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Application of miR-1207 and target gene thereof in detection of laryngeal squamous cell carcinoma

A technology of mir-1207 and laryngeal squamous cell carcinoma, applied in the application field of miR-1207 and its target gene in the detection of laryngeal squamous cell carcinoma

Active Publication Date: 2021-03-26
SHENZHEN HOSPITAL OF SOUTHERN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is currently no standard treatment strategy that can be used to improve the survival rate of patients, especially those with advanced laryngeal cancer

Method used

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  • Application of miR-1207 and target gene thereof in detection of laryngeal squamous cell carcinoma
  • Application of miR-1207 and target gene thereof in detection of laryngeal squamous cell carcinoma
  • Application of miR-1207 and target gene thereof in detection of laryngeal squamous cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Screening of biomarkers associated with laryngeal squamous cell carcinoma

[0052] 1. Sample collection

[0053] Six cases of laryngeal squamous cell carcinoma tissues and corresponding paracancerous tissue samples were collected for high-throughput sequencing.

[0054] 2. RNA sample preparation and quality analysis

[0055] Use the TRIZOL method to extract tissue RNA, the steps are as follows:

[0056] 1) Pre-cool the mortar with liquid nitrogen, put the tissue sample into the mortar with liquid nitrogen, and fully grind the tissue sample into powder under liquid nitrogen.

[0057] 2) Transfer the sample powder into a 2.0mL EP tube filled with TRIzol lysate, shake vigorously, mix well, and let it stand at room temperature for 5-10min.

[0058] 3) Centrifuge at 10000 rpm at 4°C for 5 minutes.

[0059] 4) Pipette the supernatant into a new 2.0mL EP tube, add 200μL of chloroform / isoamyl alcohol to each mL of the lysate, and mix vigorously by inversion.

[0...

Embodiment 2

[0097] Example 2 QPCR detection of miRNA target gene expression in laryngeal squamous cell carcinoma samples

[0098] 1. Carcinoma tissue samples and paracancerous tissue samples collected from 29 patients with laryngeal squamous cell carcinoma according to the collection method in Example 1 were subjected to large sample QPCR verification of differentially expressed genes.

[0099] 2. Extraction of RNA

[0100] Use the TRIZOL method to extract tissue RNA, and the specific steps are the same as Example 1.

[0101] 3. cDNA synthesis by reverse transcription

[0102] 3.1 Synthesis of lncRNA cDNA by reverse transcription

[0103] mRNA reverse transcription was performed using the FastQuant cDNA First Strand Synthesis Kit (Product No.: KR106).

[0104] 1) To remove the genomic DNA reaction, add 2.0 μl of 5×gDNA Buffer, 1 μg of TotalRNA, and RNase Free ddH to the test tube 2 O to bring total volume to 10 μl.

[0105] 2) Heat in a water bath at 42°C for 3 minutes.

[0106] 3) ...

Embodiment 3

[0126] Example 3 Silencing of Genes and Its Effects on Laryngeal Squamous Carcinoma Cells

[0127] 1. Cell culture

[0128] The human laryngeal squamous cell line Hep2 was cultured in RPMI1640 medium (containing 10% fetal bovine serum) at 37°C, 5% CO 2 Routine culture and subculture under saturated humidity conditions.

[0129] 2. Transfection

[0130] Hep-2 cells in the logarithmic growth phase were inoculated into 6-well plates, and transfected in groups when the cell confluence reached 80% to 90%. The transfection reagent used Lipofectamine 2000. The experiment was divided into three groups: blank control group (Hep-2); negative control group (transfected with universal siRNA-NC); experimental group: transfected with siRNA.

[0131] The siRNA interference fragments targeting LOC339685 and LOC400706 genes were designed and synthesized by Shanghai Jima Pharmaceutical Technology Co., Ltd. (as shown in Table 3).

[0132] Table 3 siRNA sequence

[0133]

[0134] 3. QPCR de...

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Abstract

The invention discloses application of miR-1207 and a target gene thereof in detection of laryngeal squamous cell carcinoma. The target gene is LOC339685 or LOC400706. The invention discloses application of a reagent for detecting LOC339685 or LOC400706 in preparation of a product for diagnosing laryngeal squamous cell carcinoma, and also discloses application of LOC339685 or LOC400706 in preparation of a medicine for treating laryngeal squamous cell carcinoma.

Description

technical field [0001] The invention relates to the field of molecular diagnosis, in particular to the application of miR-1207 and its target gene in detecting laryngeal squamous cell carcinoma. Background technique [0002] Laryngeal cancer is one of the common malignant tumors in the head and neck, and 96-98% of the pathological types are laryngeal squamous cell carcinoma (LSCC). According to the latest relevant data compiled by the International Agency for Research on Cancer, there were 177,400 new laryngeal cancer cases and 94,800 deaths worldwide in 2018 (Bray F, FerlayJ, Soerjomataram I, et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J].CA CancerJ C1in,2018,68(6):394-424.); and the latest domestic statistics show that in 2015, the number of new cases of laryngeal cancer in my country was 26,400 cases, and the number of deaths was 14,500 (Chen W, Zheng R, Baade PD, et al. Cancer statistics in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11G16B30/00G16B20/00A61K31/713A61K45/00A61P35/00
CPCC12Q1/6886G16B30/00G16B20/00A61K31/713A61K45/00A61P35/00C12Q2600/158C12Q2600/178
Inventor 崔晓峰陈向军张思林
Owner SHENZHEN HOSPITAL OF SOUTHERN MEDICAL UNIV
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