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Gastric cancer related biomarker and application thereof

A technology for gastric cancer and substances, applied in the field of biomedicine, can solve the problem of unclear mechanism of gastric cancer

Active Publication Date: 2018-06-01
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the increasing research on lncRNA, more and more lncRNAs have been found in various tumors. However, the research on lncRNA in gastric cancer is still in its infancy, and the mechanism of action in the occurrence and development of gastric cancer is still unclear, which needs further research

Method used

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  • Gastric cancer related biomarker and application thereof
  • Gastric cancer related biomarker and application thereof
  • Gastric cancer related biomarker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Example 1 Screening of gene markers related to gastric cancer

[0078] 1. Sample collection

[0079] 8 cases of gastric cancer paracancerous tissues and gastric cancer tissue samples were collected, and 3 cases were normal gastric tissues. All cases did not receive chemotherapy and radiotherapy before surgery. All patients had informed consent and obtained the consent of the organizational ethics committee. .

[0080] 2. Preparation of RNA samples

[0081] Tissue RNA was extracted using QIAGEN tissue RNA extraction kit, and the operation was performed according to the specific steps in the manual.

[0082] 3. Total RNA quantification and purity analysis

[0083] Use Bio-Red ultraviolet spectrophotometer to measure the optical density value of total RNA at 280nm and 260nm, when OD 260 / OD 280 When the value of 1.8-2.0, the purity of total RNA is considered reliable and used for the next experiment.

[0084] 4. Microarray analysis of lncRNA expression

[0085] The ...

Embodiment 2

[0090] Example 2 QPCR sequencing to verify the differential expression of the LINC01235 gene

[0091] 1. Large-scale QPCR verification of the differential expression of the LINC01235 gene. According to the sample collection method in Example 1, 50 cases of gastric cancer paracancerous tissues and 50 cases of gastric cancer tissues and 10 cases of normal tissues were selected.

[0092] 2. RNA extraction

[0093] RNA samples were extracted using the QIAGEN Tissue RNA Extraction Kit. For details, please refer to the instruction manual.

[0094] 3. QPCR

[0095] 1) Reaction system:

[0096] Add 1 μl of RNA template, 1 μl of random primers, add double distilled water to 12 μl, mix well, centrifuge at low speed at 65°C for 5 min, then place on ice to cool.

[0097] Continue to add the following components to the 12 μl reaction solution:

[0098] 5× reaction buffer 4μl, RNase inhibitor (20U / μl) 1μl, 10mM dNTP mixture 2μl, AMV reverse transcriptase (200U / μl) 1μl; mix well and cen...

Embodiment 3

[0119] Example 3 Expression of LINC01235 in gastric cancer cell lines

[0120] 1. Cell culture

[0121] Human immortalized gastric mucosal epithelial cell line GES-1, human gastric cancer cell lines HGC-27, MGC-803, and AGS (all purchased from Guangzhou Lydell Biotechnology Co., Ltd.) in the presence of 10% fetal bovine serum and 1% P / S The RPMI1640 culture is based on 37 ℃, 5% CO 2 , Cultivated in an incubator with a relative humidity of 90%. Change the medium once every 2-3 days, use 0.25% EDTA-containing trypsin for routine digestion and passaging, and take the cells in the logarithmic growth phase for the experiment.

[0122] 2. RNA extraction and concentration determination

[0123] Total cellular RNA was extracted using QIAGEN's cellular RNA extraction kit, and the specific operation was referred to the instructions.

[0124] 3. The specific steps of qPCR are the same as in Example 2

[0125] 4. Statistical analysis

[0126] The experiments were repeated three time...

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Abstract

The invention discloses a gastric cancer related biomarker and application thereof. The biomarker is LINC01235. The invention discloses a reagent for detecting the LINC01235 level, a kit, a chip and application thereof in preparation of products for diagnosis of gastric cancer. The invention also discloses a composition containing the LINC01235 inhibitor and application thereof in preparation of drugs for treating gastric cancer.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a biomarker related to gastric cancer and its application. Specifically, the lncRNA is LINC01235. Background technique [0002] Gastric cancer (gastric cancer GC) is one of the malignant tumors of the digestive system that endanger human life and health, and its mortality rate ranks third among tumor-related deaths (Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin, 2011, 61(2):69-90). Gastric cancer is highly common in East Asia, West Asia, and Latin America. The incidence of gastric cancer in China, South Korea, and Japan alone accounts for half of the global total. The morbidity and mortality of gastric cancer in China are both in the forefront of digestive system tumors, accounting for 23.24% of all malignant tumor deaths (ParkinDM, Bray F, Ferlay J, Pisani P. Global cancer statistics, 2002. CA Cancer J Clin, 2005, 55 (2):74-108). A...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886A61K45/00A61P35/00A61P35/04
CPCA61K45/00C12Q1/6886C12Q2600/136C12Q2600/158C12Q2600/178
Inventor 杨承刚宋宏涛李曙光
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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