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

Application of reagent for detecting ring-shaped RNA (ribonucleic acid) circRNF13 in preparing tongue squamous carcinoma auxiliary diagnosis preparation

A technology for auxiliary diagnosis and detection reagents, applied in the field of tumor molecular biology, can solve the problems of biological function and mechanism of action that have not been reported in any way

Active Publication Date: 2018-07-20
CENT SOUTH UNIV
View PDF3 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

We found that the expression of circRNF13 changed most significantly, suggesting that circRNF13 may be related to cisplatin resistance in tongue squamous cell carcinoma, but so far, the biological function and mechanism of circRNF13 have not been reported

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
  • Application of reagent for detecting ring-shaped RNA (ribonucleic acid) circRNF13 in preparing tongue squamous carcinoma auxiliary diagnosis preparation
  • Application of reagent for detecting ring-shaped RNA (ribonucleic acid) circRNF13 in preparing tongue squamous carcinoma auxiliary diagnosis preparation
  • Application of reagent for detecting ring-shaped RNA (ribonucleic acid) circRNF13 in preparing tongue squamous carcinoma auxiliary diagnosis preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1, PCR sequencing determined the full-length sequence of circRNF13 in tongue squamous cell carcinoma cells

[0038] 1. Materials and methods

[0039] 1.1 Cell lines

[0040] Tongue squamous cell carcinoma cells Tca8113 and Cal27 were purchased from the Cell Center of Central South University and cultured under conventional conditions.

[0041] 1.2 Reagents and kits

[0042] TRIZOL TM Reagent (Invitrogen); Gel Recovery Kit (OMEGA); Reverse Transcription Kit (Promega); Proteinase K, DNase I, RNAsin, RNase A (GBICOL Company); Enzyme (Takara).

[0043] 1.3 Real-time quantitative PCR detection of circRNF13 expression in tongue squamous cell carcinoma cells

[0044] Total RNA was extracted, and 1 μg of RNA was reverse-transcribed into cDNA, followed by real-time fluorescent quantitative PCR. The forward primer of circRNF13 is 5-GTCCAGGATAGACATAGAGC-3, as shown in SEQ NO:2, and the reverse primer is 5-GTGTAGACTTGTGTGGCTGA-3. As shown in SEQ NO:3.

[0045] The...

Embodiment 2

[0068] Example 2, circRNF13 inhibits cell cycle arrest and induces apoptosis in tongue squamous cell carcinoma cells

[0069] 1. Materials and methods

[0070] 1.1 Reagents and kits

[0071] Restriction enzymes Cla I and Sac II and T4DNA ligase were purchased from TakaRa Company; TRIZOL TM Reagent (Invitrogen); Plasmid Extraction Kit, Gel Recovery Kit (OMEGA); Reverse Transcription Kit (Promega); Proteinase K, DNase I, RNAsin, RNase A (GBICOL Company); Tetramethylazolazolium Blue ( MTT, Sigma); antibiotic G418 (Ameresc); cell cycle detection kit (Invitrogen), apoptosis detection kit (Invitrogene).

[0072] 1.2 Induction and culture of tongue squamous cell carcinoma drug-resistant cells

[0073] In the cell culture medium, a low dose of cisplatin was added, and the concentration of cisplatin was gradually increased. After a long period of induction culture, a drug-resistant cell line resistant to cisplatin was finally obtained.

[0074] 1.3 Construction of circRNF13 eukaryo...

Embodiment 3

[0151] Example 3, real-time fluorescent quantitative PCR method detection confirmed the down-regulation of circRNF13 in tongue squamous cell carcinoma

[0152] 1. Materials and methods:

[0153] 28 adjacent tongue squamous cell carcinoma tissues and 28 tongue squamous cell carcinoma tissues were collected, total RNA was extracted, 1 μg RNA was reverse transcribed into cDNA, and real-time fluorescent quantitative PCR was performed. The forward primer of circRNF13 is 5-GTCCAGGATAGACATAGAGC-3, as shown in SEQ NO:2, and the reverse primer is 5-GTGTAGACTTGTGTGGCTGA-3. As shown in SEQ NO:3.

[0154] The GAPDH forward primer used for the control is 5'-ACCACAGTCCATGCCATCAC-3' as shown in SEQ NO:4, and the reverse primer 5'-TCCACCACCCTGTTGCTGTA-3' as shown in SEQ NO:5.

[0155] Real-time fluorescence quantitative PCR reaction system

[0156]

[0157] Real-time fluorescent quantitative PCR reaction steps

[0158]

[0159]

[0160] After the reaction, the amplification curve...

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
Particle sizeaaaaaaaaaa
Login to View More

Abstract

The invention discloses application of a reagent for detecting ring-shaped RNA (ribonucleic acid) circRNF13 in preparing a tongue squamous carcinoma auxiliary diagnosis preparation, and particularly relates to a preparation for auxiliary diagnosis of tongue squamous carcinoma through a real-time fluorescence quantification analysis method. The research proves that the expression of circRNF13 in tongue squamous carcinoma tissue is down-regulated. Therefore, the expression situation of the circRNF13 is used for the auxiliary diagnosis of the tongue squamous carcinoma and has far-reaching clinical significance and important popularization and application prospects.

Description

technical field [0001] The invention belongs to the field of tumor molecular biology, and in particular relates to the application of a reagent for detecting circular RNA circRNF13 in the preparation of auxiliary diagnostic preparations for tongue squamous cell carcinoma. Background technique [0002] Human Genome Project and its follow-up DNA Element Encyclopedia Project (The Encyclopedia of DNAElements Project, ENCODE) research results show that protein-coding gene sequences only account for 1-3% of the human genome sequence, while most of the human genome can be transcribed The sequence is non-coding RNA (non-coding RNA, ncRNA). Although non-coding RNA does not encode protein, it has been attracting much attention in the field of biomedical research due to its extensive involvement in the regulation of gene expression in cells. The most studied are linear ncRNA molecules, which can be divided into microRNA (microRNA, miRNA, 19-23nt) and long non-coding RNA (long non-codi...

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
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/118C12Q2600/166C12Q2600/178
Inventor 刘凌云郭灿熊芳曾朝阳熊炜张姗姗龚朝建王裕民莫勇真李夏雨李小玲李桂源
Owner CENT SOUTH 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