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Human miR-150 antisense nucleic acid and application thereof

An antisense, nucleotide technology, applied in the application field of antisense oligonucleotides, can solve the problems of poor curative effect, insignificant improvement in survival rate of tumor patients, no significant improvement, etc., and achieve inhibition of growth and malignant proliferation. effect of ability

Active Publication Date: 2011-04-27
SUZHOU GENEPHARMA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] 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, hovering At about 30% to 55%, there is no significant improvement, and the 5-year survival rate of middle and advanced patients is even lower, about 20%.
Moreover, these methods have their own limitations, especially for middle-advanced and relapsed patients.

Method used

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  • Human miR-150 antisense nucleic acid and application thereof
  • Human miR-150 antisense nucleic acid and application thereof
  • Human miR-150 antisense nucleic acid and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1, miR-150 antisense nucleic acid inhibits the expression of miR150

[0031] The real-time quantitative fluorescence detection test was completed by Shanghai Gemma Pharmaceutical Technology Co., Ltd. The specific experimental steps include: cell culture: U87 cells, 10% FBS-DMEM medium culture, 37 ° C, 5% CO2 culture.

[0032] Cell transfection:

[0033] 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%;

[0034] 2) Transfection samples were prepared as follows to prepare oligomers-Lipofecta mine TM 2000 complex:

[0035] a. Use 50 μl serum-free Opti- Dilute miR-150 antisense oligonucleotide (sequence 5'-CACUGGUACAAGGGUUGGGAGA-3'), negative control, and FAM-labeled negative control in I culture medium respectively, the final concentration is 50nM, mix gently, each transfection Set up 3 multiple...

Embodiment 2

[0065] Example 2, Detection of inhibitory activity of MiRNA antisense oligonucleotides on human glioblastoma cell line U87

[0066] Cell culture:

[0067] U87 cells were cultured in 10% FBS-DMEM medium, 37°C, 5% CO2. Collect U87 cells in good growth state, count by centrifugation, and use 2×10 3 Spread each well in a 96-well plate and culture at 37°C with 5% CO2.

[0068] Transfection:

[0069] 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%;

[0070] 2) Transfection samples were prepared as follows to prepare oligomers-Lipofecta mine TM 2000 complex:

[0071] a. Use 25 μl serum-free Opti- Dilute the miR-150 antisense oligonucleotide, negative control, and FAM-labeled negative control in I culture medium respectively to a final concentration of 50 nM, mix gently, and set 3 duplicate holes for each transf...

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Abstract

The invention discloses antisense oligonucleotide for suppressing the expression of micro ribonucleic acid (RNA)-150 and application thereof. The antisense oligonucleotide comprises a sequence, namely, 5'-CACUGGUACAAGGGUUGGGAGA-3', wherein the sequence can be specifically combined with human miR-150. The antisense oligonucleotide can be ribonucleotide, deoxyribonucleotide or the chimera of the ribonucleotide and the deoxyribonucleotide and can modify any nucleotide in a chain. MiR-150 antisense oligonucleotide can effectively suppress the expression of miR-150 in a human glioma cell U87, suppresses the growth and multiplication of the cell and can effectively treat brain glioma and other tumors with high miR-150 expression.

Description

technical field [0001] The invention relates to the field of biomedicine. Specifically, the present invention relates to the use of a small molecule microRNA (miRNA), especially the application of antisense oligonucleotides related to the miRNA. The antisense oligonucleotide can be complementary to human miR-150, thereby inhibiting the expression of human miR-150. The present invention also relates to a pharmaceutical composition containing the miRNA antisense oligonucleotide. Background technique [0002] miRNAs are small non-coding RNAs with a length of 20-22bp, usually transcribed by RNA polymerase II (PolII), and generally the initial product is a large one with a cap structure (7MGpppG) and a polyA tail (AAAAA) 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 this precursor molecule into the cytoplasm. Subsequently, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11A61K48/00A61P35/00
Inventor 丁侃张佩琢东楠李捷沈孝坤
Owner SUZHOU GENEPHARMA
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