Application of molecule miR-150 in screening preparation of colorectal cancer detecting medicament

A colorectal cancer and molecular technology, applied in the field of medicinal chemistry, can solve the problem of failing to find miRNA molecules, and achieve great clinical application value and social benefits.

Inactive Publication Date: 2013-10-23
马延磊 +1
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these existing studies only screen miRNAs related to colorectal cancer at the basic level, and use in vitro cell experiments and in vivo animal experiments to confirm their potential biological functions and molec...

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 molecule miR-150 in screening preparation of colorectal cancer detecting medicament
  • Application of molecule miR-150 in screening preparation of colorectal cancer detecting medicament
  • Application of molecule miR-150 in screening preparation of colorectal cancer detecting medicament

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Screening of miR-150 in normal large intestine tissue, adenoma, and adenocarcinoma tissue (Shanghai Sixth People's Hospital) using microfluidic chips and PCR chips

[0042] Extraction of RNA from colorectal normal tissue, adenoma, and adenocarcinoma:

[0043] 1.1 Extraction of total RNA (conventional Trizol method)

[0044] (1) Grind normal large intestine tissue, adenoma, and adenocarcinoma tissue (each 10 mg), add 1 ml TRIzol (Shanghai Invitrogen Company) respectively, and mix well on a vortex shaker.

[0045] (2) Add 1ml of chloroform, mix well by inverting up and down for 1 minute, and let stand at room temperature 25°C for 5 minutes.

[0046] (3) After centrifugation at 12,000 rpm for 15 minutes at 4°C, transfer the supernatant to a new 1.5ml centrifuge tube, add an equal volume of isopropanol, mix by inverting gently, and let stand at room temperature for 5 minutes.

[0047] (4) After centrifugation at 12000 rpm at 4°C for 10 minutes, remove the supern...

Embodiment 2 Embodiment 1

[0067] Example 2 Example 1 miR-150 is verified by PCR

[0068] 1.qRT-PCR detection method:

[0069] 1.3RT (reverse transcription) reaction: operate according to the instructions of TaqMan MicroRNA Reverse Transcription Kit (microRNA reverse transcription kit, Shanghai Ximei Biological Company), add 5 μl of purified total RNA obtained in Example 1 in the reaction (with no ribonuclease The RNA concentration after dilution with water is 0.5-1 μg / μl). The reaction system is as follows: (volume unit: μl)

[0070]

[0071] After adding all the reagents, briefly centrifuge and mix well, obtain cDNA after 30 minutes at 16°C, 30 minutes at 42°C, and 5 minutes at 85°C, and store at -20°C for future use. Then, the reverse transcription product was used as a template, and the ABI7300 quantitative PCR instrument of ABI Company was used to carry out the fluorescent quantitative PCR reaction.

[0072] 1.4Real time PCR reaction: Using the above eDNA as a template and RNU24 as an interna...

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 provides application of molecule miR-150 in screening preparation of a colorectal cancer detecting medicament. The molecule miR-150 is screened out by using a muParaflo microfluidic chip by the inventor of the invention; a paraffin section preparation of a clinical colorectal cancer patient is detected by an established digoxigenin labeled probe of miR-150 and an established detection method of qRT-PCR (Quantitative Real-Time-Polymerase Chain Reaction), respectively; results show that miR-150 low expression level in the clinical colorectal cancer patient indicates short lifetime of the patient and poor sensitivity of the patient to postoperative chemotherapy, and miR-150 high expression level indicates long lifetime of the patient and good sensitivity of the patient to postoperative chemotherapy. Compounds having high sensitivity to miR-150 can be screened by the method, and the compounds can be used for developing a kit for detecting the colorectal cancer. The kit is composed of compounds sensitive to the molecule miR-150 as active ingredients and a medicinal carrier. Simple detection of the miR-150 expression level is advantageous for monitoring of the colorectal cancer patient after healing and determination of chemotherapy sensitivity of the patient, and thus provides an effective method for clinical prevention and treatment of the colorectal cancer; high clinical application value and social benefit are created.

Description

technical field [0001] The invention relates to medicinal chemistry, in particular to the application of biomarker molecule miR-150 in screening, preparing and detecting colorectal cancer drugs. Background technique [0002] Colorectal cancer is one of the common malignant tumors, and its incidence rate ranks third among malignant tumors in the world. With the continuous improvement of living standards and changes in diet structure, the incidence of colorectal cancer in China is also increasing year by year, especially in big cities The growth rate is even faster. For example, during the 20 years from 1984 to 2004, the incidence of colorectal cancer in Shanghai increased from 16 / 100,000 to 23.61 / 100,000, and from 14.26 / 100,000 to 20.43 / 100,000 for women. At present, there are many factors that affect the curative effect of colorectal cancer, and the lack of effective indicators for prognosis and treatment sensitivity monitoring is still an important problem that needs to be ...

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): C12Q1/68
Inventor 马延磊秦环龙张鹏
Owner 马延磊
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