Gastric cancer tumor marker and application thereof
A tumor marker, gastric cancer technology, applied to gastric cancer tumor markers and their application fields, can solve the problems of low early diagnosis rate of gastric cancer, and achieve simple and accurate diagnosis of gastric cancer, simple, rapid and accurate detection, and is suitable for large-scale promotion and application. Effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0040] Example 1 Quantitative RT-PCR detection of AGR3 gene expression in gastric cancer tissue
[0041] 1. Tissue separation: The experimental tissue was derived from surgical patients with primary gastric cancer. Once the surgically resected stomach was isolated, the lesion and surrounding paracancerous tissues 5 cm away were quickly excised and stored in liquid nitrogen (-80°C). The diagnosis of cancer and paracancer is based on pathological diagnosis.
[0042]2. Extraction of ribonucleic acid (RNA): total RNA was extracted with a Trizol kit (Invitrogen), and the extraction method was referred to the instructions of the Trizol kit. When RNA is ready to be used, the RNA can be precipitated using the following method: add NaAc to 0.3M, centrifuge at 12,000×g for 5 minutes.
[0043] 3. Synthesis of cDNA:
[0044] (1) Add 4 μL of template RNA (0.5 μg / μL), 2 μL of 20 pmol / μL random primer, 5 μL of deionized water into the ice bath centrifuge tube, mix well, and centrifuge for...
Embodiment 2
[0056] Example 2 Quantitative RT-PCR detection of AGR3 gene expression in gastric cancer cell lines
[0057] The expression of AGR3 gene in human gastric epithelial cell line GES-1 and human gastric cancer cell lines BGC823, HGC27, AGS, NCI-N87 and SNU-1 was detected by quantitative RT-PCR. Wherein said cell line comes from Shanghai Cell Bank of Chinese Academy of Sciences.
[0058] According to the method described in Example 1, RNA was extracted from different gastric cancer cell lines, and cDNA was synthesized by reverse transcription. Using GAPDH as an internal control, and using the synthesized cDNA of different gastric cancer cell lines as a template, the PCR reaction was carried out according to the method described in Example 1. Agarose PCR amplification products were detected by gel electrophoresis, and the results were shown in figure 2 .
[0059] Depend on figure 2 The results showed that AGR3 gene was highly expressed in AGS, NCI-N87 and SNU-1 cells, but only ...
Embodiment 3
[0060] Example 3 Expression of AGR3 and changes in cell proliferation ability after knockdown and overexpression of AGR3 in gastric cancer cell lines AGS and NCI-N87
[0061] In vitro chemical synthesis of short hairpin ribonucleic acid (shRNA) has the characteristics of rapidity, simplicity and strong specificity, and has broad application prospects in anti-tumor therapy and other aspects. Using shRNA to study the effect of AGR3 gene on the growth and proliferation of AGS and NCI-N87 cells.
[0062] Two shRNA sequences were designed and synthesized for the AGR3 gene mRNA, respectively AGR3 (s1) and AGR3 (s2), respectively the nucleotide sequence shown in SEQ ID NO:5 and the nucleotide sequence shown in SEQ ID NO:6 ; An irrelevant sequence NC is set as a negative control, such as the nucleotide sequence shown in SEQ ID NO: 9; in addition, the full sequence of the AGR3 gene is chemically synthesized in vitro, as shown in SEQ ID NO: 1; an empty vector (vector) is set as a negati...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com