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Application of miR-518a-3p in diagnosis of oral carcinoma

A mir-518a-3p, oral cancer technology, applied in medical preparations containing active ingredients, determination/inspection of microorganisms, DNA/RNA fragments, etc. The family survival status and quality of life decline of patients with squamous cell carcinoma

Inactive Publication Date: 2018-01-19
XIANGYA STOMATOLOGICAL HOSPITAL CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Surgical resection is still the main treatment, although the progress of microvascular free flap reconstruction has broken many resection restrictions, and can restore the function and appearance to the maximum extent, but due to the particularity of the site of oral squamous cell carcinoma, oral squamous cell carcinoma Pronunciation, eating, and social interaction of cancer patients have been seriously affected after surgery
And because the postoperative labor ability of patients with oral squamous cell carcinoma is seriously affected, the family survival status and quality of life of patients with oral squamous cell carcinoma have declined sharply

Method used

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  • Application of miR-518a-3p in diagnosis of oral carcinoma
  • Application of miR-518a-3p in diagnosis of oral carcinoma
  • Application of miR-518a-3p in diagnosis of oral carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Sample collection, sequencing and data analysis

[0050] The samples of the 6 cases of oral squamous cell carcinoma tissues and their adjacent normal tissues were all surgically resected specimens in the hospital, and all the specimens were placed in a liquid nitrogen tank within 10 minutes of isolation, and then transferred to a -80°C refrigerator for storage. Sent to the company for analysis of miRNA chip data, and screened out miRNAs with significant differential expression. By performing log2 processing on the Ford Change value of miRNA, and performing -log10 processing on the corresponding p value, a volcano map is drawn (see figure 1 ). In the analysis results, there were 55 miRNAs with p<0.05, among which 38 were up-regulated and 17 were down-regulated. In addition to several miRNAs that have been reported in the literature and are closely related to oral squamous cell carcinoma (such as miR-125b, miR-145, miR-21, miR-181b, miR-362-3p, miR-99a, let-7e...

Embodiment 2

[0051] Example 2 Cell Culture

[0052] The human oral squamous cell carcinoma cell lines UM1 and UM2 used in the experiment were donated by Guanghua School of Stomatology, Sun Yat-sen University. The human oral squamous cell carcinoma cell lines SCC9 and SCC15 were purchased from ATCC (American Type Culture Collection). The cell lines CAL27 and Tca8113 were donated by the Cancer Research Institute of the Affiliated Cancer Hospital of Guangzhou Medical University. The above cells were used with 10% fetal bovine serum, 1×10 5 U / L penicillin and 1×10 5 Cultured in U / L streptomycin DMEM / F12 (1:1) complete medium at 37°C, 5% CO 2 and cultured under saturated humidity conditions, subcultured once every 2-3 days, and the cells in the exponential growth phase were used for experiments. HaCat is human normal epidermal cells purchased from the Institute of Basic Medical Sciences, Chinese Academy of Sciences, cells containing 10% fetal bovine serum, 1 × 10 5 U / L penicillin and 1×10 5...

Embodiment 3

[0061] The extraction of embodiment 3miRNA

[0062] 3.1 Extraction of tissue miRNA

[0063] The miRNAs extraction process was carried out according to the instructions of The RNeasy Plus Mini Kit (QIAGEN, Germany). Specific steps are as follows:

[0064] 1) Wash the mortar and grinding rods under running water, air-dry them, wrap them in tin foil, and place them in a high-temperature oven at 180°C for 12 hours to inactivate the enzymes. After taking it out, cool it at room temperature, pour a small amount of liquid nitrogen into the mortar to keep the temperature of the mortar below 4°C. Put the frozen tissue fragments into a mortar, add liquid nitrogen to make the tissue brittle, grind it with a grinding rod, grind it repeatedly to crush the tissue, and transfer the powdered tissue to a dry Ep tube.

[0065] 2) Add 70 μl QIA20l Lysis Reagent to the Ep tube, pipette, mix for 1 min, and place at room temperature for 5 min.

[0066]3) Add 140 μl of chloroform, seal, shake fo...

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Abstract

The invention relates to an application of miR-518a-3p in diagnosis of oral carcinoma, in particular to the application of miR-518a-3p and a precursor of miR-518a-3p in preparation of oral squamous cell carcinoma diagnosis preparations. MiRNA microarray data analysis is performed on oral squamous cell carcinoma and adjacent normal tissue with an SAM (selectively amplified microsatellite) method. MiRNA with significant differential expression is screened out, and miR-518a-3p as a novel molecular diagnostic marker associated with the oral squamous cell carcinoma is found. Further, experimental results of fluorescence quantitative PCR verify high expression of miR-518a-3p in oral squamous cell carcinoma tissue and the cell line of the oral squamous cell carcinoma. The novel molecular marker is provided for accurate clinical diagnosis of the oral carcinoma, and a foundation is laid for subsequent development of oral squamous cell carcinoma molecular diagnosis products.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to early molecular diagnostic markers for oral cancer and their application in clinical diagnosis and treatment, and more specifically to the application of miR-518a-3p and its precursors in the preparation of oral squamous cell carcinoma diagnostic preparations. Background technique [0002] In the past 20 years, with the advancement of science and technology, the continuous improvement and updating of tumor detection, treatment methods and equipment, the five-year survival rate of oral squamous carcinoma has improved by 5%, but compared with other tumors in the past 20 years The five-year survival rate has increased, but the five-year survival rate for oral squamous cell carcinoma is still around 50%, which is at a relatively low level. Surgical resection is still the main treatment, although the progress of microvascular free flap reconstruction has broken many resection restriction...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11A61K45/00A61P35/00
Inventor 潘灏唐瞻贵王月红刘斌杰白新娜刘欧胜全宏志周玥颖方小丹王柏胜邓智元顾立群
Owner XIANGYA STOMATOLOGICAL HOSPITAL CENT SOUTH UNIV
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