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

A method for regulating the expression, quantity and activity of heat shock protein 70 and its application

An active and protein technology, applied in the direction of peptide/protein components, medical preparations containing active ingredients, pharmaceutical formulations, etc., can solve the problems of limited stability and solubility, limited clinical application, low bioavailability, etc., to achieve inhibition of the entity Tumor growth, enhanced activation, and sensitivity-enhancing effects

Active Publication Date: 2018-02-02
NANJING UNIV
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the drugs geldanamycin and 17-AAG targeting HSP90 as therapeutic targets have entered clinical research. However, although geldanamycin and 17-AAG have good anti-tumor activity, they still have limitations, mainly in the stability and solubility. Large, can be metabolized by cytochrome P450, low bioavailability, and geldanamycin has severe liver toxicity, which limits its clinical application

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
  • A method for regulating the expression, quantity and activity of heat shock protein 70 and its application
  • A method for regulating the expression, quantity and activity of heat shock protein 70 and its application
  • A method for regulating the expression, quantity and activity of heat shock protein 70 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Using RNA interference technology to down-regulate the expression of HSP70 in tumor cells or tissues, combined with the anti-tumor drug TRAIL to induce tumor cell apoptosis, and treat the mouse A549 solid tumor model as an example:

[0073] 1. Construction of HSP70 interference plasmid:

[0074] Genscript's software siRNA target finder was used to select the appropriate target sequence from the HSP70 sequence, and then Genscript's software siRNA Construct Builder was used to construct a DNA fragment for interference that could be connected to the vector pRNAT-U6.1 / Neo. The DNA fragment includes a sense strand (sense strand, which is the same as the target sequence), a hairpin structure, an antisense strand (antisense strand, which is complementary to the sense strand), and a poly T signal at the 3' end that can terminate transcription; in addition, in order to facilitate It was connected to the vector pRNAT-U6.1 / Neo, and restriction sites for BamH I (G↓GATCC) and Hind I...

Embodiment 2

[0121] Using RNA interference technology to down-regulate the expression of HSP27 in tumor cells or tissues, combined with the anti-tumor drug TRAIL to induce tumor cell apoptosis, and treat the mouse A549 solid tumor model as an example:

[0122] 1. Construction of HSP27 interference plasmid:

[0123] The design and construction method of the HSP27 interference plasmid is the same as that described in Example 1. The fragment for HSP27 RNA interference designed by Gensicript software is as follows: 5'-GGATCCCGTCTCATCGGATTTTGCAGTTGATATCCGCTGCAAAATCCGATGAGACTTTTTTCCAAAAGC-3', and the two fragments synthesized by Shenergy Gambling Biotechnology Co., Ltd. are: 5'-GATCCCGTCTCATCGGATTTTGCAGTTGATATCCGCTGCAAAATCCGATGAGACTTTTTTCCAAA-3' -AGCTTTTGGAAAAAAGTCTCATCGGATTTTGCAGCGGATATCAACTGCAAAATCCGATGAGACGG-3'.

[0124] 2. Detection of the interference effect of HSP27 interference plasmid and its ability to induce A549 cell apoptosis at the cellular level when combined with TRAIL:

[0125]...

Embodiment 3

[0154] Detection of the ability of HSP70 interference combined with TRAIL to induce apoptosis of various non-small cell lung cancer cells NSCLC (A549, SW1573 and H460) at the cellular level (cell apoptosis detected by flow cytometry):

[0155] Transfect the HSP70 interference plasmid into A549, SW1573 and H460 cells with GenEscort Reagent transfection reagent, add TRAIL to induce cell apoptosis 24 hours later, harvest the treated cells after 18 hours, resuspend in 100 μl sample buffer, and keep aside While oscillating, add 1ml of pre-cooled 70% ethanol dropwise, and fix overnight at 4°C. Then centrifuge at 3000rpm for 5 minutes, suck off the supernatant, add 1ml PI staining solution, mix well, place in the dark at room temperature for more than 30 minutes, and detect within 24 hours. Data were collected and analyzed using CellQuest software.

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 belongs to the field of biological technology, and in particular relates to a method for regulating the expression, quantity and activity of heat shock protein 70 and its application. Utilizing the present invention can also regulate tumor tissue and intracellular transcription factor NF-κB and its activity, death receptor DR4 and DR5 expression and activity, JNK and c-Jun phosphorylation and activity, p53 protein expression and activity, pro-apoptosis Molecule Bax, Bid, t-Bid, capsase 3, capsase 8, capsase 9 expression and pro-apoptotic activity, anti-apoptotic protein c-FLIP and Bcl-2 expression and activity, promote tumor tissue and cell apoptosis. The present invention also relates to the application of the above-mentioned method in the preparation of medicines used in combination with apoptosis-inducing medicines.

Description

1. Technical field: [0001] The invention belongs to the field of biological technology, and in particular relates to a method for regulating the expression, quantity and activity of heat shock protein 70 and its application. 2. Background technology: [0002] According to statistics from the Ministry of Health of China, the total number of new tumor patients in China is about 2.127 million each year, of which, there are about 1.06 million new patients with malignant tumors every year; There are about 1.485 million cancer patients. Taking the morbidity and mortality of lung cancer as an example, about 160,000 people die of lung cancer in the United States every year, and 75% of them are non-small cell lung cancer (NSCLC). In major cities in my country, the incidence of lung cancer has occupied the first place in the incidence of various malignant tumors. Nearly 800,000 people die of lung cancer in my country every year, and non-small cell lung cancer patients account for ab...

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): C12N15/85A61K45/00A61K45/06A61K48/00A61K38/19A61P35/00
Inventor 华子春庄红芹
Owner NANJING UNIV
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