Application of molecular marker to diagnosis and treatment of oral squamous cell carcinoma

A squamous cell carcinoma and oral cavity technology, which is applied in the application field of molecular markers in the diagnosis and treatment of oral squamous cell carcinoma, can solve the problems that the tumor cannot be cured, the operation is difficult to perform, and the curative effect of immunotherapy is not obvious.

Active Publication Date: 2017-05-31
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, radiotherapy and chemotherapy will inevitably cause serious side effects, while surgical treatment has the characteristics of difficult operation, and the defect of simple surgical treatment cannot cure the tumor.
The efficacy of immunotherapy is not obvious

Method used

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  • Application of molecular marker to diagnosis and treatment of oral squamous cell carcinoma
  • Application of molecular marker to diagnosis and treatment of oral squamous cell carcinoma
  • Application of molecular marker to diagnosis and treatment of oral squamous cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 Screening for Gene Markers Related to Oral Squamous Cell Carcinoma

[0061] 1. Sample collection

[0062] 6 cases of surrounding normal mucosa tissue and oral squamous cell carcinoma tissue were collected, all of which were confirmed by pathological diagnosis. All patients did not receive any form of treatment before operation. The surgically resected samples were frozen in liquid nitrogen, and the patients gave informed consent, and all the above-mentioned samples were obtained with the consent of the organizational ethics committee.

[0063] 2. Preparation of RNA samples (operated using QIAGEN tissue RNA extraction kit)

[0064] Take out the tissue samples frozen in liquid nitrogen, put the tissue samples into a pre-cooled mortar for grinding, and extract and isolate RNA according to the instructions in the kit. details as follows:

[0065] 1) Add Trizol and place at room temperature for 5 minutes;

[0066] 2) Add 0.2ml of chloroform, vibrate the centrif...

Embodiment 2

[0079] Example 2 QPCR sequencing to verify differential expression of GDPD2 gene

[0080] 1. Large-sample QPCR verification of differential expression of GDPD2 gene. According to the sample collection method in Example 1, 80 cases of normal mucosal tissues and 80 cases of oral squamous cell carcinoma tissues were selected.

[0081] 2. RNA extraction The steps are as described in Example 1.

[0082] 3. Reverse transcription:

[0083] 1) Reaction system:

[0084]

[0085] 2) Reverse transcription reaction conditions

[0086] According to the reverse transcription reaction conditions in RNA PCR Kit (AMV) Ver.3.0.

[0087] 60min at 42°C, 2min at 99°C, 5min at 5°C.

[0088] 3) Polymerase chain reaction

[0089] 1) Primer design

[0090] QPCR amplification primers were designed according to the coding sequences of GDPD2 gene and GAPDH gene in Genebank, and synthesized by Biomed Biotech. The specific primer sequences are as follows:

[0091] GDPD2 gene:

[0092] The forw...

Embodiment 3

[0105] Example 3 Differential expression of GDPD2 gene in oral squamous cell carcinoma cell lines

[0106] 1. Cell culture

[0107] Oral squamous cell carcinoma cell lines Tca8113 and HN13, and normal mucosal epithelial cell line HIOEC were purchased from the Ninth People's Hospital Affiliated to Shanghai Jiaotong University. The medium of HIOEC is K-SFM; the medium of Tca8113 and HN13 is DMEM; the culture medium containing 10% fetal bovine serum and 1% P / S was incubated at 37°C and 5% CO 2 , Cultivated in an incubator with a relative humidity of 90%. Change the medium once every 2-3 days, and use 0.25% EDTA-containing trypsin for routine digestion and passage.

[0108] 2. Extraction of total cellular RNA

[0109] 1) Stop the culture when the cells reach 80-90% confluency, digest with 0.25% trypsin and collect the cells in 1.5ml EP tubes, add lm1Trizol to each tube and shake slowly to break the cells, place on ice for 10min.

[0110] 2) Remove protein and DNA: add 0.2ml ch...

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Abstract

The invention discloses application of a molecular marker to the diagnosis and the treatment of an oral squamous cell carcinoma. A biomarker is GDPD2; discovered through a qPCR (quantitative Polymerase Chain Reaction) experiment, the expression of a GDPD2 gene in tissue and a cell of the oral squamous cell carcinoma is regulated down; the GDPD2 gene overexpression can inhibit the proliferation of the cell and decrease the attack rate of a carcinoma cell. The invention provides application of an accelerant of the GDPD2 to the diagnosis and treatment of the oral squamous cell carcinoma; meanwhile, the invention also discloses a method for inhibiting the cell from proliferating.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to the application of a molecular marker in the diagnosis and treatment of oral squamous cell carcinoma, and the specific molecular marker is GDPD2. Background technique [0002] Oral squamous cell carcinoma is the most common malignant tumor in and around the oral cavity, and its incidence accounts for more than 80% of the total incidence of oral tumors. It is one of the malignant diseases that deserves great attention and attention. Although the incidence of oral squamous cell carcinoma has declined in developed countries such as Europe and the United States, the incidence is still on the rise worldwide. In recent years, with the continuous improvement of medical level, great progress has been made in the treatment of many malignant tumors, and the 5-year survival rate of patients has been improved through treatment. However, in the treatment of oral squamous cell carcinoma, due to the p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/573G01N33/574A61K48/00A61K38/46A61K45/00A61P35/00
CPCA61K38/465A61K45/00A61K48/005C12Q1/6886C12Q2600/158G01N33/573G01N33/57407G01N33/57484G01N2800/18
Inventor 杨承刚任静程城
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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