Biomarker related to occurrence and development of gastric adenocarcinoma

A technology of gastric cancer and primer pairs, applied in the field of biomarkers, can solve the problem that the functional mechanism of 1ncRNA is not understood by humans.

Inactive Publication Date: 2019-08-13
XUZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although many studies on 1ncRNAs and related databases have emerged in recent years, the functional mechanisms of most 1ncRNAs are still not understood by humans.

Method used

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  • Biomarker related to occurrence and development of gastric adenocarcinoma
  • Biomarker related to occurrence and development of gastric adenocarcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Screening of gene markers associated with gastric cancer

[0047] 1. Sample collection

[0048] The cancer tissues and corresponding paracancerous tissue samples of 4 cases of gastric adenocarcinoma were collected for high-throughput sequencing. All patients did not receive chemotherapy, radiotherapy or endocrine therapy before operation.

[0049] 2. RNA sample preparation and quality analysis

[0050] Total RNA was extracted using Tiangen Animal Tissue Total RNA Extraction Kit (catalogue number: DP431), and the steps are detailed in the instructions.

[0051] 1) Homogenization treatment

[0052] Add 300μl Lysis Solution RL for every 10-20mg tissue, and grind the tissue thoroughly with a grinding pestle; then add 590μl RNase-Free ddH to the homogenate 2 O and 10 μl Proteinase K, mix well and treat at 56°C for 10-20min.

[0053] 2) Centrifuge at 12,000rpm for 2-5min, and take the supernatant for the following operations.

[0054] 3) Slowly add 0.5 times the ...

Embodiment 2

[0086] Example 2 QPCR sequencing verification of differential expression of RP11-320G24.1 gene

[0087] 1. The differential expression of the RP11-320G24.1 gene was verified by large sample QPCR of the cancer tissue samples and paracancerous tissue samples collected from 31 gastric adenocarcinoma patients according to the collection method in Example 1.

[0088] 2. RNA extraction

[0089] The total RNA was extracted using Tiangen Animal Tissue Total RNA Extraction Kit (Cat. No. DP431). See Example 1 for specific steps.

[0090] 3. QPCR

[0091] Primers were designed according to the gene sequences of RP11-320G24.1 and GADPH, the primer sequences are as follows:

[0092] RP11-320G24.1:

[0093] Forward primer: 5′-GTATAATCTGCCACTTCT-3′ (SEQ ID NO.1)

[0094] Reverse primer: 5'-GTCTTCCACATCATTCTA-3' (SEQ ID NO.2)

[0095] GAPDH:

[0096] Forward primer: 5'-AATCCCATCACCATCTTCCAG-3' (SEQ ID NO.3)

[0097] Reverse primer: 5'-GAGCCCCAGCCTTTCTCAT-3' (SEQ ID NO.4)

[0098] The ...

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Abstract

The invention discloses a biomarker related to the occurrence and development of the gastric adenocarcinoma. The expression up-regulation of RP11-320G24.1 in gastric cancer patients is found for the first time through the combination of the high-flux sequencing technology and bioinformatics analysis, it is further verified through QPCR that the RP11-320G24.1 is a differential expression gene, andit is indicated that the RP11-320G24.1 can be applied to the diagnosis and treatment of the gastric cancer as the biomarker.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a biomarker related to the occurrence and development of gastric adenocarcinoma. Background technique [0002] Gastric cancer (GC) is a common malignant tumor in the digestive system with high morbidity and mortality. There are about 1 million new cases of gastric cancer each year worldwide, which seriously threatens human health and quality of life. Although we have made some progress in the clinical treatment and basic research of gastric cancer, many factors are involved in the occurrence and development of gastric cancer, and its pathogenesis is very complicated, and many problems remain to be elucidated. Finding important molecules that effectively regulate the occurrence and development of gastric cancer can guide the development of molecular diagnosis and molecular targeted drugs for clinical gastric cancer, and further research is needed. [0003] Analysis of human genome sequenc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 董东郑骏年宁倩倩李曼
Owner XUZHOU MEDICAL UNIV
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