Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Gene therapeutic drug hdm2-siRNA for breast cancer

An anti-breast cancer and drug technology, applied in gene therapy, drug combination, sugar derivatives, etc., can solve the problems of slow progress in applied research, high dosage, and insufficient specificity, and achieve the effect of wide application prospects

Inactive Publication Date: 2006-03-29
MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the limitations of antisense nucleic acid itself, its specificity is not strong enough and the dosage is too high, the progress of further application research is slow.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Gene therapeutic drug hdm2-siRNA for breast cancer
  • Gene therapeutic drug hdm2-siRNA for breast cancer
  • Gene therapeutic drug hdm2-siRNA for breast cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] siRNA target sequence selection

[0037] First, it may be more effective to select a 21-nucleotide target region from the sequence between 50 nucleotides downstream of the start codon and 100 nucleotides before the stop codon in the complete cDNA sequence of a specific gene (Genes and Development 2001, 15: 188-200). Since there are many regulatory protein binding sites in the 5' or 3' untranslated region and near the start codon, these sites are generally not selected. Binding proteins in the untranslated region and translation initiation complexes may affect the binding of short interfering ribonucleoprotein (short interfering Ribonucleoprotein, siRNP) and RNA-induced silencing complex (RISC) to target mRNA. The sequence used is generally a 21 nucleotide sequence whose first two are AA and the last two are TT, and the G / C content is 30%-50%. If no such sequence is contained in the cDNA, a 21 nucleotide sequence with only the first two nucleotides being AA can also be...

Embodiment 2

[0057] siRNA transfection

[0058] The present invention uses LipofectAMINE TM 2000 liposome (Life Technologies, product number: 11668-027) was used for transfection, and the silencing effect was measured 18-45 hours after transfection. The synthesized siRNA powder was dissolved in water to prepare a 4000 nM stock solution. 18-20 hours before transfection, the experimental cells were inoculated into 96-well plates at 6000-8000 cells / well in 200 μl of antibiotic-free DMEM tissue culture medium containing 10% fetal bovine serum. Dilute 4000nM siRNA to 75μl each of 120nM, 400nM, 1200nM, 2000nM, and 4000nM with Opti-MEM medium without antibiotics and serum. In another Eppendorf tube, add 5μl of liposomes to 370μl of Opti-MEM medium After incubation for 5 minutes, add 75 μl liposome diluent to each concentration of siRNA solution and mix gently. After the siRNA-liposome complex was allowed to stand at room temperature for 20 minutes, 50 μl of the siRNA-liposome complex was added...

Embodiment 3

[0060] Detecting the Inhibitory Effect of siRNA on Tumor Cell Growth by MTT Assay

[0061] Cells transfected according to the method of Example 2 were stored at 37°C, containing 5% CO 2 Incubate for about 45 hours in an incubator with air and 100% humidity. MTT was made into a 2 mg / ml solution with PBS buffer, 50 μl was added to each well, and incubated at 37°C for 4 hours to reduce MTT to Formazan. The supernatant was aspirated and 150 μl of DMSO (dimethyl sulfoxide) was added to dissolve Formazan. Measure the optical density OD of each well at 570 nm with a microplate reader (Mode13550, Bio-Rad) 570. The OD value of each test well is subtracted from the background OD value (complete medium plus MTT, no cells) or the OD value of the blank drug well (complete medium plus different dilutions of the test drug plus MTT, no cells), and each repeat The OD values ​​of the wells were taken as mean ± SD. The survival rate of the cells is represented by T / C%, T is the OD value of ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a breast-cancer-treating genetic drug hdm2-siRNA design, synthesizing, cytosis detecting, target mRNA and target protein level silence action detection, induced cell apoptosis and cycle retardation detection and in vivo antineoplastic action detection, wherein the short interference RNA (siRNA) designed aiming hdm2 gene can inhibit the accretion or MCF-cell after cell transfection, specifically lower the mRNA level and HDM2 protein expression level of the hdm2 genes, and induce MCF-7 cytogenesis apoptosis and G1 period retardation simultaneously.

Description

Technical field: [0001] The invention relates to the design and synthesis of a short interfering ribonucleic acid (short interfering RNA, siRNA) drug and the detection of its growth inhibitory effect, apoptosis induction effect, cycle arrest effect and in vivo anti-tumor effect. Background technique: [0002] With the increasingly serious environmental pollution and changes in other living conditions, the mortality rate of breast cancer patients has gradually increased in recent years, and 500,000 to 1,000,000 people worldwide die of breast cancer every year. In 2003, the mortality rate of breast cancer in the United States was second only to skin cancer among female malignancies (CA Cancer J Clin. 2003, 53(1): 5-26), while its mortality rate in Beijing has become the first among female malignancies. Therefore, research on gene therapy for breast cancer has certain practical significance. [0003] Breast cancer is regulated by a network of multiple genes. The hdm2 gene is a...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C07H21/00A61K48/00A61P35/00C07H21/02
Inventor 邵荣光刘铁刚殷勤伟何红伟
Owner MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com