reagents were purchased from Sigma Company, agarose was purchased from HydraGene Company, Alexa Fluor488annexin V / Dead Cell Apoptosis Kit, lipofectamine2000 were purchased from Invitrogen Company, ECL luminescence substrate The detection kit was purchased from Mill...

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Abstract

The invention provides a recombinant lentiviral vector containing ubiquitin-specific protease gene USP39-shRNA (short hairpin ribonucleic acid) and application thereof. The test proves that expression of USP39 in breast cancer tissue is obviously higher than that in normal breast tissue; the immunohistochemical detection confirms that the expression of USP39 in the breast cancer tissue is obviously higher than that in normal breast tissue; the immunohistochemical detection confirms that expression of USP39 protein in the breast cancer tissue is obviously higher than that in para-carcinoma tissue; siRNA (small interfering ribonucleic acid) is designed in a breast cancer cell system to interfere the expression of the USP39; it is found that proliferation of breast cancer cell after reducing the USP39 is obviously restrained; the cell apoptosis ratio is obviously improved; the cell ratio at the G1 stage is increased; the cell ratio at the S sage is reduced; and the clonality of the cell is obviously reduced. The test proves that the USP39 has an important role in facilitation of development of the breast cancer; and theoretical and experimental basis is provided for preparation of a drug for preventing and treating the breast cancer by the lentiviral vector containing USP39-shRNA.

Description

technical field [0001] The invention belongs to the field of molecular biology, in particular to recombinant lentivirus containing human ubiquitin-specific protease gene USP39-shRNA and application thereof. Background technique [0002] The ubiquitin-specific proteases (USPs) are the main members of the deubiquitinase family, which can recognize the specific sequence of ubiquitin and substrate, destroy the link between substrate protein and ubiquitin chain, play a role in deubiquitination. Deubiquitinases are involved in the regulation of cell cycle progression, the regulation of cell signal transduction, and the growth and development of organisms. More and more studies have found that the abnormal expression of USP family is closely related to various pathological changes, such as the occurrence of various tumors. Although more than 60 members of the USP family have been found in the human genome, the functions and mechanisms of most USPs are still unclear. [0003] The...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01A61K48/00A61P35/00C12R1/93
Inventor 王海波纪晓军刘相萍刘世海迟静薇
Owner HOSPITAL ATTACHED TO QINGDAO UNIV
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