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Application of methylation detection of cancer suppressor gene promoter in assessing oral leukoplakia canceration

A technology of oral leukoplakia and tumor suppressor genes, which is applied in the field of epigenetics to achieve stable detection results, good repeatability, and improved specificity and sensitivity

Inactive Publication Date: 2011-09-21
SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At the same time, there are some patients with leukoplakia cancer in clinic, and there is no abnormal epithelial hyperplasia in the lesion tissue slices before leukoplakia. This phenomenon poses a challenge to the current gold standard for predicting leukoplakia carcinogenesis.

Method used

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  • Application of methylation detection of cancer suppressor gene promoter in assessing oral leukoplakia canceration
  • Application of methylation detection of cancer suppressor gene promoter in assessing oral leukoplakia canceration
  • Application of methylation detection of cancer suppressor gene promoter in assessing oral leukoplakia canceration

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Embodiment 1

[0025] I. Tissue sample DNA bisulfite treatment:

[0026] (1) After clinically obtaining the tissue, 25 frozen sections of 20um tissue were taken and digested overnight at 48°C with 1% proteinase K (Merck). The DNA was extracted by phenol-chloroform method the next day and dissolved in 100ulTE solution.

[0027] (2) 2ug extracted DNA samples were mixed with 0.2mol / L NaOH solution, denatured at 50°C for 20min. The denatured DNA was incubated in 500ul freshly prepared bisulfite solution (2.5M bisulfite, 125mM hydroquinone and 3M NaOH mixture) at 70°C for 3h. The treated DNA was purified with a Wizard Clean-up kit (Promega), and after 10 min of 0.3 mol / L NaOH solution, 1 ul glycogen and 350 ul ethanol were added to precipitate the DNA overnight.

[0028] (3) Centrifuge at 13000rmp for 15min to obtain bisulfite-treated DNA, which is dissolved in 60ul double-distilled water for detection.

[0029] II. Quantitative methylation PCR to measure gene promoter methylation expression: ...

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Abstract

The invention discloses an application of a methylation detection of a cancer suppressor gene promoter in assessing oral leukoplakia canceration. By quantitatively detecting the methylation expression of a cancer suppressor gene promoter in tissues of oral leukoplakia on DNA level, risks of oral leukoplakia canceration can be assessed. If the promoter region for one or two of gene RASSF1A and gene MGMT shows a positive methylation result, a high risk of oral leukoplakia canceration can be predicted, while if the promoter region for gene RASSF1A and gene MGMT has a negative methylation result, a low risk of oral leukoplakia canceration can be predicted. The technical scheme disclosed in the invention can assist the diagnosis of a doctor, so that a clinician can better assess the canceration risk of leukoplakia patients based on the methylation level of oral leukoplakia patients and make an appropriate treatment scheme timely. Thus, the life quality of patients can be improved.

Description

【Technical field】 [0001] The invention relates to the field of epigenetics, more specifically, relates to the application of methylation detection of tumor suppressor gene promoters in the evaluation of oral leukoplakia carcinogenesis. 【Background technique】 [0002] At present, the judgment of cancer risk of oral leukoplakia mainly relies on biopsy. Pathologists judge the degree of epithelial cell proliferation in tissue sections according to the WHO international histological classification of tumors. Epithelial dysplasia is usually divided into three grades: mild, moderate and severe. When oral leukoplakia is accompanied by epithelial hyperplasia, its malignant potential increases with the degree of epithelial hyperplasia. Clinicians assess the cancer risk of oral leukoplakia according to whether there is abnormal hyperplasia and the degree of abnormal hyperplasia, carry out active relevant treatment, and follow up regularly, so as to play the role of early cancer preven...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 蒋伟文柳杨
Owner SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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