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Application of an Epstein-Barr virus miRNA associated with nasopharyngeal carcinoma

A technology for Epstein-Barr virus and nasopharyngeal carcinoma, applied in the field of molecular biology, can solve problems such as limitations in early detection, inability to distinguish whether patients are infected or cured, and non-specific reactions have a large impact

Inactive Publication Date: 2011-12-07
SUN YAT SEN UNIV CANCER CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are still the following deficiencies: ①This test can only perform qualitative detection on samples, and cannot fully reflect the infection status of pathogens in vivo; The detection reagent cannot distinguish whether the patient is in the infection stage or has been cured, and cannot be used as an indicator to evaluate the prognosis of the disease and judge the effect of antiviral treatment; ③Because the EB virus infection window period is generally 2 to 4 weeks, early detection is subject to certain restrictions;④ Immunodeficiency patients have very low or no antibody levels, so the above methods cannot be used for detection;
Studies have found that EBV miRNA is abundant in nasopharyngeal carcinoma tissues. At present, there is no relevant report on the relationship between EBV miRNA and nasopharyngeal carcinoma, and there is no report on the application of EBV miRNA to the detection of nasopharyngeal carcinoma.

Method used

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  • Application of an Epstein-Barr virus miRNA associated with nasopharyngeal carcinoma
  • Application of an Epstein-Barr virus miRNA associated with nasopharyngeal carcinoma
  • Application of an Epstein-Barr virus miRNA associated with nasopharyngeal carcinoma

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Example 1: Detection of differentially expressed miRNAs between nasopharyngeal carcinoma tissues and corresponding adjacent normal tissues by miRNA chips

[0037] The present invention uses a micro-RNA (miRNA) chip (including 850 human and 39 EBV miRNAs, Exiqon product miRCURYLINA Array, Exiqon, Vedbaek, Denmark) to compare the miRNA expression differential profiles of five pairs of nasopharyngeal carcinoma samples. The analysis method of the chip is carried out according to the steps of the product instructions. The results showed that, compared with adjacent normal nasopharyngeal tissues, 60 miRNAs were expressed differently in nasopharyngeal carcinoma tissues, among which 50 miRNAs were upregulated and 10 miRNAs were downregulated. Among the 50 highly expressed miRNAs, 29 were encoded and transcribed by Epstein-Barr virus DNA. Table 1 lists the miRNAs with differential expression between nasopharyngeal carcinoma tissues and corresponding adjacent normal tissues dete...

Embodiment 2

[0042] Embodiment 2: A preferred embodiment of the miRNA differential expression profile experiment of EBV

[0043] In addition to the above-mentioned miRNA chips from Exiqon, 15 pairs of tissue samples were used. These samples were 20 pairs of nasopharyngeal carcinoma tissue samples obtained from the Royal Mary Hospital in Hong Kong in 2004. The tumors were taken separately with the help of fiberoptic nasopharyngoscope Nonneoplastic tissue with normal mucosa at the site of growth and corresponding adjacent carcinoma. The total RNA in the tissue was extracted and reverse-transcribed into cDNA by real-time fluorescent quantitative PCR to detect and verify 12 Epstein-Barr virus miRNAs (including BART6-5p, 6-3p, 1-3p, 5 , 17-5p, 8, 7, 14, 18-5p, 19-3p, 9 and 2-5p) in these 15 pairs of nasopharyngeal carcinoma and corresponding normal nasopharyngeal tissues.

[0044] Total RNA was extracted by using the mirVana microRNA extraction kit (Ambion, Austin, Tex) according to its operat...

Embodiment 3

[0048] Embodiment 3: A preferred example of comparing the expression levels of the above-mentioned 12 EBV miRNAs in 15 pairs of nasopharyngeal carcinoma samples and 2 EBV-positive cell lines

[0049] 50 up-regulated miRNAs were detected by miRNA chip, and 12 EBV miRNAs with high multiples of change, high abundance and possible target genes of tumor suppressor genes were selected, among which 5 came from the EBV BART1 region (BART6-5p , 6-3p, 1-3p, 5, 17-5p), 6 from the EBV BART2 region (BART8, 7, 14, 18-5p, 9, 19-3p) and BART2-5p. The copy numbers of EBV miRNAs in each cell of these 15 pairs of NPC samples were calculated to determine whether their expression levels were sufficient to down-regulate their target genes. The calculation method of the EBV miRNA copy number in each cell is as follows: Similar to the method used in Cosmopoulos et al, C666 was diluted to a serial known concentration, and then the Ct value of EBV miRNA at each dilution concentration was detected by qP...

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Abstract

The present invention provides an application of Epstein-Barr virus miRNA related to nasopharyngeal carcinoma, mainly the application of said Epstein-Barr virus miRNA in preparing a kit for diagnosis, prognosis judgment and treatment effect evaluation of nasopharyngeal carcinoma. The present invention also provides a kit for nasopharyngeal carcinoma diagnosis, prognosis judgment, and treatment effect evaluation, said kit comprising Epstein-Barr virus miRNA related to nasopharyngeal carcinoma. The Epstein-Barr virus miRNA related to nasopharyngeal carcinoma includes one or more sequences such as SEQ ID NO: 1-12. The 12 kinds of Epstein-Barr virus miRNAs provided by the present invention can be used for early diagnosis, prognosis judgment, treatment effect evaluation of nasopharyngeal cancer patients, and detection and follow-up of nasopharyngeal cancer high-risk groups by testing the expression level of Epstein-Barr virus miRNAs in serum. At the same time, the study of the target genes of these miRNAs can also provide evidence for Epstein-Barr virus tumorigenesis.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to the application of an Epstein-Barr virus miRNA related to nasopharyngeal carcinoma. Background technique [0002] Nasopharyngeal carcinoma is one of the common malignant tumors in southern China (including Guangdong, Guangxi and Hunan). The 5-year survival rate after radiotherapy for early nasopharyngeal carcinoma can reach 80-90%, while that for advanced nasopharyngeal carcinoma is only 20%. Therefore, strengthening the prevention, early detection, early diagnosis and early treatment of nasopharyngeal carcinoma is the key to improve the treatment effect of nasopharyngeal carcinoma. [0003] The occurrence and development of nasopharyngeal carcinoma is closely related to Epstein-Barr virus (EBV) infection. EBV belongs to the Y herpesvirus family. After the body is infected with EB virus, it can produce infectious mononucleosis, EB virus-associated hemophagocytic syndrome, X-link...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 关新元王雅欣邝丽云江家乐李焱
Owner SUN YAT SEN UNIV CANCER CENT
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