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Application of miR-17, miR-20a, miR-29c and miR-223 as nasopharyngeal carcinoma molecular markers

A technology of mir-20a and mir-223, applied in the application field of microRNA in the screening and early diagnosis of high-risk groups of nasopharyngeal carcinoma, to achieve the effect of non-invasive diagnosis

Inactive Publication Date: 2014-12-24
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, the expression levels of serum and plasma microRNAs in different healthy individuals are quite consistent without significant individual differences, while tumors can significantly affect the expression of microRNAs in serum and plasma

Method used

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  • Application of miR-17, miR-20a, miR-29c and miR-223 as nasopharyngeal carcinoma molecular markers
  • Application of miR-17, miR-20a, miR-29c and miR-223 as nasopharyngeal carcinoma molecular markers
  • Application of miR-17, miR-20a, miR-29c and miR-223 as nasopharyngeal carcinoma molecular markers

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Experimental program
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Effect test

Embodiment 1

[0013] Example 1 Detection of miR-17, miR-20a, miR-29c and miR-223 in peripheral blood serum samples

[0014] 1. Serum collection

[0015] (1) Use a BD5ml serum collection tube to collect venous fasting blood, fill the tube as much as possible, about 4ml, and gently invert it 3 times;

[0016] (2) Stand at room temperature for 1 hour;

[0017] (3) Centrifuge, ×1600g, 10min, 4°C;

[0018] (4) Carefully pipette the upper layer of serum on ice to 1.5ml Tube, about 600μl / tube, number and label, and store at -80°C;

[0019] 2. Extraction of serum total RNA

[0020] (1) Thaw serum samples on ice

[0021] (2) Take a serum sample (500 μl / tube) and add an equal volume of 2× denaturing solution, vortex and mix, and incubate on ice for 5 minutes

[0022] (3) Add an equal volume (1000μl / tube) of acidic phenol: chloroform, shake for 60sec

[0023] (4) Centrifuge (12000g, room temperature, 5min), absorb the upper aqueous phase to a new Tube

[0024] (5) Repeat steps 2 and 3 twice

...

Embodiment 2

[0065] Example 2 Clinical verification of miR-17, miR-20a, miR-29c and miR-223 discriminant equations in serum of normal people and nasopharyngeal carcinoma patients

[0066] According to the above method, the expression of 4 miRNAs in the serum of 57 normal people and 74 patients with nasopharyngeal carcinoma was detected. Compared with the A value in normal human serum, the expressions of miR-17, miR-20a, miR-29c and miR-223 in the peripheral blood of 72 patients were significantly different, and the A>-3.30 was positive, indicating nasopharyngeal carcinoma Risk positive, and confirmed by histopathological examination are all patients with nasopharyngeal carcinoma, the calculated serum A value of 55 normal subjects were all <-3.30, the specificity was 96.5%, and the sensitivity was 97.3%.

[0067] The above studies have shown that the four miRNAs in peripheral blood serum can be used as molecular markers for the diagnosis of nasopharyngeal carcinoma patients. When the discri...

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Abstract

The invention discloses an application of miR-17, miR-20a, miR-29c and miR-223 as nasopharyngeal carcinoma molecular markers. The invention provides the 4 micro-RNAs in serum and as the nasopharyngeal carcinoma molecular markers and a detection method thereof, wherein the detection method comprises the steps: total RNAs in peripheral blood serum are extracted and subjected to reverse transcription into cDNAs, Ct values of the 4 micro-RNAs are obtained through a real-time quantitative PCR method, and the A value is calculated through a formula of A=(Ct[miR-29c]+Ct[miR-223])-(Ct[miR-17]+Ct[miR-20a]); if the A value is greater than -3.30, the result is positive, and a patient is prompted to be suffered from nasopharyngeal carcinoma; and if the A value is less than -3.30, the result is negative, and the patient is prompted to be not suffered from nasopharyngeal carcinoma. Through detection of expression conditions of the 4 micro-RNAs in the peripheral blood of various crowds, the risk of attacking nasopharyngeal carcinoma is predicted, the detection method is used for screening crowds with high risk of nasopharyngeal carcinoma, and the rapid and noninvasive diagnosis on nasopharyngeal carcinoma patients is achieved.

Description

technical field [0001] The invention relates to tumor molecular markers, in particular to the application of microRNA in the screening and early diagnosis of nasopharyngeal carcinoma high-risk groups. technical background [0002] Nasopharyngeal carcinoma has obvious geographical and ethnic distribution characteristics. my country is a high-incidence area of ​​nasopharyngeal carcinoma, especially in southern China. According to statistics, the incidence of male nasopharyngeal carcinoma is 15-50 / 100,000, ranking third among malignant tumors. Taiwan is also a high-incidence area for nasopharyngeal carcinoma, with an incidence rate of 8.3 / 100,000 and 3.5 / 100,000 for men and women respectively. In Western countries such as Europe and the United States, nasopharyngeal carcinoma is rare, and nasopharyngeal carcinoma is also vividly called "China's cancer". The onset of nasopharyngeal carcinoma is hidden, there is no obvious clinical manifestation in the early stage, it is not easy ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/113
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 李桂源彭淑平曾希李夏雨牛蔓韦娉嫔
Owner CENT SOUTH UNIV
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