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Application of MCM8 as gastric adenocarcinoma metastasis marker

A gastric adenocarcinoma and drug technology, which is applied in the field of tumor diagnosis with the detection of MCM8 abnormality as a means, can solve the problems of unclear molecular mechanism and the like

Active Publication Date: 2017-11-24
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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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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  • Application of MCM8 as gastric adenocarcinoma metastasis marker
  • Application of MCM8 as gastric adenocarcinoma metastasis marker
  • Application of MCM8 as gastric adenocarcinoma metastasis marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Differential expression of MCM8 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

[0111] Example 2 Differential expression of MCM8 protein

[0112] 1. Extract the total protein of the tissue sample in Example 1

[0113] Follow the instructions of the EpiQuik Tissue / Cell Total Protein Extraction Kit for protein extraction.

[0114] 2. Western blot

[0115] 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 MCM8 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.

[011...

Embodiment 3

[0120] Example 3 Interference with MCM8 gene expression

[0121] 1. Design and synthesis of interference RNA

[0122] According to the MCM8 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 MCM8 gene.

[0123] siRNA-MCM8:

[0124] The sense strand is 5'-CACAGTTTTTTGCTTTCAACAAAG-3' (SEQ ID NO.5);

[0125] The antisense strand is 5'-TGGATCGATTCATACCATATAAA-3' (SEQ ID NO.6),

[0126] 2. Culture of human gastric adenocarcinoma cells

[0127] 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 MCM8 serving as a diagnosis and treatment target of gastric adenocarcinoma. Therefore, the MCM8 can be used for developing products for diagnosing the gastric adenocarcinoma and developing medicines for treating the gastric adenocarcinoma. The research achievement of the invention provides theoretical basis for a clinicist to formulate personalized treatment programs and can provide a new medicine target point for developing gastric adenocarcinoma medicines.

Description

technical field [0001] The invention relates to the field of tumor diagnosis and treatment, more specifically, the invention relates to a tumor diagnosis method by means of detecting MCM8 abnormality; and a tumor therapeutic agent for inhibiting MCM8 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 cancer cases in my co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/574A61K45/00A61P35/00
CPCA61K45/00G01N33/57446G01N33/57484C12Q1/6886C12Q2600/158C12Q2600/118
Inventor 宋宏涛马翠
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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