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Gene markers for diagnosis and treatment of tongue squamous cell carcinoma

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

Active Publication Date: 2020-03-31
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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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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  • Gene markers for diagnosis and treatment of tongue squamous cell carcinoma
  • Gene markers for diagnosis and treatment of tongue squamous cell carcinoma
  • Gene markers for diagnosis and treatment of tongue squamous cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1 Screening of differentially expressed genes

[0063] 1. Experimental materials

[0064] Five cases of tongue squamous cell carcinoma tissue specimens were obtained from oral and maxillofacial surgery patients, including 2 cases of well-differentiated squamous cell carcinoma, 2 cases of moderately differentiated squamous cell carcinoma, and 1 case of poorly differentiated squamous cell carcinoma; including 2 male cases and 3 female cases. At the same time, each case of cancerous tissue was selected as the normal tissue at >5cm around the cancerous tumor as its own control. All patients had not undergone chemotherapy, radiotherapy, biological therapy and other treatments for tumors before seeing the doctor. A part of the tissue was immediately stored in liquid nitrogen after collection.

[0065] 2. RNA extraction and cDNA synthesis

[0066] Total RNA was extracted with Trizol RNA reagent (Invitrogen Company), and total RNA was identified by UV spectrophotomet...

Embodiment 2

[0075] Example 2 Verification of Differentially Expressed Genes in Large Samples

[0076] Based on the results of the primary screening of the microarray, we selected the GAGE2D gene for large-scale validation.

[0077] 1. Sample collection

[0078] According to the method of Example 1, 45 tongue squamous cell carcinoma tissues and 45 normal tissues were collected.

[0079] 2. Validation at the mRNA level

[0080] 2.1 Extract tissue RNA

[0081] Step is with embodiment 1.

[0082] 2.2 Reverse transcription

[0083] Reverse transcription using Primescript 1 st strand cDNA synthesis kit kit, the operation steps are as follows:

[0084] (1) Add the following reaction liquid in the microcentrifuge tube, as shown in Table 1:

[0085] Table 1 Reaction liquid

[0086] Reagent dose RNA 2.0μg dNTP 1.0μl Oligo(dT) 2.0μl RNase free dH 2 o

Add to 10.0μl

[0087] (2) Incubate at 70°C for 5 minutes, then rapidly cool to 4°C;

[0088...

Embodiment 3

[0109] Example 3 GAGE2D gene overexpression

[0110] 1. Construction of GAGE2D gene recombinant plasmid

[0111] (1) amplifying the coding sequence of the GAGE2D gene;

[0112] (2) Design amplification primers;

[0113] (3) The amplified GAGE2D gene was connected to the expression vector pcDNA3.0 to construct the pcDNA3.0-GAGE2D recombinant expression vector.

[0114] 2. Culture and transfection of tongue squamous cell carcinoma cells

[0115] 2.1 Cell culture

[0116] Human tongue squamous carcinoma cell line HN4 was cultured in DMEM / F12 medium containing 10% FBS, 100 U / mL penicillin and 100 μg / mL streptomycin. All cells were placed in 5% CO 2 in a 37 °C cell culture incubator.

[0117] 2.2 Cell transfection

[0118] (1) 24h before transfection, the cells were inoculated with 2×10 5 Inoculate each well in a 6-well plate, add DMEM / F12 medium, adhere to the wall overnight, and perform transfection when the cells reach 80-90% confluency.

[0119] (2) Dilute 1 μg of plas...

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Abstract

The invention belongs to the field of biological medicines, and discloses application of a GAGE2D gene used as a diagnosis and treatment marker for tongue squamous carcinoma. An experiment provided by the invention proves that the expression of the GAGE2D gene in normal tissues is significantly different from that in tongue squamous carcinoma tissues. According to the correlation between the GAGE2D gene and the tongue squamous carcinoma, the GAGE2D gene can be used as a novel molecular marker for clinically diagnosing the tongue squamous carcinoma, and can also be used as a novel target of a medicine for treating the tongue squamous carcinoma at the same time.

Description

technical field [0001] The present invention relates to the field of tumor diagnosis, treatment and prognosis prediction, more specifically, the present invention relates to a method for tumor diagnosis and prognosis prediction by means of detecting GAGE2D abnormality; and a tumor therapeutic agent for activating GAGE2D 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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886G01N33/68G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/158G01N33/57407G01N33/68G01N2800/18
Inventor 肖枫常鹏
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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