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Application of micromolecule non-encoding RNA and host gene in diagnosis and treatment of gastric cancer

A non-coding, small molecule technology, applied in the field of oncology, can solve the problem of less research on targeted therapy for gastric cancer

Pending Publication Date: 2021-07-23
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the targeted therapy drug trastuzumab is used clinically to treat patients with Her2-positive gastric cancer, but there are still few studies on targeted therapy for gastric cancer

Method used

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  • Application of micromolecule non-encoding RNA and host gene in diagnosis and treatment of gastric cancer
  • Application of micromolecule non-encoding RNA and host gene in diagnosis and treatment of gastric cancer
  • Application of micromolecule non-encoding RNA and host gene in diagnosis and treatment of gastric cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: Detection of expression levels of microRNA-210 and MIR210HG in clinical samples

[0039] The experiment of the present invention has been approved by the Sixth Affiliated Hospital of Sun Yat-sen University and the Ethics Committee of Nanfang Hospital of Southern Medical University, and the informed consent of the participants has been obtained. The inventors collected 31 pairs and 19 pairs of cancer nests and paracancerous tissues of gastric cancer patients in the Sixth Affiliated Hospital of Sun Yat-sen University and Nanfang Hospital of Southern Medical University, respectively, totaling 50 pairs. After quick-freezing in liquid nitrogen, store at -80°C for later use.

[0040] 1. RNA extraction

[0041] (1) Take 50-100 mg of tissue, preferably add 1 ml of Trizol lysate, shake and mix well, and place in ice bath for 15 min.

[0042] (2) Add 200 μl of pre-cooled chloroform, shake and mix well, and centrifuge at 12000 g for 15 min at 4°C.

[0043] (3) Transf...

Embodiment 2

[0050] Embodiment 2: the making of diagnostic kit

[0051] The kit of the invention is used for gastric cancer detection and gastric cancer diagnosis, and consists of a sample RNA extraction system, a reverse transcription system, a real-time quantitative RT-qPCR system, a primer system and a result analysis system.

[0052] In the sample RNA extraction system of the kit, the inventors used Trizol reagent extraction and isopropanol precipitation to obtain RNA. Preferably, the blood sample is extracted with Trizol reagent added with external reference NC67, and further combined with phenol / chloroform extraction and purification, and glycogen-assisted precipitation to obtain RNA. The inventors used TAKARA's PrimeScript RT reagent Kit with gDNAEraser reverse transcription kit for reverse transcription. The inventors used Ribo primers or self-designed primers as the primer system of the kit, including primers for microRNA-210, MIR210HG, U6, ACTB and NC67 (external reference). In...

Embodiment 3

[0053] Example 3: In vitro transfer chamber experiment

[0054] The in vitro metastasis chamber experiment is an experiment to evaluate the migration and invasion ability of tumor cells by detecting and counting the tumor cells that migrate to the other side of the basement membrane of the metastasis chamber. The present invention studies the effects of microRNA-210 and MIR210HG on the migration ability of gastric cancer cells in vitro through the following in vitro transfer chamber experiments combined with loss of function experiments and acquired function experiments:

[0055] 1. Loss of sexual function experiment

[0056] (1) Using siRNA or microRNA antisense nucleotides to knock down the gene or microRNA respectively. The above siRNAs are siRNAs of MIR210HG: siMIR210HG-1 and siMIR210HG-2 are siRNAs targeting different sites of MIR210HG respectively, and the negative control is NC; the microRNA antisense nucleotide is the antisense nucleotide Anti-miR of microRNA-210 -21...

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Abstract

The invention discloses a micromolecule non-encoding RNA (microRNA-210), a host gene (lncRNA MIR210HG) of the micromolecule non-coding RNA (microRNA-210) and application thereof. The microRNA-210 and the lncRNA MIR210HG disclosed by the invention can be used as specific markers of the gastric cancer. Besides, an in-vitro metastasis chamber and an in-vivo mouse tail intravenous injection tumor cell lung metastasis model are carried out, which shows that the microRNA-210 and the lncRNA MIR210HG can promote gastric cancer cell metastasis, and the gastric cancer cell metastasis can be inhibited by inhibiting a common transcription factor of the microRNA-210 and the lncRNA MIR210HG. Therefore, the microRNA-210, the lncRNA MIR210HG and the common transcription factor of the microRNA-210 and the lncRNA MIR210HG can be used for gastric cancer diagnosis, can also be used as targets for preparing drugs for inhibiting gastric cancer metastasis or anti-tumors, and can provide a new thought and a new way for gastric cancer treatment.

Description

technical field [0001] The present invention relates to a small molecule non-coding RNA (microRNA-210) and its host gene (lncRNAMIR210HG), in particular to the application of the small molecule non-coding RNA (microRNA-210) and its host gene (lncRNA MIR210HG) in the diagnosis and treatment of gastric cancer. in the field of oncology. Background technique [0002] Gastric cancer (Gastric Cancer) is the fourth most common cancer in the world, ranking fourth among men and fifth among women worldwide. Although the incidence of gastric cancer has decreased in some countries, due to its high lethality, gastric cancer ranks second in the world in cancer-related deaths. my country is a country with a high incidence of gastric cancer. The latest statistics show that in China, the incidence of gastric cancer is as high as 679.1 per 100,000 people, and the death rate is as high as 498.0 per 100,000 people. In recent years, thanks to the popularization of gastrointestinal endoscopy an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12Q1/6886
CPCC12N15/113C12Q1/6886C12N2310/141C12N2310/11C12Q2600/158C12Q2600/178C12Q2600/136
Inventor 林雪嘉李志勇邓明霞
Owner JINAN UNIVERSITY
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