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Biomarker for diagnosing and treating gastric adenocarcinoma metastasis

A gastric adenocarcinoma and drug technology, which can be used in the field of tumor diagnosis by detecting abnormal KIF14, and can solve problems such as unclear molecular mechanism

Pending Publication Date: 2017-11-24
成都望路医药技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for advanced gastric cancer, the underlying molecular mechanism of invasion and metastasis is still unclear.

Method used

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  • Biomarker for diagnosing and treating gastric adenocarcinoma metastasis
  • Biomarker for diagnosing and treating gastric adenocarcinoma metastasis
  • Biomarker for diagnosing and treating gastric adenocarcinoma metastasis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Differential expression of KIF14 gene

[0064] 1. Sample collection

[0065] 50 cases of metastatic gastric adenocarcinoma tissues and their adjacent normal tissues were collected, and 45 cases of non-metastatic gastric adenocarcinoma tissues and their adjacent normal tissues were collected. The tissue samples were collected with the help of pathologists, and the sample collection standards were as follows: (1) primary gastric adenocarcinoma, no other diseases, and patients who had not undergone cancer radiotherapy and chemotherapy before surgery; (2) samples diagnosed as gastric adenocarcinoma by the pathology department, and the cancer (3) In order to avoid unnecessary cross-contamination, two sets of equipment were used for sampling. First, samples of normal tissue adjacent to cancer that were at least 5 cm away from the cancer tissue without obvious lesions were taken. For full-thickness gastric adenocarcinoma tissue samples, immediately after collection,...

Embodiment 2

[0110] Differential expression of embodiment 2KIF14 protein

[0111] 1. Extract the total protein of the tissue sample in Example 1

[0112] Follow the instructions of the EpiQuik Tissue / Cell Total Protein Extraction Kit for protein extraction.

[0113] 2. Western blot

[0114] β-actin was used as internal reference. 50 μg of total protein was separated by SDS-PAGE, electrotransferred to PVDF membrane, and blocked with 1×TBST containing 5% skimmed milk powder at room temperature for 1 h; added rabbit anti-human KIF14 monoclonal antibody (1:800 dilution) and mouse anti-human β-actin polyclonal antibody (diluted 1:3 000), overnight at 4°C; wash the membrane 4 times with 1×TBST, add goat anti-rabbit and goat anti-mouse IgG (diluted 1:2 000), incubate at room temperature for 1 h; 1×TBST After washing the membrane 4 times, put it in Super Signal chemiluminescent reagent to react for 2 minutes, expose the X-ray film in the dark room, develop and fix it by conventional method.

...

Embodiment 3

[0119] Embodiment 3 interferes with KIF14 gene expression

[0120] 1. Design and synthesis of interference RNA

[0121] According to the KIF14 gene sequence, siRNA was designed and synthesized by Shanghai Gemma Pharmaceutical Technology Co., Ltd. Shanghai Gemma Pharmaceutical Technology Co., Ltd. also provides a negative control siRNA (siRNA-NC) that has no sequence homology with the KIF14 gene.

[0122] siRNA-KIF14:

[0123] The sense strand is 5'-ACAUUCAUUUGAUUUCCAGCC-3' (SEQ ID NO.5);

[0124] The antisense strand is 5'-CUGGAAAUCAAAUGAAUGUCC-3' (SEQ ID NO.6),

[0125] 2. Culture of human gastric adenocarcinoma cells

[0126] Human gastric adenocarcinoma SGC7901 cells were treated with RPMI1640 medium containing 10% fetal bovine serum plus penicillin 100units / ml and streptomycin 100μg / ml, placed at 37°C, 5% CO 2 Cultured in an incubator, the culture medium was changed every 24 hours, and the cells were passaged once every 48 hours. Cells in the logarithmic growth phase w...

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Abstract

The invention discloses that the KIF14 gene can be used as a biomarker for the diagnosis of gastric adenocarcinoma. The experiment of the present invention proves that compared with the normal control tissue, the KIF14 gene expression in the gastric adenocarcinoma tissue is significantly increased, and the interference experiment proves that KIF14 can affect the invasion and migration of the gastric adenocarcinoma. According to the research results of the present invention, drugs capable of inhibiting the expression of the KIF14 gene can be developed, so as to realize the clinical prevention and treatment of gastric adenocarcinoma.

Description

technical field [0001] The present invention relates to the field of tumor diagnosis and treatment, more specifically, the present invention relates to a tumor diagnosis method by means of detecting KIF14 abnormality; and a tumor therapeutic agent for inhibiting KIF14 gene or protein. Background technique [0002] Gastric cancer is a malignant tumor derived from gastric mucosal epithelial cells, mainly adenocarcinoma. In 2008, nearly 1 million cases of gastric cancer were newly diagnosed in the world, and the number of deaths was 740,000. It is the fourth most common cancer in the world and the second most lethal. Although the overall global incidence of gastric cancer has declined, nearly two-thirds of gastric cancer cases are concentrated in economically underdeveloped countries and regions, with high incidence in China and other East Asian countries. According to the latest version of "China Cancer Map" released by "Life Times" in 2014. There are about 3,120,000 new can...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/574A61K45/00A61P35/00
CPCC12Q1/6886A61K45/00C12Q2600/118C12Q2600/158G01N33/57446G01N33/57484
Inventor 宋宏涛马翠
Owner 成都望路医药技术有限公司
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