Long non-coding RNA (lncRNA) marker derived from adipocyte exosomes and application and product of lncRNA marker

A technology of adipocytes and exosomes, applied in the field of biomedicine, can solve problems such as exosomes without TUG1 gene, and achieve the effect of improving prognosis

Inactive Publication Date: 2020-07-28
余之刚
View PDF3 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, most of the TUG1 genes studied in the prior art are derived from tumor cells and act on tumor cells. There is no related research on TUG1 gene exosomes. The present invention studies breast cancer itself and tumor stromal cells. The expression of lncRNA in the body, as well as the regulation of the lncRNA on the biological behavior of breast cancer, provides a lncRNA marker derived from adipocyte exosomes and its application and products, which are helpful for the accurate diagnosis and personalized diagnosis of breast cancer Chemotherapy is of great importance

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
  • Long non-coding RNA (lncRNA) marker derived from adipocyte exosomes and application and product of lncRNA marker
  • Long non-coding RNA (lncRNA) marker derived from adipocyte exosomes and application and product of lncRNA marker
  • Long non-coding RNA (lncRNA) marker derived from adipocyte exosomes and application and product of lncRNA marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Induce adipocyte formation in vitro, and use PA to stimulate lipid droplet hypertrophy, and simulate the process of adipocyte hypertrophy in vitro.

[0074] 1. Induction and differentiation of 3T3-L1 preadipocyte cell line

[0075] 1) Inoculate 3T3-L1 preadipocytes on a culture plate, and use high-glucose DMEM medium containing 10% fetal bovine serum at 37°C, 5% CO 2 to cultivate.

[0076] 2) After the cells reach 80-90% confluence and confluence for 2 days (fusion means to let the cells contact and inhibit, that is, change the medium after confluence to allow them to grow for another two days), add the final concentration of 0.5mmol / L IBMX (stock solution concentration 0.5mmol / L), dexamethasone at a final concentration of 1umol / L (stock solution concentration of 1mg / ml (2.5mmol / L)) and insulin at a final concentration of 10ug / ml in 10% fetal bovine serum Incubate in high-sugar DMEM for 48 hours; (the inducer is then added to the culture medium and mixed evenly before...

Embodiment 2

[0096] After exosomes were extracted from the supernatant of adipocytes, they were identified under the electron microscope.

[0097] 1. Supernatant Collection

[0098] 1) 3T3-L1 preadipocytes were induced with PA for 24 hours after adipogenesis, and BSA was used as a control.

[0099] 2) After 24 hours, replace the culture medium with serum-free DMEM medium.

[0100] 3) After 24 hours, the supernatant was collected and stored in a -80 refrigerator for later use.

[0101] 2. Exosome Extraction

[0102] 1) The supernatant after induction with equal volumes of PA and BSA was taken out from the -80 refrigerator and thawed on ice.

[0103] 2) After the supernatant is filtered through a 0.22um pore size filter, it is concentrated using a 100K ultrafiltration tube.

[0104] 3) Add ExoQuick-TC Separation Reagent to the concentrated supernatant at a ratio of 1:5, mix by inverting, and let stand overnight in a refrigerator at 4°C.

[0105] 4) Take out the concentrated supernatant ...

Embodiment 3

[0121] After extracting exosomes from adipocyte supernatant, nanoparticle tracking analysis and identification.

[0122] Exosome extraction method is the same as in Example 2

[0123] Nanoparticle Tracking Analysis (NTA) is to track and analyze the Brownian motion of each particle, and calculate the hydrodynamic diameter and concentration of nanoparticles in combination with the Stockes-Einstein equation. NTA technology has been recognized by the exosome research field as one of the exosome characterization methods; this part is assisted by the School of Chemistry, Shandong University.

[0124] Experimental results:

[0125] As attached in the manual Figure 5 and Figure 6 As shown, in the supernatant induced by PA and BSA, after exosomes were collected, nanoparticle size analysis showed that the peak particle size distribution was within 200 nm, which was consistent with the physiological characteristics of exosomes.

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

PropertyMeasurementUnit
diameteraaaaaaaaaa
particle sizeaaaaaaaaaa
pore sizeaaaaaaaaaa
Login to view more

Abstract

The invention discloses a long non-coding RNA (lncRNA) marker derived from adipocyte exosomes and application and product of the lncRNA marker. The lncRNA marker is a long non-coding RNA TUG1 gene orhomolog, mutant or isotype thereof; the lncRNA TUG1 is derived from exosomes secreted by breast cancer cells, tumor interstitial adipocytes or tumor interstitial fibroblasts, or body fluid exosomes; and the lncRNA TUG1 is highly expressed in mast adipocytes and breast cancer, and can be delivered to breast cancer cells from the adipocytes through the exosomes. The lncRNA marker derived from the adipocyte exosomes is a product for detecting the long non-coding RNA TUG1 of the tumor interstitial adipocytes and the long non-coding RNA TUG1 of tumor, and lays the foundation for early diagnosis ofthe breast cancer; and as a therapeutic target, and as an indicator of disease treatment, the long non-coding RNA TUG1 is applied clinically, and thus, a theoretical basis is provided for prevention and mechanism research of the breast cancer.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to an lncRNA marker derived from adipocyte exosomes and its application and product. Background technique [0002] Breast cancer is one of the most common malignancies in women worldwide. In recent years, the morbidity and mortality of breast cancer in Chinese women have been on the rise. At present, the incidence rate ranks first among female malignant tumors in my country, and the age of onset has tended to be younger in recent years. At present, the treatment methods for breast cancer mainly include: surgical treatment, (neo)adjuvant chemotherapy, adjuvant radiotherapy, endocrine therapy and targeted therapy, etc. Reasonable treatment methods at different stages can obtain better therapeutic effects. However, there are still some patients with recurrence and distant metastasis, which are the main cause of death of breast cancer patients, seriously affecting the physical and ...

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 Applications(China)
IPC IPC(8): C12Q1/6886A61K45/00A61K31/7088A61P35/00A61P15/14
CPCA61K31/7088A61K45/00A61P15/14A61P35/00C12Q1/6886C12Q2600/118C12Q2600/136C12Q2600/158C12Q2600/178
Inventor 余之刚黄淑亚周文重刘丽媛王斐郭明明
Owner 余之刚
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap