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Application of CHKA gene to preparation of esophageal cancer diagnosis and treatment product

A technology for esophageal cancer and products, applied in the biological field, can solve the problems of high degree of malignancy, difficulty in early diagnosis of esophageal cancer, poor treatment effect and prognosis of esophageal cancer

Active Publication Date: 2016-08-24
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, early diagnosis of esophageal cancer is difficult, and the degree of malignancy is high. Once diagnosed, it has reached an advanced stage. Therefore, the current treatment effect and prognosis of esophageal cancer are still poor, and the 5-year survival rate is still lower than 30%.

Method used

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  • Application of CHKA gene to preparation of esophageal cancer diagnosis and treatment product
  • Application of CHKA gene to preparation of esophageal cancer diagnosis and treatment product
  • Application of CHKA gene to preparation of esophageal cancer diagnosis and treatment product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Differential expression of embodiment 1CHKA gene

[0061] 1. Experimental materials:

[0062] Esophageal cancer tissue is the specimen removed by thoracic surgery in the hospital, and normal esophageal tissue is the specimen taken out under the gastroscope. surgical blade cut.

[0063] All cases were confirmed pathologically. Including 40 cases of esophageal cancer tissue and 30 cases of normal esophageal tissue. Among them, there were 34 cases of esophageal squamous cell carcinoma and 6 cases of esophageal adenocarcinoma. Normal esophageal tissue was used as the control group. All patients were pathologically confirmed before operation, and no radiotherapy, chemotherapy and other treatments were performed before operation, and no tumors in other parts were found. All tissues were stored in liquid nitrogen within half an hour of the operation, and then transferred to a -80°C refrigerator for freezing.

[0064] 2. Acquisition of RNA from esophageal cancer tissue an...

Embodiment 2

[0074] Differential expression of embodiment 2CHKA protein

[0075] 1. The research object is the same as in Example 1.

[0076] 2. Extract tissue total protein

[0077] Follow the instructions of the EpiQuik Tissue / Cell Total Protein Extraction Kit for protein extraction.

[0078] 3. Western blot detection

[0079] The extracted protein was quantitatively subjected to SDS-PAGE electrophoresis, followed by membrane transfer, blocking, primary antibody incubation, secondary antibody incubation, and color development.

[0080] 4. Statistical processing

[0081] The gray value of the protein band was analyzed using Image J software, and the gray value of the CHKA protein band was normalized with β-actin as an internal reference. The results and data are expressed in the form of mean ± standard deviation, and SPSS13.0 statistical software is used for statistical analysis. The difference between the two is tested by t test, and it is considered statistically significant when P<...

Embodiment 3

[0084] Embodiment 3 inhibits CHKA gene expression

[0085] 1. siRNA design and synthesis

[0086] siRNA sequences against CHKA:

[0087] siRNA1-CHKA:

[0088] The sense strand is 5'-AUUCUUCAGUAUCUAAUCGCC-3' (SEQ ID NO.7);

[0089] The antisense strand is 5'-CGAUUAGAUACUGAAGAAUUA-3' (SEQ ID NO.8),

[0090] siRNA2-CHKA:

[0091] The sense strand is 5'-AUGAAAUGUAGCCAUUUUCUC-3' (SEQ ID NO.9);

[0092] The antisense strand is 5'-GAAAAUGGCUACAUUUCAUGG-3' (SEQ ID NO.10),

[0093] siRNA3-CHKA:

[0094] The sense strand is 5'-AAGAUAUUACCUUCUUGACAG-3' (SEQ ID NO.11);

[0095] The antisense strand is 5'-GUCAAGAAGGUAAUAUCUUGU-3' (SEQ ID NO.12)

[0096] The above siRNA sequences and negative control siRNA sequences (siRNA-NC) were provided by Shanghai Gemma Pharmaceutical Technology Co., Ltd.:

[0097] 2. Culture and transfection of esophageal cancer cells

[0098] 2.1 Cell culture

[0099] The esophageal cancer cell line ECA109 was inoculated in DMEM medium containing 10% fetal...

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Abstract

The invention discloses a CHKA gene that can function as a molecular marker for early diagnosis of esophageal cancer. Experiments prove that compared with normal esophageal tissues, the CHKA gene in esophageal cancer tissues has significantly increased expression, and RNA interference experiments prove that CHKA can affect proliferation of esophageal cancer cells. According to study results herein, it is possible to develop medicine to inhibit CHKA gene expression so as to clinically prevent and treat the esophageal cancer.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of CHKA gene in the diagnosis and treatment of esophageal cancer. Background technique [0002] Esophageal cancer, as a malignant tumor of the digestive system, seriously threatens people's health and affects their quality of life. According to epidemiological surveys, my country is a high-incidence area of ​​esophageal cancer in the world, with more than 150,000 deaths due to esophageal cancer every year, ranking first in the number of deaths from esophageal cancer in the world. The occurrence of esophageal cancer is closely related to dietary habits, carcinogens, diseases, heredity, environment and other factors. At present, the clinical methods for the treatment of esophageal cancer mainly include surgical therapy, radiotherapy, chemotherapeutic drug therapy, and non-surgical therapy including endoscopic therapy. However, early diagnosis of esophageal cancer is d...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/158G01N33/57407G01N33/57484G01N33/68
Inventor 杨承刚宋宏涛王晓云
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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