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Laryngeal squamous cell carcinoma molecular marker hsa_circ_0004547 and detection method and application thereof

A molecular marker, laryngeal squamous cell carcinoma technology, applied in biochemical equipment and methods, microbial determination/inspection, medical preparations containing active ingredients, etc., can solve problems such as off-target effects and unreasonable design, and achieve detection efficiency high effect

Inactive Publication Date: 2020-06-26
FIRST HOSPITAL OF SHANXI MEDICAL UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Small interfering RNA (siRNA), known as short interfering RNA or silencing RNA, is a double-stranded RNA of 20 to 25 nucleotides long and has many different uses in biology. It interferes with expression and complementary nucleotides The post-transcriptional degraded RNA of a specific gene sequence, this method is simple and efficient, and has gradually become a powerful gene silencing method in molecular cell biology, but if the design is not reasonable, it is very easy to produce off-target effects

Method used

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  • Laryngeal squamous cell carcinoma molecular marker hsa_circ_0004547 and detection method and application thereof
  • Laryngeal squamous cell carcinoma molecular marker hsa_circ_0004547 and detection method and application thereof
  • Laryngeal squamous cell carcinoma molecular marker hsa_circ_0004547 and detection method and application thereof

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Embodiment 1

[0045] A detection method and application of the molecular marker hsa_circ_0004547 and its expression in laryngeal squamous cell carcinoma.

[0046] A molecular marker for laryngeal squamous cell carcinoma. The molecular marker for laryngeal squamous cell carcinoma is hsa_circ_0004547, and its genome location is: chr22:29517344-29521404. The nucleotide sequence of hsa_circ_0004547 is shown in SEQ ID NO:1.

[0047] The expression level of hsa_circ_0004547 in laryngeal cancer patients is up-regulated. The malignancy of laryngeal squamous cell carcinoma is correlated with the hsa_circ_0004547 molecular expression, the higher the malignancy, the higher the hsa_circ_0004547 molecular expression.

[0048] The laryngeal squamous cell carcinoma molecular marker hsa_circ_0004547 is used in the preparation of an auxiliary diagnostic kit for laryngeal squamous cell carcinoma.

[0049] A method for detecting the expression level of the laryngeal cancer molecular marker hsa_circ_0004547, co...

Embodiment 2

[0079] The siRNA of hsa_circ_0004547 was synthesized by using the laryngeal cancer molecular marker hsa_circ_0004547 of the present invention.

[0080] First, siRNA for hsa_circ_0004547 was synthesized.

[0081] According to the siRNA design principle, the synthetic siRNA sequence contains the circularization site of the circular RNA, and the sequence with a fragment length of 20 bp is used as the candidate target sequence. By comparing the human genome sequence, it is ensured that there is no homologous sequence. The siRNA designed in the present invention The sequence is as follows:

[0082] Sense strand: 5'-UCUUUGAUAUGCCUGGAAA-3'

[0083] Antisense strand: 5'-UUUCCAGGCAUAUCAAAGA-3'

[0084] Send the sequence to Suzhou Gemma Company for synthesis, add two deoxyribonucleic acid dTdT, increase the stability of siRNA, and label siRNA with green fluorescence.

[0085] Second, siRNA transfection was performed on laryngeal squamous cell carcinoma cells.

[0086] Laryngeal squa...

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Abstract

The present invention discloses a laryngeal squamous cell carcinoma molecular marker hsa_circ_0004547 and a detection method and an application thereof, and belongs to the technical field of biology.A location of the provided laryngeal squamous cell carcinoma molecular marker hsa_circ_0004547 on the genome is: chr22:29517344-29521404 and has a nucleotide sequence shown in SEQ ID NO:1. The expression level of the hsa_circ_0004547 is up-regulated in tissues of patients with laryngeal squamous cell carcinoma. Back-to-back primers of the hsa_circ_0004547 in real-time fluorescence quantification in a method for detecting the expression of the laryngeal squamous cell carcinoma molecular marker are F1: 5'-GGCAGGAATTGCAGCGAGAA-3'; and R1: 5'-AAAGCAGCAGGATGGTGGC-3'. The present invention providesa siRNA sequence of the hsa_circ_0004547. The siRNA of the hsa_circ_0004547 is used to knock down the expression of the hsa_circ_00004547 in laryngeal squamous cell carcinoma cells, has an inhibitoryeffect on migration ability of the laryngeal squamous cell carcinoma cells, and can be used to prepare chemotherapy drugs for the laryngeal squamous cell carcinoma. The screened out hsa_circ_0004547 can be used as a new molecular marker for an early diagnosis of the laryngeal squamous cell carcinoma and detection of the molecular marker can be used for simple and quick initial diagnosis of the laryngeal squamous cell carcinoma. The designed siRNA can effectively reduce the expression level of the hsa_circ_0004547 and has the characteristics of high efficiency and rapid action.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a molecular marker hsa_circ_0004547 for laryngeal squamous cell carcinoma, a detection method and an application. Background technique [0002] Laryngeal squamous cell carcinoma, as the second most common tumor in head and neck tumors, is an aggressive tumor, which accounts for about 2.4% of emerging malignant tumors in the world every year, and its mortality rate accounts for 1% of cancer deaths worldwide. There are 10,000 people diagnosed with laryngeal squamous cell carcinoma, and the morbidity and mortality have been increasing in recent years. The treatment of laryngeal squamous cell carcinoma mainly relies on functional laryngeal surgery, radiotherapy and chemotherapy. More and more studies focus on improving the clinical effect of laryngeal squamous cell carcinoma by identifying biomarkers, but the drug resistance and recurrence of the tumor The prognosis of patients with lar...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/113A61K48/00A61K31/713A61P35/00
CPCA61K31/713A61P35/00C12Q1/6886C12Q2600/118C12Q2600/158C12Q2600/178
Inventor 高伟吴勇延薛绪亭张宇良郑希望牛敏代力代丰升侯睿韩琦
Owner FIRST HOSPITAL OF SHANXI MEDICAL UNIV
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