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Serum/plasma miRNA marker for early diagnosis of noncardiac gastric carcinoma and applications thereof

A marker and serum technology, applied in the fields of genetic engineering and oncology

Inactive Publication Date: 2014-07-09
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] However, there are no reports of relatively stable biomarkers for the diagnosis of early non-cardia gastric cancer. If the abnormally expressed serum / plasma miRNAs in early non-cardia gastric cancer can be screened as biomarkers and corresponding diagnostic kits can be developed, It will be a powerful impetus to the diagnosis status of non-cardia gastric cancer in my country, and it will also open up a new way for its drug screening, drug efficacy evaluation and targeted therapy

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  • Serum/plasma miRNA marker for early diagnosis of noncardiac gastric carcinoma and applications thereof
  • Serum/plasma miRNA marker for early diagnosis of noncardiac gastric carcinoma and applications thereof
  • Serum/plasma miRNA marker for early diagnosis of noncardiac gastric carcinoma and applications thereof

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Embodiment 1

[0071] The collection of embodiment 1 sample and the arrangement of sample data

[0072] The inventor has collected a large amount of peripheral blood samples of early stage non-cardia gastric cancer patients, healthy controls and precancerous lesion controls from Jiangsu Provincial People's Hospital and Yangzhou Lidian District since 2006 (the samples used for research are collected in the same period, sampling, Uniform packaging and storage conditions), through the arrangement of the sample data, the inventor selected 230 samples that meet the following standards as experimental samples for TLDA chip detection and subsequent qRT-PCR verification:

[0073] 1. Cases of early (stage I) non-cardia gastric cancer diagnosed by pathology

[0074]2. Patients who have not undergone surgery, radiotherapy and chemotherapy before blood collection

[0075] 3. Healthy controls and precancerous lesions matched with the sex and age of the cases

[0076] The demographic and clinical data o...

Embodiment 2

[0077] TLDA chip detection of miRNAs in embodiment 2 serum / plasma

[0078] 40 patients with early non-cardia gastric cancer and 40 healthy controls in the initial screening stage were detected by TLDA chip to obtain relevant results. The specific steps are:

[0079] 1. Take 600 μl of serum from the early non-cardia gastric cancer case group and the healthy control group, and add 3 times the volume of Trizol reagent;

[0080] 2. Phase separation: place at room temperature for 15 minutes, add final concentration of 10 -4 Pmol / μl of cel-39 (TAKARA) was used as an internal reference, then chloroform was added in an equal volume to the plasma, shaken for 50s, room temperature for 15min, 14,000rpm, 4°C, centrifuged for 15min;

[0081] 3. RNA precipitation: transfer the water phase to a new 15ml centrifuge tube, add 1.5 times the volume of the water phase in absolute ethanol, and mix well;

[0082] 4. Enrich RNA with QIAGEN miRNeasy kit: pipette 700 μl of sample into the spin colu...

Embodiment 3

[0088] qRT-PCR experiment of miRNAs in embodiment 3 serum / plasma

[0089] According to the above TLDA results, miRNAs that meet the following conditions are selected for further verification by qRT-PCR method: 1) C of the two groups of research objects T The values ​​are all less than 30, or only one group of research objects has a CT value of less than 30, and the other group of research objects has a C T The value is less than 35 to improve detection efficiency; 2) ΔΔC T =ΔC T病例 -ΔC T对照 , ΔΔC T Greater than 2, that is, the differential expression is greater than 4 times. Primers for reverse transcription and qRT-PCR were designed for the selected miR-16, miR-25, miR-92a, miR-451 and miR-486-5p (see Table 1). The qRT-PCR detection of miRNAs was performed on individual individuals in the serum of early non-cardia cancer case group and healthy control group. Strict quality control was implemented throughout the study. Each sample was tested three times consecutively. Al...

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Abstract

The invention belongs to the fields of genetic engineering and oncology, and discloses a serum / plasma miRNA marker for early diagnosis of noncardiac gastric carcinoma and applications thereof. The marker is a composition of miR-16, miR-25, miR-92a, miR-451 and miR-486-5p. The marker and primers thereof can be used for preparing a diagnosis kit, and used for assistant early diagnosis of the noncardiac gastric carcinoma.

Description

field of invention [0001] The invention belongs to the fields of genetic engineering and oncology, and relates to a serum / plasma miRNA marker for early diagnosis of non-cardia gastric cancer and application thereof. Background technique [0002] Gastric cancer is one of the most common malignant tumors worldwide. According to the World Health Organization, in 2008 there were about 1 million new cases of gastric cancer worldwide, accounting for 7.8% of all tumors, second only to lung cancer, breast cancer and colorectal cancer. More than 70% of gastric cancer cases occur in developing countries, 50% of which occur in China. In my country, the age-standardized incidence rate of gastric cancer is 41.3 / 100,000 for men and 18.5 / 100,000 for women, second only to lung cancer. About 90%-95% of gastric cancers are adenocarcinomas, originating from the gastric epithelium. Anatomically, it can be divided into cardia cancer and non-cardia cancer. Cardiac carcinoma is an adenocarcino...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 沈洪兵靳光付胡志斌马红霞戴俊程陈佳萍朱陈
Owner NANJING MEDICAL UNIV
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