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Serum/plasma miRNA serum marker related to cervical carcinoma and precancerous lesions thereof and application thereof

A technology of precancerous lesions and markers, applied in the fields of genetic engineering and oncology

Active Publication Date: 2013-05-29
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] However, there is no report on the use of serum / plasma miRNAs in the auxiliary early diagnosis of cervical cancer. If the serum / plasma miRNAs abnormally expressed in cervical cancer and its precancerous lesions can be screened as biomarkers, and the corresponding disease Progress monitoring and auxiliary diagnostic kits will definitely be a powerful impetus to the diagnosis status of cervical cancer in my country

Method used

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  • Serum/plasma miRNA serum marker related to cervical carcinoma and precancerous lesions thereof and application thereof
  • Serum/plasma miRNA serum marker related to cervical carcinoma and precancerous lesions thereof and application thereof
  • Serum/plasma miRNA serum marker related to cervical carcinoma and precancerous lesions thereof and application thereof

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Embodiment 1

[0080] The collection of embodiment 1 sample and the arrangement of sample data

[0081] The inventor has collected a large number of peripheral blood samples of cervical cancer patients and controls from the First Affiliated Hospital of Nanjing Medical University, Nantong Cancer Hospital and Nanjing Jiangning Community since March 2006. By sorting out the sample data, the inventor selected 105 samples meeting the following criteria were selected as experimental samples for Solexa sequencing and subsequent series of qRT-PCR verification:

[0082] 1. New cases of cervical cancer;

[0083] 2. No surgery, radiotherapy and chemotherapy before blood collection, no preoperative radiotherapy and chemotherapy;

[0084] 3. Healthy female controls matched with the age of the case;

[0085] 4. Age-matched CIN stage I / II / III cases.

[0086] The demographic and clinical data of these samples were collected systematically.

Embodiment 2

[0087] Example 2 Solexa sequencing experiment of miRNA in serum / plasma

[0088] Among the 30 eligible cervical cancer patients and 30 healthy female controls mentioned above, the two groups were age-matched. The two groups of people were subjected to Solexa sequencing test to obtain relevant results. The specific steps are:

[0089] 1. Take 50ml of serum from patients in the "cervical cancer case" group and the "healthy female control" group, and add an equal volume of Trizol reagent;

[0090] 2. Phase separation: place at room temperature for 15 minutes, then add chloroform according to the volume ratio of 0.2ml chloroform / 1ml Trizol reagent, shake for 15s, room temperature for 15 minutes, centrifuge at 12,000g, 4°C for 15 minutes;

[0091] 3. Transfer the aqueous phase to a new 50ml centrifuge tube, and remove the protein phase in 3 steps of phenol / chloroform;

[0092] 4. RNA precipitation: transfer the aqueous phase to a new centrifuge tube, add isopropanol according to ...

Embodiment 3

[0101] qRT-PCR experiment of miRNA in embodiment 3 serum / plasma

[0102] According to the above-mentioned Solexa results, miRNAs that meet the following conditions are selected for further verification by qRT-PCR method: 1) miRNAs with a fold difference of 5 times in the two groups are used as serum biomarkers for preliminary screening in the present invention, 2) these miRNAs The copy number in at least one group ("cervical cancer case" group and "healthy female control" group) is greater than 50 to improve detection efficiency. miRNAs that meet the above conditions include: miR-1, miR-21, miR-23b, miR-28-3p, miR-29a, miR-100, miR-122, miR-125b, miR-181a, miR-206, miR -483-5p. Primers for reverse transcription and qRT-PCR were designed according to the selected miRNAs (see Table 4). The qRT-PCR detection of miRNA was performed on the serum individual individuals of the "cervical cancer case" group and the "healthy female control" group. Strict quality control was implement...

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Abstract

The invention belongs to the fields of gene engineering and oncology, and discloses a serum / plasma miRNA serum marker related to cervical carcinoma and precancerous lesions thereof and application thereof. The marker is single miR-21 or the combination of miR-21 and miR-29a. The marker and the primer thereof can be used for preparing diagnosis kits which are used for assistant early diagnosis of cervical carcinoma and precancerous lesions thereof.

Description

technical field [0001] The invention belongs to the fields of genetic engineering and oncology, and relates to a serum / plasma miRNA marker related to cervical cancer and its precancerous lesions and an application thereof. Background technique [0002] Cervical cancer is one of the most common malignant tumors in women, accounting for the second place in the world's female malignant tumors, second only to breast cancer. In 2002, there were 490,000 new cases of cervical cancer in the world, 83% of which occurred in developing countries, and 270,000 people died of cervical cancer. Although the mortality rate of cervical cancer in my country's cities has shown a downward trend since the 1960s, the incidence of cervical cancer and its mortality rate in rural areas have remained high. Especially in the late 1990s, the incidence of cervical cancer showed an obvious trend of younger people, and the incidence of cervical cancer in urban women aged 35-44 was increasing at a rate of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 胡志斌沈洪兵李瑛张辰宇董静
Owner NANJING MEDICAL UNIV
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