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Antisense STAT3 oligonucleotide and uses thereof

An oligonucleotide and antisense technology, applied in the field of high-efficiency antisense STAT3 oligonucleotide and its application in anti-tumor drugs, can solve the problems of unsatisfactory anti-tumor effect, and improve radiation sensitivity Effect

Inactive Publication Date: 2008-01-30
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current antisense nucleic acid drugs also have shortcomings. Due to the different targets selected, their anti-tumor effect is not very satisfactory, and there is still a long way to go before practical application.

Method used

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  • Antisense STAT3 oligonucleotide and uses thereof
  • Antisense STAT3 oligonucleotide and uses thereof
  • Antisense STAT3 oligonucleotide and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Prediction of the secondary structure of STAT3 mRNA

[0026] Using the mRNA secondary structure prediction software RNAstructure 4.2 (for Windows version), the full-length sequence of STAT3 mRNA (sequence number: L29277) was retrieved from the gene library genebank, and the secondary structure of STAT3 mRNA was predicted by the software, and the predicted secondary structure of STAT3 mRNA was obtained. structure, on the basis of the predicted secondary structure, use the oligowalk module of the software to design antisense sequences with relatively low total free energy, a total of 10, the minimum free energy of each sequence is shown in Table 1, each sequence crosses Synthesized by Shanghai Sangong, modified with full thio, purified with polyacrylamide.

[0027] Table 1 STAT3 antisense nucleic acid sequence oligowalk program designed with RNAstructure 4.2 simulation software oligowalk module

[0028] No.

Embodiment 2

[0029] Example 2: Inhibitory effect of each sequence on the proliferation of lung gland A549 cells

[0030] Positive control antisense sequence (ASC-STAT3) reported in the literature: 5'-GCTCCAGCATCTGCTGCTTC-3', nonsense sequence (NS-STAT3): 5'-GCTCCAGCATCTGCTGCTTC-3', each sequence pair was detected by CCK-8 assay Proliferation inhibitory effect of lung adenocarcinoma A549 cells (American Type Culture Collection, available from ATCC). Its steps include:

[0031]2.1 Cell culture A549 cells were cultured with RPMI-1640 medium containing 10% fetal bovine serum, 100 u / ml penicillin, and 100 μg / ml streptomycin at 37°C and 5% CO 2 Cultured under saturated humidity conditions, the cells used in the experiment were in the exponential growth phase.

[0032] 2.2 Oligonucleotide transfection Each oligonucleotide was encapsulated with cationic liposomes (Oligofectamine purchased from Invitrogen) to transfect A549 cells, and the operation was performed according to the kit instructions....

Embodiment 3

[0036] Example 3: Inhibitory effect of different concentrations of AS10 on the proliferation of lung adenocarcinoma A549 cells

[0037] Then use the optimal sequence group AS10, the positive control group (ASC), and the nonsense group (NS) to transfect in a 96-well plate at different concentrations (50, 100, 150, 200, 250, 300, 400nmol / L), After 48 hours, the above-mentioned method was used to calculate the survival fractions of each concentration in each group, and the results are shown in Fig. 2 .

[0038] Survival fraction=(administration group-background) / (normal control group-background)×100%.

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Abstract

The invention pertains to biological engineering field and relates to antisense ribonucleotide technology, in particular to antisense STAT3 oligonucleotide and the application thereof in antitumor drugs. The STAT3 becomes one of the hot study and application issues in the world because the STAT3 is a promising molecular target in the phyma treatment field. The invention discloses three series of high-effective antisense STAT3 oligonucleotide series which has proliferation inhibition effect to phyma cells, and compared with the known series, proliferation inhibition effect to phyma cells is improved and apoptosis of phyma cells can be effectively induced; therefore, the invention can be applied to the preparation of antitumor drugs. Combined with radiation factors, the invention is observed to evidently reduce radiation sensitivity of phyma cell and the invention can be used as radiation sensitizing agent.

Description

technical field [0001] The invention belongs to the field of bioengineering and relates to antisense nucleic acid technology, in particular to high-efficiency antisense STAT3 oligonucleotide and its application in antitumor drugs. Background technique [0002] Antisense nucleic acid technology is a new technology in the field of molecular biology after gene cloning and recombination technology. It uses biosynthesized or artificially synthesized complementary antisense nucleic acids or their chemical modifications to Specific nucleic acids within cells bind to inhibit or block their expression, thereby interfering with the production of disease-causing proteins. In terms of tumor treatment, people have designed many anti-tumor antisense nucleic acids targeting different mutated genes in the process of carcinogenesis, and conducted extensive research, some of which have entered the stage of clinical trials. [0003] At present, antisense nucleic acid drugs in the world have a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C07H21/04A61K31/7088A61K48/00A61P35/00C12N15/113
Inventor 蔡建明祝宝让唐古生高福李百龙
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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