Gastric cancer RNA molecular marker and application thereof
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A marker and molecular technology, applied in the field of biomedicine, can solve problems such as difficulty in finding tumors, inability to effectively screen out gastric cancer patients, lack of sensitivity and specificity, etc.
Active Publication Date: 2016-11-23
INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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[0002] Gastric cancer is one of the most common malignant tumors. Due to the lack of effective early diagnosis methods, it is difficult to detect tumors in early screening. At present, the main diagnostic methods for gastric cancer are gastroscopy, C...
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Embodiment 1
[0021] Gastric cancer tissue RNA detection
[0022] 1. RNA Extraction from Tissue Samples
[0023] Total RNA was extracted with TRI reagent from 49 paired surgical resection specimens of gastric cancer and adjacent tissues, and the integrity of RNA in the tissues was tested by agarose gel electrophoresis.
[0024] 2. RNA reverse transcription
[0025] The reverse transcription reaction was performed with ImProm II Reverse Transcriptase Reaction Kit. Take 1 μg of total tissue RNA as a template for reverse transcription, add 1 μL of random primers, 1 μL of dNTP (10 mM), 4 μL of 5×RT buffer, 0.5 μL of RNase inhibitor, 1 μL of ImProm II reverse transcriptase, and make up to 1 μL with RNase-free water 20 μL. The reaction conditions are: 42°C for 60 minutes, 72°C for 15 minutes, and 20°C for 5 minutes. Select 18S rRNA as the internal reference gene, and take 1 μL of the cDNA obtained by reverse transcription of the tissue and add 9 μL of RNase-free water to dilute it 10 times. ...
Embodiment 2
[0030] Detection of Plasma RNA in Patients with Gastric Cancer
[0031] 1. Plasma RNA Extraction
[0032] Total RNA in 200 μL of plasma was extracted with Qiagen plasma extraction kits from 51 patients with gastric cancer and 53 normal persons matched in gender and age, and dissolved in 40 μL of RNase-free water.
[0033] 2. RNA reverse transcription
[0034] The reverse transcription reaction was performed with ImProm II Reverse Transcriptase Reaction Kit. Take 10 μL of plasma total RNA as a template for reverse transcription, add 1 μL of random primers, 1 μL of dNTP (10 mM), 4 μL of 5×RT buffer, 0.5 μL of RNase inhibitor, 1 μL of ImProm II reverse transcriptase, and make up to 20 μL with RNase-free water . The reaction conditions are: 42°C for 60 minutes, 72°C for 15 minutes, and 20°C for 5 minutes. 18S rRNA was selected as an internal reference gene. The cDNA obtained by reverse transcription of RNA extracted from plasma was used as a template for detecting AK001058, I...
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Abstract
The invention belongs to the field of biological medicine and specifically relates to various RNAs, various combinations of tumor markers thereof and an application thereof in preparing a gastric cancer diagnosis reagent. The invention provides a set of new RNA molecular markers for gastric cancer diagnosis. The content of the set of RNA in gastric cancer tissue is higher than the content of para-carcinoma tissue and the content of the set of RNA in the plasma of gastric cancer patient is higher than the content thereof in the plasma of healthy people. The invention also provides a method for taking one or different combinations of the set of RNA as the tumor markers and the application thereof. The set of RNA molecular marker and the diagnostic method of different combinations of the set of RNA markers adopted for diagnosing gastric cancer are simple in operation, are convenient in material taking and have the characteristics of high specificity, high sensitivity and easiness in mass screening. The set of RNA molecular marker is fit for screening of gastric cancer high-risk groups and auxiliary diagnosis of gastric cancer.
Description
technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to multiple RNAs and tumor markers in different combinations thereof and their application in preparing reagents for diagnosing gastric cancer. Background technique [0002] Gastric cancer is one of the most common malignant tumors. Due to the lack of effective early diagnosis methods, it is difficult to detect tumors in early screening. At present, the main diagnostic methods for gastric cancer are gastroscopy, CT or PET / CT, etc. These inspection methods are costly and invasive. , can not effectively screen out early gastric cancer patients. The classic tumor markers such as CEA, CA724, CA19-9, etc., are limited due to the lack of strong sensitivity and specificity. More and more studies have found that a variety of non-coding RNAs participate in the regulation of tumor cell proliferation, apoptosis and other biological processes, directly or indirectly function as oncogenes...
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IPC IPC(8): C12N15/113C12N15/11C12Q1/68
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 郑晓飞付汉江柯东房学东
Owner INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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