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Melanoma metastasis-associated marker and application thereof

A melanoma and genetic technology, applied in the field of disease diagnosis, can solve problems such as rising, diagnosis and treatment gaps

Pending Publication Date: 2019-10-11
BEIJING MEDINTELL BIOMED CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Epidemiological surveys also show that there are about 20,000 new cases in my country every year, while the mortality rate in Europe and the United States is basically stable. The mortality rate of MM is rising with the increase of the incidence rate, which shows that there is a gap in the diagnosis and treatment of MM in my country

Method used

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  • Melanoma metastasis-associated marker and application thereof
  • Melanoma metastasis-associated marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Screening for differentially expressed gene markers

[0034] 1. Sample collection

[0035] Primary group: primary lesion of melanoma, 3 cases

[0036] Metastatic group: melanoma metastatic lesions, the metastatic site is the lungs, 3 cases

[0037] 2. Tissue RNA total extraction

[0038] The tissue RNA extraction kit from TIANGEN Company was used to extract the total RNA. For details, please refer to the instruction manual.

[0039] 3. RNA quality detection

[0040] The purity and concentration of RNA were measured by Nanodrop2000 UV spectrophotometer, and the purity was higher when OD260 / OD280 was between 1.8-2.2. Agilent2100 measures the RIN value.

[0041] 4. High-throughput transcriptome sequencing

[0042] The samples were sequenced using Illumina Hiseq x-ten second-generation high-throughput sequencing technology.

[0043] The software cuffquant and cuffdiff were used for mRNA expression quantification and differential expression analysis. Cuffqu...

Embodiment 2

[0046] Example 2 QPCR verification screened differentially expressed gene markers

[0047] Based on the screening results of Example 1, DEGS2 or NKPD1 was selected for QPCR verification.

[0048] 1. Sample collection

[0049] According to the method of Example 1, 36 cases of melanoma primary lesion tissue and 33 cases of melanoma metastatic lesion tissue were collected.

[0050] 2. Extract tissue RNA

[0051] Step with reference to embodiment 1.

[0052] 3. Synthesize cDNA

[0053] First remove the genomic DNA reaction, add 5×gDNA Buffer 2.0μl, Total RNA 1μg, add RNase Free ddH to the test tube 2 O Bring the total volume to 10 μl, heat in a water bath at 42°C for 3 minutes, then add 2.0 μl of 10×Fast RT Buffer, 1.0 μl of RT Enzyme Mix, 2.0 μl of FQ-RT Primer Mix, RNase Free ddH 2 O 5.0 μl, after mixing, add to the above test tube and mix together a total of 20 μl, heat in a water bath at 42°C for 15min, then at 95°C for 3min.

[0054] 4. QPCR reaction

[0055] (1) Prime...

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Abstract

The invention discloses a melanoma metastasis-associated marker and application thereof. The associated marker is a DEGS2 gene or an NKPD1 gene. Expression of the DEGS2 gene or the NKPD1 gene is takenas an index for judging whether or not the melanoma metastasis phenomenon occurs. According to the melanoma metastasis-associated marker and the application thereof, a new diagnosis method is provided for clinical diagnosis for melanoma metastasis at the molecular level, and a product for diagnosing melanoma metastasis can be prepared and is suitable for clinical popularization.

Description

technical field [0001] The invention relates to the field of disease diagnosis, in particular to markers related to melanoma metastasis and applications thereof, and more specifically to a method for disease diagnosis by means of detecting abnormal expression of DEGS2 or NKPD1 genes. Background technique [0002] Melanoma, also known as malignant melanoma (MM), accounts for about 1% to 3% of all malignant tumors. Skin-derived melanoma, also known as cutaneous melanoma (cutaneous malignant melanoma, CMM), is derived from malignant transformation of melanocytes at the base of the epidermis. In Asians, the proportion of MM is about 50% to 70%. [0003] CMM accounts for about 3% of skin malignancies, but has a mortality rate as high as 65%. It is the most aggressive skin malignancy. It is more common in adults and the elderly, especially those over 40 years old, and less common in children; more men than women. The course of the disease is long. In the early stage, there are m...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886G01N33/574
CPCC12Q1/6886C12Q2600/158G01N33/5743G01N33/57484
Inventor 王麟唐美兰
Owner BEIJING MEDINTELL BIOMED CO LTD