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Application of LncRNA IFITM4P as molecular marker in oral leukoplakia and/or oral cancer

A technology of molecular markers and oral leukoplakia, applied in the field of biomedical detection, can solve the problems of oral leukoplakia recurrence, multiple cancers, multiple occurrences, etc., and achieve the effect of improving low sensitivity

Pending Publication Date: 2022-06-17
SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the local and systemic treatment of drugs and surgery, laser, cryotherapy, photodynamic therapy and other therapies cannot effectively reduce the canceration rate of oral leukoplakia, and oral leukoplakia still has clinical difficulties such as easy recurrence, multiple occurrences, and multiple cancerations
[0004] Long non-coding RNAs (long non-coding RNAs, lncRANs) are RNA transcripts >200 nt in length, lacking significant protein-coding potential

Method used

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  • Application of LncRNA IFITM4P as molecular marker in oral leukoplakia and/or oral cancer
  • Application of LncRNA IFITM4P as molecular marker in oral leukoplakia and/or oral cancer
  • Application of LncRNA IFITM4P as molecular marker in oral leukoplakia and/or oral cancer

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] RNA extraction and RT-PCR

[0092] (1) RNA extraction steps

[0093] a) When extracting tissue RNA, use 1 ml of Trizol reagent per 50-100 mg of tissue to dissolve the tissue, add 2-3 iron beads to the 1.5 ml EP tube added to the tissue, and use a cryogenic freezer automatic grinder to break the tissue; extract cells For RNA, centrifuge the cells at 800 g for 5 min, add 1 ml of Trizol to every 5 to 106 cells, and mix with a 1 ml pipette to lyse the cells.

[0094] b) The above-mentioned Trizol lysis solution of the tissue or cells was placed at room temperature for 5 minutes to fully lyse the tissue and cells.

[0095] c) In the above EP tube, add chloroform (trichloromethane) in an amount of 0.2 ml per 1 ml of Trizol, cover the EP tube, shake it on a vortex shaker for 15 seconds, and place it at room temperature (about 22°C). After 3 minutes, centrifuge at 12000g for 15 minutes at 4°C to separate the organic phase and the RNA-containing aqueous phase.

[0096] d) Tra...

Embodiment 2

[0127] To identify differentially expressed lncRNAs in oral normal mucosa (NM), OL and OSCC, in this example NM (n=3), OL (n=4) and OSCC (n=5) samples from Chinese patients were subjected to microarray analysis. array analysis, such as figure 1 A and Table 1.

[0128] The analysis revealed 3109 interactions between lncRNAs and mRNA transcripts in the NM, OL and OSCC groups (p2); focusing on 10 differentially expressed lncRNAs (5 genes were up-regulated, 5 genes were down-regulated), the difference fold>2, ***pfigure 1 C). The expression of IFITM4P was highest in OSCC / OL and OL / NM groups. To validate these observations, qRT-PCR was used to detect the expression of IFITM4P in samples of oral normal mucosa (NM) (n=23), OL (n=64) and OSCC (n=43), respectively, in contrast to OL and NM ratio, IFITM4P has the highest expression in OCSS and the lowest expression in NM ( figure 1 D), These results were validated by RNA fluorescence in situ hybridization (FISH) staining of samples ...

Embodiment 3

[0133] To determine the role of IFITM4P in oral carcinogenesis, Leuk-1 (OL) and HN4 (OSCC) cells were selected for this experiment, and were manipulated by stably transducing Leuk-1 cells with a lentiviral vector carrying a cDNA encoding the full length of IFITM4P Expression of IFITM4P ( figure 2 A), and analyzed cell growth and colony formation in these cells. The results showed that exogenous IFITM4P increased cell growth and colony formation in Leuk-1 cells ( figure 2 B, 2D and 2E).

[0134] In order to verify the opposite results, the present invention depleted IFITM4P in Leuk-1 cells by using IFITM4P-specific short hairpin RNA (shRNA) ( figure 2 A), and observed a significant reduction in cell growth and colony formation ( figure 2 B, 2D and 2E). Similar results were observed in the high expression of IFITM4P ( figure 2 F, 2G, 2I, and 2J) and deletions ( figure 2 F, 2H, 2I and 2J) were also seen in HN4 cells. In order to study the effect of IFITM4P in OSCC in...

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Abstract

The invention discloses an application of LncRNA IFITM4P as a molecular marker in oral leukoplakia and / or oral cancer. The nucleotide sequence of the LncRNA IFITM4P is shown as Seq ID NO: 1. According to the LncRNA IFITM4P related to the oral cancer, provided by the invention, the LncRNA IFITM4P is highly expressed in a tissue sample of a patient with the oral cancer, and the LncRNA IFITM4P can be used as a molecular marker of the oral leukoplakia and / or the oral cancer and can be applied to a product for diagnosis and / or curative effect monitoring of the oral leukoplakia and / or the oral cancer.

Description

technical field [0001] The invention relates to the technical field of biomedical detection, in particular to the application of LncRNA IFITM4P as a molecular marker in oral leukoplakia and / or oral cancer. Background technique [0002] Oral cancer (OSCC) is a common malignant tumor with poor prognosis, with more than 300,000 new cases worldwide every year. Oral precancerous lesions refer to certain clinical and histological changes in the oral and maxillofacial region with a tendency to cancer, including leukoplakia, erythema, lichen planus, discoid lupus erythematosus, submucosal fibrosis, papilloma, Chronic ulcers, mucosal black spots and pigmented nevus, etc. Among them, oral leukoplakia (OL) is the most typical type of oral potential malignant disorder (OPMD). Currently, effective markers and their mechanisms for predicting oral carcinogenesis are still unknown. [0003] Among them, oral leukoplakia (OL) is one of the most important potential malignant diseases of ora...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11C12Q1/6851
CPCC12Q1/6886C12Q1/6851C12Q2600/178C12Q2600/158C12Q2561/113C12Q2531/113C12Q2521/107
Inventor 施琳俊
Owner SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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