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Application of c6orf58 gene in preparation of diagnosis and treatment products for tongue squamous cell carcinoma

A technology for tongue squamous cell carcinoma and products, which is applied in the diagnosis and treatment of tongue squamous cell carcinoma with C6orf58 gene, can solve the problems of wide surgical resection, affecting the quality of life of patients, and large lesion area

Active Publication Date: 2019-08-02
青岛山大齐鲁医院(山东大学齐鲁医院(青岛)) +2
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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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  • Application of c6orf58 gene in preparation of diagnosis and treatment products for tongue squamous cell carcinoma
  • Application of c6orf58 gene in preparation of diagnosis and treatment products for tongue squamous cell carcinoma
  • Application of c6orf58 gene in preparation of diagnosis and treatment products for tongue squamous cell carcinoma

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Differential expression of embodiment 1C6orf58 gene

[0061] 1. Experimental materials:

[0062] Tongue squamous cell carcinoma tissue samples were taken from patients with tongue squamous cell carcinoma undergoing radical resection, a total of 50 cases, and normal mucosal tissues were taken from patients with oral trauma, a total of 50 cases, all tissue samples were confirmed by histopathology and signed voluntarily donated for use in this experiment consent. When the samples were taken, the cancer tissue was the central area of ​​the tumor, and the normal mucosa did not contain submucosal epithelial 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.

[0063] 2. Acquisition of RNA from tongue squamous cell carcinoma tissue and normal mucosal tissue

[0064] Total RNA was extracted fr...

Embodiment 2

[0073] Differential expression of embodiment 2C6orf58 protein

[0074] 1. The research object is the same as in Example 1.

[0075]2. Extract tissue total protein

[0076] Follow the instructions of the EpiQuik Tissue / Cell Total Protein Extraction Kit for protein extraction.

[0077] 3. Western blot detection

[0078] The extracted protein was quantitatively subjected to SDS-PAGE electrophoresis, followed by membrane transfer, blocking, primary antibody incubation, secondary antibody incubation, and color development.

[0079] 4. Statistical processing

[0080] The gray value of the protein band was analyzed using Image J software, and the gray value of the C6orf58 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 wh...

Embodiment 3

[0083] Example 3 Inhibition of C6orf58 Gene Expression

[0084] 1. siRNA design and synthesis

[0085] siRNA sequence against C6orf58:

[0086] siRNA-C6orf58:

[0087] The sense strand is 5'-AGUACUUCAACAGAUCAUCAA-3' (SEQ ID NO.7);

[0088] The antisense strand is 5'-GAUGAUCUGUUGAAGUACUUA-3' (SEQ ID NO.8),

[0089] The above siRNA sequences and the negative control siRNA sequence (siRNA-NC) (the negative control siRNA has no homology to the sequence of the C6orf58 gene) were provided by Shanghai Gemma Pharmaceutical Technology Co., Ltd.:

[0090] 2. Culture and transfection of tongue squamous cell carcinoma cells

[0091] 2.1 Cell culture

[0092] Tongue squamous cell carcinoma cell line Tca8113 was inoculated in RPMI-1640 culture medium containing 10% calf serum, 100 μ / ml penicillin, and 100 mg / ml streptomycin, and placed in 37°C, 5% CO 2 Culture in an incubator, and when the cells reach 85% confluency, they are digested and passaged with 0.25% trypsin.

[0093] 2.2 Cel...

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Abstract

The invention discloses applications of C6orf58 gene as an early stage diagnosis molecular marker of tongue squamous carcinoma. It is confirmed by experiments that: compared with normal mucous tissue, C6orf58 gene expression level of tongue squamous carcinoma tissue is increased obviously; and it is confirmed by RNA interface experiment that C6orf58 is capable of influencing proliferation of tongue squamous carcinoma cells. Drugs used for inhibiting C6orf58 gene expression can be developed based on the invention so as to realize clinical prevention and treatment of tongue squamous carcinoma.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of C6orf58 gene in the diagnosis and treatment of squamous cell carcinoma of the tongue. 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 also complex and diverse, involving physical and chemical factors, microbial infection, genetics, and race. Although the etiology and r...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886G01N33/68G01N33/574A61K45/00A61K39/395A61P35/00
CPCA61K39/3955A61K45/00C12Q1/6886C12Q2600/158G01N33/57407G01N33/68
Inventor 杨中军董莉莉杨承刚
Owner 青岛山大齐鲁医院(山东大学齐鲁医院(青岛))
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