Circular RNA molecular marker for laryngeal squamous cell carcinoma, and detection method and application of circular RNA molecular marker
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
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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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