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Oligodeoxyribonudeotide for preventing recurrence of tumor and inhibiting tumor growth as well as application thereof

An oligomeric nucleic acid, tumor technology, applied in anti-tumor drugs, introduction of foreign genetic material using vectors, gene therapy, etc., can solve problems such as the impact of tumor gene therapy, and achieve the effect of preventing tumor recurrence, long half-life, and inhibiting growth.

Active Publication Date: 2010-09-15
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In fact, more and more miRNAs have been identified as proto-oncogenes and tumor suppressor genes. Using the functions of miRNA tumor suppressor genes may have a great impact on tumor gene therapy, or may mimic this molecule for new medical research

Method used

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  • Oligodeoxyribonudeotide for preventing recurrence of tumor and inhibiting tumor growth as well as application thereof
  • Oligodeoxyribonudeotide for preventing recurrence of tumor and inhibiting tumor growth as well as application thereof
  • Oligodeoxyribonudeotide for preventing recurrence of tumor and inhibiting tumor growth as well as application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1, the preparation of oligomeric nucleic acid

[0021] The sense strand of the oligonucleotide sequence described in the present invention is: 5'-CUGUGCGUGUGACAGCGGCUGA-3' (SEQ ID No.1); the antisense strand is 5'-GACACGCACACUCGCCGACU-3' (SEQ ID No.2) or 5'- AGCCGCUGUCACACGCACAGUU-3' (SEQ ID No. 3). A random sequence was selected as a negative control (NC). The sense strand sequence of NC is: 5'-UUCUCCGAACGUGUCACGUTT-3' (SEQ ID No.4), and the antisense strand sequence is: 5'-ACGUGACACGUUCGGAGAATT-3' (SEQ ID No. 5). A, G, C, U in described oligonucleotide and negative control sequence represent adenine ribonucleotide, guanine ribonucleotide, cytosine ribonucleotide and uracil ribonucleotide, T represents thymus pyrimidine deoxynucleotides.

[0022] The oligonucleotide and the negative control commissioned Shanghai GenePharma Pharmaceutical Technology Co., Ltd. to synthesize and perform 2' fluoro (2'-F), 2' methoxy (2'-OMe), thio (PS) And 2' deoxy (2'-deoxy...

Embodiment 2、2

[0023] Effect of embodiment 2, 2'-F-oligonucleotide on tumor cell proliferation

[0024] The specific steps are: the 2'-F oligonucleotide obtained in Example 1 (the positive-sense strand is SEQ ID No1, and the antisense strand is SEQ ID No2) and the 2'-F oligonucleotide negative control powder are respectively dissolved in RNase-free Prepare a solution with a final concentration of 20 μmol / L in sterile water. Collect the tumor cells in the logarithmic phase, adjust the concentration of the cell suspension, and divide them into 96-well plates, 150 μl per well, a total of 5000 cells; store at 37 ° C, containing 5% CO 2 The cells were allowed to adhere to the wall in an incubator and cultured for 6-24 hours. For transfection, dilute 0.25 μl oligonucleotide solution (the above-mentioned 2′-F oligonucleotide or 2′-F oligonucleotide negative control) and 0.25 μl lipofectamine 2000 (Invitrogen) into 25 μl serum-free culture medium (Opti-MEM), and incubated at room temperature for 5...

Embodiment 3、2

[0025] Example 3, 2'-F-oligomeric nucleic acid inhibits the half-life test detection of tumor cell expression E2F3

[0026] Using Invitrogen's lipofectamine TM 2000 liposomes were used for transfection, and all operations were performed according to the operating procedures provided by Invitrogen. Inoculate tumor cells into 6-well plates (300,000 cells / well), culture with RPMI1640 or DMEM medium containing 10% fetal bovine serum for 16-24 hours, then replace the cell culture medium with RPMI1640 medium containing no antibiotics . For transfection, dilute 5 μl of oligonucleotide solution and 5 μl of lipofectamine 2000 in 250 μl of serum-free culture medium (Opti-MEM), and incubate at room temperature for 5 minutes, then mix the above oligonucleotide solution with lipofectamine 2000 The solutions were mixed, and after the complex was allowed to stand at room temperature for 20 minutes, 500 μl of the oligonucleotide-liposome complex was added to the cell culture plate (the cell...

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Abstract

The invention provides oligodeoxyribonudeotide for preventing tumor recurrence and inhibiting tumor growth as well as an application thereof. The oligodeoxyribonudeotide is in a double-strand structure and comprises a positive-sense strand and an antisense strand. The positive-sense strand has the sequence of 5'-CUGUGCGUGUGACAGCGGCUGA-3', the antisense strand has the sequence of 5'-GACACGCACACUCGCCGACU-3' or 5'-AGCCGCUGUCACACGCACAGUU-3', and the sequence can be chemically modified. The invention also provides a medicine composition containing the sequence and an application of the sequence to the preparation of medicines for preventing tumor recurrence caused by residual tumor cells after tumor extirpation or concurrent chemoradiotherapy and adult tumor growth.

Description

technical field [0001] The invention relates to a microRNA preparation for inhibiting tumor initiation and growth and its application. Background technique [0002] MicroRNAs (miRNAs) are a class of widely distributed non-coding small RNAs that regulate gene expression by partially complementing the 3′UTR (untranslated region) of the target sequence. In recent years, the correlation between miRNA-mediated post-transcriptional gene silencing and tumorigenesis has become a focus of attention. Studies have found that there are significant differences in miRNA expression profiles between tumor cells and normal tissue cells, and most miRNA genes are located at vulnerable gene loci associated with tumor formation, and miRNA genes appear at high frequencies in these variable genomic environments that are closely related to tumors , suggesting that there is a close relationship between miRNA in the occurrence, development and prognosis of tumors. In fact, more and more miRNAs have...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/63A61K48/00A61P35/00
Inventor 张雅鸥李建娜张佳荣胡祥张佩琢王均谢伟东何杰
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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