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miRNA (microribonucleic acid) diagnosis and treatment marker for endometrial carcinoma

A technology for endometrial cancer, miRNA-1266, applied in the field of biomedicine

Active Publication Date: 2016-08-31
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few studies on the differential expression of miRNAs in endometrial cancer. Finding and identifying miRNAs related to the occurrence of endometrial cancer provides a basis for clinical diagnosis and treatment of miRNAs, and is helpful for the diagnosis and prognosis of endometrial cancer. Assessment reaches a new level

Method used

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  • miRNA (microribonucleic acid) diagnosis and treatment marker for endometrial carcinoma
  • miRNA (microribonucleic acid) diagnosis and treatment marker for endometrial carcinoma
  • miRNA (microribonucleic acid) diagnosis and treatment marker for endometrial carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1 Screening of miRNAs associated with endometrial cancer

[0066] 1. Sample acquisition: 10 cases of normal endometrial tissue and endometrial cancer tissue were collected respectively. All the above specimens were obtained with the consent of the organizational ethics committee.

[0067] 2. Extraction of total RNA from samples

[0068] Total RNA in advance using QIAGEN Tissue RNA Extraction Kit. Specific steps are as follows:

[0069] 1) In a clean area with less RNase interference, use a mortar containing an appropriate amount of liquid nitrogen to weigh about 20 mg of the tissue sample, and grind it to powder with a pestle;

[0070] 2) Transfer the sample to a 2ml centrifuge tube without RNase;

[0071] 3) Add 300 μl Lysis solution, place in a homogenizer, and grind thoroughly for 1-5 minutes;

[0072] 4) 12000g, 4°C, centrifuge for 10min, transfer the supernatant to a new 1.5ml centrifuge tube;

[0073] 5) Add 600μl RNase-Free Water and mix with a vort...

Embodiment 2

[0096] Example 2 QPCR verification of differentially expressed miRNA-1266

[0097] 1. According to the detection results of the miRNA chip, miRNA-1266 was selected for large-sample QPCR verification. According to the sample collection method in Example 1, 60 endometrial cancer tissue samples and 60 normal endometrial tissue samples were selected.

[0098] 2. The RNA extraction process is the same as in Example 1.

[0099] 3. Reverse transcription:

[0100] 1) Mix 10pg-1μg total RNA template with 2μl 10× buffer, 2μl dATP (10mM), 0.5μl polyA polymerase, 0.5μl ribonuclease (RNase) inhibitor and ribonuclease-free water (RNase freewater), volume The final volume is 20 μl and incubated at 37°C for 1h.

[0101] 2) Add 1 μl of 0.5 μg / μl Oligo(dT) specific RT primer to the reaction tube, and incubate at 70° C. for 5 minutes.

[0102] 3) Immediately incubate on ice for at least 2 minutes to break the secondary structure of RNA and primers.

[0103] 4) Mix the above 20 μl reaction m...

Embodiment 3

[0119] Example 3 Detection of miRNA-1266 expression in endometrial cancer cells by QPCR

[0120] 1. Amplification and identification of miRNA-1266 plasmid

[0121] 1) According to the sequence information of miRNA-1266, Dalian Bao Biotechnology Co., Ltd. designed and synthesized the miRNA-1266 inhibitor plasmid and the negative control plasmid of the random control sequence.

[0122] 2) Plasmid transformation into DH5α competent strain

[0123] ①Take out 100 μl of DH5α competent bacteria from the -80°C refrigerator and melt on ice;

[0124] ② Add 10 μl pMKITeno plasmid to 100 μl DH5α competent bacterial solution, flick and rotate the bottom of the tube to mix well, and place in ice bath for 30 minutes;

[0125] ③ Place in a water bath at 42°C for 90 seconds after heat shock; immediately place in an ice bath for 90 seconds;

[0126] ④ Add 800 μl of Amp-LB culture solution, gently blow and mix, and incubate at 37°C on a constant temperature shaker at 100 rpm for 1 hour to rec...

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Abstract

The invention discloses an miRNA (microribonucleic acid) diagnosis and treatment marker for endometrial carcinoma. The miRNA marker is miRNA-1266. The experiment proves that the miRNA-1266 is related to generation and development of endometrial carcinoma. The in-vitro experiment detects that the inhibition of the miRNA-1266 expression can inhibit the proliferation and transfer of endometrial carcinoma cells and promote the apoptosis of endometrial carcinoma cells, thereby laying a foundation for molecular treatment of endometrial carcinoma.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a miRNA diagnosis and treatment marker for endometrial cancer, specifically the miRNA diagnosis and treatment marker is miRNA-1266. Background technique [0002] Endometrial cancer (EC), also known as uterine body cancer, is a group of epithelial malignant tumors that occur in the endometrium, and adenocarcinoma derived from endometrial glands is the most common. It is one of the three major malignant tumors of the female reproductive tract, accounting for 7% of female systemic malignant tumors and 20%-30% of female reproductive tract malignant tumors. Its mortality rate accounts for 3% of female body tumors, ranking sixth. The incidence of endometrial cancer is closely related to lifestyle, and the incidence rate varies in different regions. In North America and Europe, its incidence rate is second only to breast cancer, lung cancer, and colorectal tumors, ranking first among female rep...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/158C12Q2600/178
Inventor 任静肖武杨承刚
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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