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Long-chain non-coding RNA marker for early diagnosis of malignant melanoma and application thereof

A malignant melanoma, long-chain non-coding technology, applied in the field of molecular diagnosis, can solve problems such as lack of new research, and achieve the effect of high sensitivity and specificity, high diagnostic characteristics, high clinical application potential and promotion effect

Pending Publication Date: 2020-05-01
THE FIRST AFFILIATED HOSPITAL OF MEDICAL COLLEGE OF XIAN JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is a lack of new research on its further mechanism

Method used

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  • Long-chain non-coding RNA marker for early diagnosis of malignant melanoma and application thereof
  • Long-chain non-coding RNA marker for early diagnosis of malignant melanoma and application thereof
  • Long-chain non-coding RNA marker for early diagnosis of malignant melanoma and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1qRT-PCR verifies the expression of Linc01296 in serum

[0038] 1. Experimental materials

[0039] 1. Serum collected from 98 healthy controls, skin benign moles, and malignant melanoma patients.

[0040] 2. Dye-based Linc01296 qRT-PCR kit, the components are as follows:

[0041] (1) Linc01296 upstream primer (sequence such as SEQ ID NO.2)

[0042] (2) Linc01296 downstream primer (sequence such as SEQ ID NO.3)

[0043] (3) U6 upstream primer (internal reference calibration gene, sequence such as SEQ ID NO.4)

[0044] (4) U6 downstream primer (internal reference calibration gene, sequence such as SEQ ID NO.5)

[0045] (5) 2×one-step reverse transcription-fluorescent quantitative PCR mixture

[0046] (6) Hot start Taq enzyme

[0047] (7) RTase reverse transcriptase

[0048] (8) DEPC-ddH 2 o

[0049] 2. Experimental method

[0050] 1. Total RNA Extraction from Serum Samples

[0051] The total RNA in the serum of healthy controls, skin benign moles and...

Embodiment 2

[0085] Example 2 ROC curve analysis of the diagnostic value of serum Linc01296 expression in malignant melanoma

[0086] On the basis of detecting the relative expression levels of Linc01296 in the serum of malignant melanoma patients and healthy controls, the ROC curve was drawn using SPSS 22.0 software for the qRT-PCR reaction results. The result is as image 3 It shows that the area under the ROC curve of serum Linc01296 for the diagnosis of malignant melanoma is 0.896 (95% CI: 0.847-0.946; P<0.001), the optimal diagnostic threshold of serum Linc01296 for colorectal cancer is 2.149, and the sensitivity is 84.69%. The specificity was 88.77%. Serum samples from patients with malignant melanoma positive for Linc01296 were selected according to the diagnostic threshold of 2.149 determined in this study. Then, the dye-based fluorescent quantitative kit of the present invention was used to carry out repeated experiments on the positive samples for 3 consecutive times, and the r...

Embodiment 3

[0088] Example 3 qRT-PCR detection of expression of Linc01296 in serum of patients with malignant melanoma before and after radical operation

[0089] 1. Experimental materials

[0090] 1. Serum collected from 76 patients with malignant melanoma who underwent radical resection before and 3 months after surgery.

[0091] 2. Dye-based Linc01296 qRT-PCR kit (the components are the same as above).

[0092] 2. Experimental method

[0093] 1. Total RNA Extraction from Serum Samples

[0094] The total RNA in the serum of healthy controls, skin benign moles and malignant melanoma patients was extracted according to the reagents and steps required by Tiange Biochemical Technology (Beijing) Co., Ltd. High Efficiency Blood Total RNA Extraction Kit (Product No. DP443). The specific steps are as mentioned above.

[0095] 2. Amplification of Linc01296: According to the reagents and steps required by the one-step reverse transcription-fluorescence quantitative kit (product number FP303) ...

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Abstract

The invention relates to a long-chain non-coding RNA marker for early diagnosis of malignant melanoma and application thereof. The long-chain non-coding RNA marker is Linc01296 with a corresponding DNA sequence as shown in SEQ. ID. NO. 1. The application is an application of the long-chain non-coding RNA marker Linc01296 in the preparation of an early diagnosis reagent for malignant melanoma as atarget RNA. According to the invention, lcnRNA, Linc0126, with abnormal expression in serum of malignant melanoma is successfully screened, and expression of Linc01296 is significantly high in the serum of malignant melanoma patients. In addition, Linc01296 can effectively distinguish malignant melanoma patients from healthy people and patients with benign skin moles. The marker has high sensitivity and specificity, wherein the sensitivity can reach 84.69%, the specificity can reach 88.77%, and the marker has high diagnostic characteristics.

Description

technical field [0001] The invention belongs to the technical field of molecular diagnosis, and relates to a long-chain non-coding RNA and its application, in particular to a long-chain non-coding RNA marker which can be used for the early diagnosis of malignant melanoma and its application. Background technique [0002] Malignant melanoma originates from neural crest melanocytes and is the most lethal tumor among surface malignant tumors. It has become one of the fastest-growing malignant tumors in my country. Early detection of malignant melanoma can be cured by surgery. However, it is highly invasive, develops rapidly, is insensitive to radiotherapy and chemotherapy, and some patients have metastasized when they see a doctor. These patients have poor prognosis and high mortality (Siegel RL, Miller K D, Jemal A. Cancer statistics, 2018[J ]. CA Cancer JClin, 2018, 68(1): 7-30.). Therefore, early diagnosis and early treatment of malignant melanoma are extremely important f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/113C12Q1/6851
CPCC12Q1/6886C12Q1/6851C12Q2600/158C12Q2561/113C12Q2563/107C12Q2531/113C12Q2521/107
Inventor 王丽娟史圣甲穆欣葛睿杨莹莹
Owner THE FIRST AFFILIATED HOSPITAL OF MEDICAL COLLEGE OF XIAN JIAOTONG UNIV
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