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

Cyclic RNA (Ribonucleic Acid) biomarker for breast cancer and application of circular RNA biomarker

A breast cancer, annular technology, applied in the field of breast cancer diagnosis, can solve problems such as lack of detection of breast cancer, and achieve the effects of improving the utilization efficiency of medical resources, improving the efficiency of diagnosis, and stabilizing the structure.

Active Publication Date: 2022-05-06
SICHUAN CANCER HOSPITAL
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there is still a lack of specific circRNA biomarkers to efficiently detect breast cancer

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
  • Cyclic RNA (Ribonucleic Acid) biomarker for breast cancer and application of circular RNA biomarker
  • Cyclic RNA (Ribonucleic Acid) biomarker for breast cancer and application of circular RNA biomarker
  • Cyclic RNA (Ribonucleic Acid) biomarker for breast cancer and application of circular RNA biomarker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Preparation method and structure verification of circular RNA circ-GLIS1

[0036] Circ-GLIS1 (hsa_circ_0002079, chr1:54059780-54065951), formed by reverse splicing of the second and third exons of the gene GLIS1 (1p32.3), has a length of 1061 bases. The structural characteristics and sequence information of Circ-GLIS1 are as follows: figure 1 and shown in SEQ No.1.

[0037] Quantitative detection method of circular RNA circ-GLIS1:

[0038] 1. RNA extraction: The total RNA of breast tumor cell MCF-7 was extracted with a total RNA extraction kit, and the purity and concentration of total RNA were detected with a NanoDrop3000 micro-nucleic acid analyzer.

[0039] 2. Reverse transcription: perform reverse transcription to obtain a cDNA template. The reverse transcription reaction steps are: template RNA 500ng, random hexamer primer 1μL, RNase-free water to a total volume of 12μL, mix and store at 70°C Incubate for 5 minutes; then add 4 μL of 5× reaction buffer,...

Embodiment 2

[0043] Example 2 Detection of circular RNA Circ-GLIS1 expression in breast tumor cells and normal breast cells

[0044] Fluorescent quantitative PCR (qPCR) was used to detect the expression of circ-GLIS1 in breast tumor cell MCF-7 and breast normal cell MCF-10A (purchased from the Cell Bank of the Chinese Academy of Sciences, Shanghai), and found that circ-GLIS1 was expressed in breast tumor cells. The expression was significantly higher than that in normal cells, indicating that circ-GLIS1 can act as a tumor-promoting factor and a potential therapeutic target for breast cancer.

[0045] Circular RNA circ-GLIS1 detection method: Total RNA of breast tumor cell MCF-7 and normal breast cell MCF-10A was extracted using a total RNA extraction kit, and the purity and concentration of total RNA were detected with a NanoDrop3000 micro-nucleic acid analyzer. Then, use the aforementioned method to perform reverse transcription to obtain cDNA templates for qPCR detection. The experimenta...

Embodiment 3

[0046] Example 3 Construction of Circ-GLIS1 overexpression vector and its effect on promoting the proliferation and invasion of breast cancer cells

[0047] The pCD5-ciR vector overexpressing circ-GLIS1 was constructed (Guangzhou Jisai Biotechnology Co., Ltd.), the full-length 1061bp sequence of hsa_circ_0002079 was amplified by PCR, and EcoRI and BamHI double enzyme digestion ligated it into pCD5-ciR. The pCD5-ciR vector structure is as follows Figure 4 shown.

[0048] Breast tumor cells were transfected with pCD5-ciR vector overexpressing circ-GLIS1, and the effect of overexpressing circ-GLIS1 on the proliferation and invasion ability of breast tumor cells was observed. Test steps: ① Inoculate MCF-7 cells in a 12-well plate, and transfect when they grow to 70% confluence; ② Add 1 μg circ-GLIS1 pCD5-ciR vector and 2 μL P3000 reagent mixture to 50 μL serum-free medium ③Add 2 μL Lipofectamine 3000 to 50 μL serum-free medium; ④Mix the diluted overexpression vector and Lipofec...

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 discloses a circular RNA biomarker circ-GLIS1 for breast cancer and application of the circular RNA biomarker circ-GLIS1, the structure of the circular RNA circ-GLIS1 is a circular structure formed by connecting a 3'end and a 5 'end in an end-to-end manner and formed by reversely splicing a nucleotide sequence shown in SEQ No.1, and a kit comprises a reagent for quantitatively detecting the biomarker. The invention finds that the expression level of circ-GLIS1 in breast tumor cells is remarkably increased, and proliferation and invasion of breast tumors can be remarkably improved after overexpression of circ-GLIS1; therefore, the circ-GLIS1 can be used as a molecular marker for diagnosing the breast cancer, so that the breast cancer diagnosis is more accurate and quicker, and a new theoretical basis and a new marker are provided for applying the circ-GLIS1 to diagnosis and prognosis analysis of the breast cancer.

Description

technical field [0001] The invention relates to the technical field of breast cancer diagnosis, in particular to the application of a circular RNA circ-GLIS1 biomarker in the preparation of a breast cancer diagnostic reagent. Background technique [0002] Breast cancer is the most common malignancy in women and the leading cause of cancer death in women worldwide. At present, the incidence of breast cancer is increasing year by year, and its comprehensive treatment methods (including surgery, chemotherapy and radiotherapy) have large individual differences and toxic and side effects, which have a great impact on the quality of life of patients. Therefore, finding new breast cancer biomarkers for targeted therapy and individualized therapy has extremely important clinical significance. [0003] Circular RNA (circular RNA, circRNA) is a new type of non-coding RNA with a stable structure and significant tissue and disease specificity. Studies have found that circRNA is involve...

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): C12Q1/6886C12N15/11A61K45/00A61K31/7088A61P35/00A61P15/14
CPCC12Q1/6886A61K45/00A61K31/7088A61P35/00A61P15/14C12Q2600/158C12Q2600/178Y02A50/30
Inventor 王婷龙方懿肖洪涛张晞倩林红
Owner SICHUAN CANCER HOSPITAL
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