Human miR-145 antisense nucleic acid and application thereof

An antisense and oligonucleotide technology, applied in DNA/RNA fragments, gene therapy, anti-tumor drugs, etc., can solve the problem of poor curative effect of patients with distant metastasis, low survival rate of middle and advanced patients, and low survival rate and other problems, to achieve the effect of inhibiting growth and malignant proliferation

Inactive Publication Date: 2012-03-21
SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] In the past 30 years, although the comprehensive treatment of tumors has been very common clinically, the comprehensive treatment based on surgery and supplemented by radiotherapy and chemotherapy has not significantly improved the survival rate of cancer patients, and the 5-year overall survival rate is still low, hoveri

Method used

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  • Human miR-145 antisense nucleic acid and application thereof
  • Human miR-145 antisense nucleic acid and application thereof
  • Human miR-145 antisense nucleic acid and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1, miR-145 antisense nucleic acid inhibits the expression of miR-145

[0035] For real-time quantitative fluorescence detection, the oligonucleotide sequence involved was synthesized by Shanghai Gemma Pharmaceutical Technology Co., Ltd. The specific experimental steps include:

[0036] Cell culture: U87 / MG cells (purchased from the Cell Bank of the Type Culture Collection Committee of the Chinese Academy of Sciences), cultured in 10% FBS-DMEM medium (FBS was purchased from Hyclone, DMEM was purchased from Gibco), 37 ° C, 5% CO 2 to cultivate.

[0037] Cell transfection:

[0038] 1) One day before transfection, inoculate cultured cells in a 24-well plate with an appropriate amount of culture medium without antibiotics, so that the confluence of cells at the time of transfection reaches 30-50%;

[0039] 2) Transfection samples were prepared as follows to prepare oligomers-Lipofecta mine TM 2000 complex:

[0040] a. Use 50 μl serum-free Dilute miR-145 antisen...

Embodiment 2

[0072] Example 2. Detection of inhibitory activity of miR-145 antisense oligonucleotides on human glioma cell line U87 / MG

[0073] The oligonucleotide sequences involved were synthesized by Shanghai Gemma Pharmaceutical Technology Co., Ltd.

[0074] Cell culture:

[0075] U87 / MG cells (purchased from the Cell Bank of the Type Culture Collection Committee of the Chinese Academy of Sciences), cultured in 10% FBS-DMEM medium (FBS was purchased from Gibco, DMEM was purchased from Hyclone), 37 ° C, 5% CO 2 to cultivate. Collect U87 / MG cells in good growth state, count by centrifugation, and use 2×10 3 Spread each well in a 96-well plate at 37°C, 5% CO 2 Cultivate for 24h.

[0076] Transfection:

[0077] 1) One day before transfection, inoculate cultured cells in a 96-well plate with an appropriate amount of culture medium without antibiotics, so that the confluence of cells at the time of transfection reaches 30-50%;

[0078] 2) Transfection samples were prepared as follows t...

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Abstract

The invention discloses an antisense oligonucleotide for suppressing the expression of microRNA (ribose nucleic acid)-145 and application of the antisense oligonucleotide. The antisense oligonucleotide is specifically combined to human miR-145 and comprises a sequence complementary with at least thirteen continuous nucleotides in the nucleotide sequence of 5'-GUCCAGUUUUCCCAGGAAUCCCU-3', and particularly comprises the sequence of 5'-AGGGAUUCCUGGGAAAACUGGAC-3'. The antisense oligonucleotide can be a ribonucleotide, a deoxyribonucleotide or a chimera of the ribonucleotide with the deoxyribonucleotide, and an optional nucleotide in a chain can be modified. The miR-145 antisense oligonucleotide is capable of effectively suppressing the expression of miR-145 in human cerebral glioma cells, and suppressing growth and proliferation of the miR-145, thereby effectively treating cerebral gliomas and other miR-145 highly-expressed tumors.

Description

technical field [0001] The invention relates to the field of biomedicine. Specifically, the present invention relates to a use of microRNA (miRNA), in particular to an antisense oligonucleotide for inhibiting the expression of human microRNA-145 (miR-145) and its application. The antisense oligonucleotide can be complementary to human miR-145, thereby inhibiting the expression of human miR-145 to play an anti-tumor effect. The present invention also relates to a pharmaceutical composition containing the miRNA antisense oligonucleotide. Background technique [0002] miRNAs are small non-coding RNAs, 20-25bp in length, usually transcribed by RNA polymerase II (Pol II), and generally the initial product is a large one with a cap structure (7MGpppG) and a polyA tail (AAAAA) The pri-miRNA. These pri-miRNAs are processed into pre-miRNA precursor products consisting of 70 nucleotides under the action of RNase III Drosha and its cofactor Pasha. RAN-GTP and exportin 5 transport t...

Claims

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Application Information

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IPC IPC(8): C12N15/113A61K48/00A61P35/00A61P35/02
Inventor 丁侃张佩琢李捷东楠沈孝坤
Owner SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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