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Application of non-coding RNA relevant to occurrence and development of laryngeal squamous cell carcinoma

A technology of laryngeal squamous cell carcinoma and reagents, applied in the application field of non-coding RNA

Inactive Publication Date: 2019-08-23
THE SECOND HOSPITAL OF HEBEI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the diagnosis of laryngeal cancer still relies on traditional symptoms, signs, laryngoscopy, CT manifestations and pathological diagnosis. There is no effective serum tumor marker that can be widely used in clinical practice for the screening and early diagnosis of laryngeal cancer.

Method used

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  • Application of non-coding RNA relevant to occurrence and development of laryngeal squamous cell carcinoma
  • Application of non-coding RNA relevant to occurrence and development of laryngeal squamous cell carcinoma
  • Application of non-coding RNA relevant to occurrence and development of laryngeal squamous cell carcinoma

Examples

Experimental program
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Effect test

Embodiment approach

[0033] The invention provides a product for diagnosing laryngeal squamous cell carcinoma, the product includes a reagent for detecting the expression level of LOC100507599, and the product includes a chip or a kit. As an embodiment, the chip includes a solid phase carrier and an oligonucleotide probe that specifically recognizes LOC100507599 immobilized on the solid phase carrier, and the kit includes primers that specifically amplify LOC100507599 and oligonucleotides that specifically recognize LOC100507599 probe or chip.

[0034] The oligonucleotide probe specifically recognizing LOC100507599 may be DNA, RNA, DNA-RNA chimera, PNA or other derivatives. The length of the probe is not limited, as long as it completes specific hybridization and specifically binds to the target nucleotide sequence, any length is acceptable. The probes can be as short as 25, 20, 15, 13 or 10 bases in length. Likewise, the probes can be as long as 60, 80, 100, 150, 300 base pairs or longer, or ev...

Embodiment 1

[0043] Example 1 Screening for gene markers associated with laryngeal squamous cell carcinoma

[0044] 1. Sample collection

[0045] The cancer tissues and corresponding paracancerous tissue samples (more than 1.5 cm from the tumor edge) of 4 cases of laryngeal squamous cell carcinoma were collected for high-throughput sequencing. All patients had not received chemotherapy, radiotherapy and endocrine therapy before operation, and the patients had no other Tumor, cardiovascular and cerebrovascular disease history, infectious disease history and family hereditary disease history.

[0046] 2. RNA sample preparation and quality analysis

[0047] Use the Takara RNA Extraction Kit (Code NO.9767) to extract RNA from the tissue, the steps are as follows:

[0048] 1) Transfer the tissue sample to a liquid nitrogen pre-cooled mortar, grind the tissue with a pestle, and add liquid nitrogen continuously until it is ground into powder. Add the powdered sample into a 1.5ml sterilized centr...

Embodiment 2

[0070] Example 2 QPCR sequencing verification of differential expression of LOC100507599 gene

[0071] 1. The differential expression of the LOC100507599 gene was verified by large sample QPCR of the cancer tissue samples and para-cancerous tissue samples collected from 32 patients with laryngeal squamous cell carcinoma according to the collection method in Example 1.

[0072] 2. RNA extraction

[0073] Takara RNA Extraction Kit (Code NO.9767) was used to extract RNA from tissue, see Example 1 for specific steps.

[0074] 3. QPCR

[0075] Primers were designed according to the gene sequences of LOC100507599 and GADPH, and the primer sequences are shown in Table 1.

[0076] Table 1 Amplification Primer Sequence

[0077]

[0078]

[0079] Using TaKaRa One Step TB Green TM Prime Script TM The RT-PCR kit (Code No. RR066A) was used for PCR reaction, and the reaction system and reaction conditions are shown in Table 2. In Thermal Cycler Perform PCR amplification on t...

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Abstract

The invention discloses application of a non-coding RNA relevant to occurrence and development of laryngeal squamous cell carcinoma. The non-coding RNA is LOC100507599. According to the application, through combination of a high throughput sequencing technology with bioinformation analysis, it is found for the first time that expression of the LOC100507599 in a patient suffering from the laryngealsquamous cell carcinoma is up-regulated, and the conclusion is verified through further QPCR, which indicates that the LOC100507599 can serve as a molecular target to be applied to diagnosis and treatment of the laryngeal squamous cell carcinoma.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to the application of a non-coding RNA related to the occurrence and development of laryngeal squamous cell carcinoma, and the specific RNA involved is LOC100507599. Background technique [0002] Laryngeal cancer is the most common malignant tumor of the larynx. In my country, the Northeast region has the highest incidence rate. In recent years, the incidence rate has increased significantly. However, the pathogenesis of laryngeal cancer has not been fully elucidated so far. At present, the treatment of laryngeal cancer is mainly surgical resection. After treatment (including total laryngectomy and partial laryngectomy), the 5-year survival rate of stage I is about 94%, stage II is about 89%, and stage III is about 82%. , Stage IV is about 66%. Early laryngeal cancer can be cured by surgery or radiotherapy; patients with advanced laryngeal cancer usually have severely affected pronunciatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/113
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 张海中尹晓妍王静妙
Owner THE SECOND HOSPITAL OF HEBEI MEDICAL UNIV
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