CDSN as diagnosis and treatment target of tongue squamous carcinoma

A tongue squamous cell carcinoma and product technology, applied in the field of tumor diagnosis and prognosis prediction, can solve the problems of large lesions, affecting the quality of life of patients, and wide surgical resection.

Active Publication Date: 2017-06-30
GENERAL HOSPITAL OF PLA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the size of the lesion is large and the scope of surgical resection is wide, which will seriously affect the quality of life of patients after surgery.

Method used

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  • CDSN as diagnosis and treatment target of tongue squamous carcinoma
  • CDSN as diagnosis and treatment target of tongue squamous carcinoma
  • CDSN as diagnosis and treatment target of tongue squamous carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1 Screening of differentially expressed genes

[0063] 1. Materials:

[0064] Tongue squamous cell carcinoma and paracancerous tissue specimens were taken from patients with tongue squamous cell carcinoma undergoing radical resection. There were 5 patients. All tissue samples were confirmed by histopathology, and a consent form for voluntary donation for use in this experiment was signed. The cancerous tissue was the central area of ​​the tumor when the material was collected; the corresponding paracancerous tissue was the tissue 1-2cm away from the edge of the cancerous tissue. All specimens were placed in 1.5ml EP tubes and stored in liquid nitrogen tanks. All specimens were collected from patients who had not undergone any form of anti-tumor treatment and had no history of tumors in other locations.

[0065] 2. Obtaining tissue RNA

[0066] Total RNA was extracted from tissue samples, and the concentration and purity of the extracted RNA were detected by N...

Embodiment 2

[0092] Example 2 Large sample verification screened out differentially expressed genes

[0093] Based on the results of the previous high-throughput transcriptome deep sequencing, and according to the size of the P value, we selected the CDSN gene for verification.

[0094] 1. Sample collection

[0095] According to the method of Example 1, 45 tongue squamous cell carcinoma tissues and 45 corresponding paracancerous tissues were collected.

[0096] 2. Validation at the mRNA level

[0097] 2.1 Extract tissue RNA

[0098] Step is with embodiment 1.

[0099] 2.2 Reverse transcription

[0100] Reverse transcription using Primescript 1 st strand cDNAsynthesis kit kit, the operation steps are as follows:

[0101] (1) Add the following reaction liquid in the microcentrifuge tube, as shown in Table 1:

[0102] Table 1 Reaction liquid

[0103]

[0104]

[0105] (2) Incubate at 70°C for 5 minutes, then rapidly cool to 4°C;

[0106] Add the following reagents into a micro...

Embodiment 3

[0127] Example 3 CDSN gene overexpression

[0128] 1. CDSN gene recombinant plasmid construction

[0129] (1) amplifying the coding sequence of the CDSN gene;

[0130] (2) Design amplification primers;

[0131] (3) Link the amplified CDSN gene into the expression vector pcDNA3.0.

[0132] 2. Culture and transfection of tongue squamous cell carcinoma cells

[0133] 2.1 Cell culture

[0134] Tongue squamous carcinoma cell line Tca8113 was inoculated in RPMI-1640 medium containing 10% calf serum, 100 μ / ml penicillin, and 100 mg / ml streptomycin.

[0135] 2.2 Cell transfection

[0136] (1) The day before transfection, 1*10 5 Tumor cells were suspended in 500 μl of antibiotic-free medium and seeded into 24-well culture plates.

[0137] (2) Transfect when the cell density reaches 85%. Dilute 1 μg of recombinant plasmid DNA in serum-free medium and mix gently; take 4 μl Lipofectamine2000 and dilute it in serum-free medium, mix well, and mix the two parts Mix and stand for 20 m...

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Abstract

The invention discloses a CDSN gene as a diagnosis and treatment target of tongue squamous carcinoma. By detecting content of CDSN gene and its expression product in tissues of a subject, whether the subject is affected with tongue squamous carcinoma can be judged or whether the subject has the risk of suffering from tongue squamous carcinoma can be diagnosed, or a patient's reactivity to drug therapy can be judged, or a patient's relapse and prognosis can be judged. In addition, through the research on proliferation index of tongue squamous carcinoma cells cultured in vitro, it is proved that the CDSN gene and its expression production are used as the drug target for treatment of tongue squamous carcinoma.

Description

technical field [0001] The present invention relates to the fields of tumor diagnosis, treatment and prognosis prediction, more specifically, the present invention relates to a method for tumor diagnosis and prognosis prediction by detecting CDSN abnormality; and a tumor therapeutic agent for activating CDSN gene or protein. Background technique [0002] Oral squamous cell carcinoma is the most common malignant tumor of the head and neck, accounting for about 3% of systemic tumors and 90% of oral tumors, and tongue cancer ranks first in the incidence of oral squamous cell carcinoma. Although the incidence of oral cancer in developed countries has shown a slow downward trend in recent years, the overall incidence is increasing. Most of the patients are men over 40 years old, and the predilection sites are mostly on the tongue, cheeks, and gums. The occurrence of tongue squamous cell carcinoma is a multi-factor, multi-step, and multi-stage complex process, and the etiology is...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574
CPCC12Q1/6886C12Q2600/118C12Q2600/158G01N33/57407G01N33/68G01N2333/47
Inventor 汪林姜华白阳杨承刚
Owner GENERAL HOSPITAL OF PLA
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