Circular RNA molecular marker for laryngeal squamous cell carcinoma, and detection method and application of circular RNA molecular marker
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A technology for molecular markers and laryngeal squamous cell carcinoma, applied in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc., can solve problems such as poor quality of life, financial burden of patients, and unexplained molecular mechanisms
Pending Publication Date: 2021-02-05
FIRST HOSPITAL OF SHANXI MEDICAL UNIV +3
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Problems solved by technology
Although the research on laryngeal squamous cell carcinoma has achieved some results, the molecular mechanism of its pathogenesis is still not elucidated
In particular, the prognosis of patients with advanced laryngeal squamous cell carcinoma is poor, the 5-year survival rate is less than 50%, and the quality of life is also seriously poor, causing serious economic burdens to patients' physiology, psychology and families.
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Embodiment 1
[0082] A circular RNA molecular marker for the detection of laryngeal squamous cell carcinoma, the circular RNA is hsa_circ_0000033, its genome location is: chr1:26584087-26586293, the corresponding linear gene is CEP85 (NM_022778), the circular The nucleotide sequence of the RNA is shown in SEQ ID NO.1.
[0083] A method for detecting the molecular marker hsa_circ_0000033 in laryngeal squamous cell carcinoma, comprising the following steps:
[0084] (1) Clinically collect laryngeal squamous cell carcinoma tissue, weigh the tissue, and extract the total RNA in the tissue;
[0085] (2) reverse transcription of total RNA into cDNA;
[0086] (3) Perform real-time quantitative PCR detection of cDNA by fluorescent dye method, and detect the Ct values of circular RNAhsa_circ_0000033 and internal reference gene 18s in the sample after the reaction;
[0087] (4) According to the Ct value, the level of hsa_circ_0000033 is normalized by the expression level of the internal reference...
Embodiment 2
[0116] The application of siRNA of hsa_circ_0000033 in laryngeal squamous cell carcinoma is used to knock down the expression of hsa_circ_0000033 in laryngeal squamous cell carcinoma cell line, and the siRNA sequence of hsa_circ_0000033 is shown in SEQ ID NO.6-9:
[0117] SEQ ID NO.6, Sense #1: GGAAGAAAGGCUUCAGAAU
[0118] SEQ ID NO.7, Anti-sense #1: AUUCUGAAGCCUUUCUUCC
[0119] SEQ ID NO.8, Sense #2: AGGAAGAAAGGCUUCAGAA
[0120] SEQ ID NO.9, -sense#2: UUCUGAAGCCUUUCUUCCU.
[0121] The specific expression level of hsa_circ_0000033 in human laryngeal squamous cell carcinoma cell lines FD-LSC-1 and AMC-HN-8 was upregulated.
[0122] The siRNA method of transfection hsa_circ_0000033 in laryngeal squamous cell carcinoma cell line comprises the following steps:
[0123] (1) Human laryngeal squamous carcinoma cell lines FD-LSC-1 and AMC-HN-8 were selected and incubated at 37°C and 5% CO 2 Cultivate in an incubator under conditioned conditions, use a medium containing 10% fetal b...
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Abstract
The invention discloses a circular RNA molecular marker for the laryngeal squamous cell carcinoma, and a detection method and application of the circular RNA molecular marker. The nucleotide sequenceof the circular RNA molecular marker has_circ_0000033 for the laryngeal squamous cell carcinoma is disclosed by SEQ ID NO.1. The expression level of the has_circ_0000033 in the carcinoma tissues of alaryngeal squamous cell carcinoma patient is up-regulated, and the has_circ_0000033 is applied to preparation of an auxiliary diagnosis kit for the laryngeal squamous cell carcinoma. The detection method, which is provided by the invention, of a laryngeal squamous cell carcinoma molecular marker quantity carries out real-time quantitative PCR detection by a fluorescent dye method, and a back-to-back primer designed by aiming at the detection method is disclosed by SEQ ID NO.2-3. The invention provides a specific siRNA sequence of the targeted has_circ_0000033, and the siRNA is used for knocking out has_circ_0000033 expression in a laryngeal squamous cell carcinoma cell line and performing a biological function of inhibiting the multiplication capacity of the laryngeal squamous cell carcinoma cell line. The laryngeal squamous cell carcinoma molecular marker has_circ_0000033 disclosed by the invention has an application prospect of simply and quickly making a laryngeal squamous cell carcinoma auxiliary diagnosis. The siRNA sequence designed by the invention can effectively lower the expression quantity of the has_circ_0000033, and has the characteristics of being specific and quick in acting.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to a circular RNA molecular marker for laryngeal squamous cell carcinoma and its detection method and application. Background technique [0002] Laryngeal squamous cell carcinoma is an aggressive head and neck malignancy derived from the epithelial tissue of the larynx, accounting for approximately 2.4% of all tumors worldwide each year. In recent years, due to factors such as smoking and excessive drinking, the incidence of laryngeal squamous cell carcinoma has increased slightly year by year. Laryngeal squamous cell carcinoma is a malignant tumor of the head and neck with a high incidence in northern my country, with high morbidity, disability and mortality. Although some achievements have been made in the study of laryngeal squamous cell carcinoma, the molecular mechanism of its pathogenesis has not yet been elucidated. In particular, the prognosis of patients with advanced larynge...
Claims
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