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

Application of miR-27a and diagnostic kit thereof

A mir-27a, 1.mir-27a technology, applied in the fields of biotechnology and medicine, can solve the problems of MET and EGFR expression reduction, and achieve great application value and great practical significance

Active Publication Date: 2014-08-06
JINING MEDICAL UNIV
View PDF4 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In non-small cell lung cancer, miR-27a targets the 3'UTR of MET and EGFR leading to decreased expression of MET and EGFR

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 miR-27a and diagnostic kit thereof
  • Application of miR-27a and diagnostic kit thereof
  • Application of miR-27a and diagnostic kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Differential expression of miR-27a in colon cancer tissue samples

[0040] 1. Collection of human colon tissue specimens

[0041] From November 2012 to December 2013, a total of 41 colon cancer specimens and their adjacent normal colon tissue specimens were collected from Xinxiang Central Hospital and the First Affiliated Hospital of Xinxiang Medical College. A part was stored in liquid nitrogen, -80 ℃ refrigerator, and RNA and protein were extracted for real-time quantitative PCR and western blotting. The other part was fixed with 10% formalin and then embedded in paraffin. The clinical and pathological information of the patients is shown in Table 1. All patients informed in writing that specimen collection was approved by the Ethics Review Committee of Xinxiang Medical College.

[0042] Table 1 Association between miR-27a and clinicopathological data

[0043]

[0044] N: represents the number of patients; n: represents the number of patients in each ...

Embodiment 2

[0070] Example 2miR-27a inhibits distant metastasis of colon cancer and migration of colon cancer cells

[0071] 1. Cell culture

[0072] Human colon cancer cell lines HCT116, Caco2 and SW480 were cultured in complete MEM medium. Human normal colon epithelial cell line NCM460 was purchased from Shanghai Bioleaf (Shanghai, China) and cultured with RPMI-1640. The mouse colon adenocarcinoma cell line MC38 (from C57BL6 / J mice) was cultured in MEM (Invitrogen, Carlsbad, CA). All media contained 10% fetal bovine serum and antibiotics (10000 U / ml penicillin, 10 ug / ml streptomycin). The cell incubator environment is 37°C, humidified, 5% CO 2 .

[0073] 2. Scratch test

[0074] HCT116 cells were cultured in a 100mm petri dish, and HCT116 cells were transiently transfected with miR-27a and negative control miRNA. A 10 μl pipette tip was used to make a scratch on the bottom of the petri dish, and the culture was continued, and observed at 24 and 48 h, respectively.

[0075] see f...

Embodiment 3

[0076] Example 3 miR-27a targets smad2 and SGPP1

[0077] 1. Cell culture

[0078] Human colon cancer cell lines HCT116, Caco2 and SW480 were cultured in complete MEM medium. Human normal colon epithelial cell line NCM460 was purchased from Shanghai Bioleaf (Shanghai, China) and cultured with RPMI-1640. The mouse colon adenocarcinoma cell line MC38 (from C57BL6 / J mice) was cultured in MEM (Invitrogen, Carlsbad, CA). All media contained 10% fetal bovine serum and antibiotics (10000 U / ml penicillin, 10 μg / ml streptomycin). The cell incubator environment is 37°C, humidified, 5% CO 2 .

[0079] 2. Immunohistochemical staining

[0080] (1) Making slices. A tissue wax block was taken, a piece was cut and placed on a glass slide, and then placed in a 60°C oven for 60 min.

[0081] (2) Dewaxing. The slices were taken out from the oven and placed in xylene I and II for 15 min each, 100% ethanol I and II for 10 min each, 95% ethanol I and II for 10 min each, 80% ethanol for 10 m...

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 fields of biotechnology and medical technology, provides application of miR-27a in preparation of a kit for diagnosing colon cancer or colon cancer metastasis, further provides application of miR-27a in preparation of a drug for inhibiting the colon cancer or colon cancer metastasis, and further provides a diagnostic kit for colon cancer or colon cancer metastasis. The diagnostic kit comprises real-time fluorescent quantitative PCR (Polymerase Chain Reaction) primers containing miR-27a, wherein the primers respectively are as follows: the upstream primer sequence is 5'-TTCGGTTCACAGTGGCTAAG-3'; the general downstream primer sequence is 5'-CAGTGCAGGGTCCGAGGT-3'. The kit provided by the invention can perform early diagnosis on the colon cancer by detecting expression of miR-27a, and evaluate prognosis for a patient by detecting the expression level and the relation between clinical stages and metastasis.

Description

technical field [0001] The invention belongs to the fields of biotechnology and medical technology, and particularly relates to the application of miR-27a to the diagnosis of colon cancer occurrence or metastasis. Background technique [0002] Colon cancer is one of the most common malignant tumors in the world, and the reasons for the occurrence and development of colon cancer are not fully understood. Lymph node metastasis and distant metastasis of colon cancer are the main causes of death of colon cancer patients. The prognosis of patients with colon cancer distant metastasis is extremely poor. Therefore, the research on the occurrence and metastasis of colon cancer has always been the basic and clinical research. hot spot. The conventional treatments for colon cancer are surgery, radiotherapy and chemotherapy. The rapid development of immunotherapy and gene therapy in recent years has shown great application potential. Recent studies have shown that epigenetic changes...

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): C12Q1/68A61K48/00A61P35/00
CPCA61K31/7088C12Q1/6886C12Q2600/112C12Q2600/118C12Q2600/158C12Q2600/178
Inventor 鲍永华杨万才郭永臣李泽信薛会朝朱绍辉徐红伟游焜陈志国赵铁索李凯王倩
Owner JINING MEDICAL 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