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Application of linc01389 in the diagnosis and treatment of gastric cancer

A gastric cancer and reagent technology, applied in the field of biomedicine, can solve the problems of lack of high diagnostic sensitivity and specificity, and the inability of early detection or screening for gastric cancer patients.

Active Publication Date: 2020-09-01
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Gastroscopy and PET / CT are the most commonly used methods for clinical diagnosis of gastric cancer, but early detection or screening of gastric cancer patients cannot be achieved
Although some markers such as CEA, CA-199, CA-724, etc. have been applied to the diagnosis of gastric cancer, these tumor markers do not have high diagnostic sensitivity and specificity, and their clinical application has great limitations.

Method used

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  • Application of linc01389 in the diagnosis and treatment of gastric cancer
  • Application of linc01389 in the diagnosis and treatment of gastric cancer
  • Application of linc01389 in the diagnosis and treatment of gastric cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Example 1 Screening of gene markers associated with gastric cancer

[0087] 1. Sample collection

[0088] 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. .

[0089] 2. Preparation of RNA samples

[0090] Tissue RNA was extracted using QIAGEN tissue RNA extraction kit, and the operation was performed according to the specific steps in the manual.

[0091] 3. Total RNA quantification and purity analysis

[0092] 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.

[0093] 4. Microarray analysis of lncRNA expression

[0094]...

Embodiment 2

[0099] Example 2 QPCR sequencing to verify the differential expression of LINC01389 gene

[0100] 1. Large-scale QPCR verification of the differential expression of the LINC01389 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 gastric tissues were selected.

[0101] 2. RNA extraction

[0102]RNA samples were extracted using the QIAGEN Tissue RNA Extraction Kit. For details, please refer to the instruction manual.

[0103] 3. QPCR

[0104] 1) Reaction system:

[0105] 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.

[0106] Continue to add the following components to the 12 μl reaction solution:

[0107] 5× reaction buffer 4μl, RNase inhibitor (20U / μl) 1μl, 10mM dNTP mixture 2μl, AMV reverse transcriptase (200U / μl) 1μl; mix well and ...

Embodiment 3

[0128] Example 3 Expression of LINC01389 in gastric cancer cell lines

[0129] 1. Cell culture

[0130] 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.

[0131] 2. RNA extraction and concentration determination

[0132] Total cellular RNA was extracted using QIAGEN's cellular RNA extraction kit, and the specific operation was referred to the instructions.

[0133] 3. The specific steps of qPCR are the same as in Example 2

[0134] 4. Statistical analysis

[0135] The experiments were repeated three time...

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Abstract

The invention discloses application of LINC01389 in diagnosis and treatment of stomach cancer. The application is characterized in that an lncRNA expression spectrum chip technique is utilized, a bioinformatics technique is adopted to analyze, and the differently-expressed lncRNA LINC01389 in stomach cancer tissues is screened; verified by a large sample, the up-regulated expression of LINC01389 in the stomach cancer tissues is further proofed; proofed by cell experiment, the interference to the expression level of LINC01389 can influence the proliferation, transfer and invasion of stomach cancer cells, so as to provide a molecular target for the treatment of stomach cancer.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of LINC01389 in the diagnosis and treatment of gastric cancer. Background technique [0002] Gastric cancer is one of the most common malignant tumors in the world. Its incidence rate ranks fourth among many tumors, and its mortality rate ranks second among tumor-related deaths (SIEGEL R L, MILLER K D, JEMAL A. Cancer statistics, 2016[ J]. CA: a cancer journal for clinicians, 2016, 66(1): 7-30). Due to the lack of effective early screening methods, when gastric cancer is detected in patients, the tumor is mostly in advanced stage or has already occurred distant metastasis (WANG X N, LIANGH. Some problems in the surgical treatment of gastric cancer[J]. Chinese journal of cancer, 2010,29(4):369-73), which leads to the poor prognosis of patients with gastric cancer after treatment and surgery. Up to now, the molecular mechanisms related to the occurrence, development and met...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/136C12Q2600/158C12Q2600/178
Inventor 石小峰任静马翠
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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