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Malignant melanoma biomarker PTGER2 and application thereof

A melanoma and drug technology, applied in the field of biomedicine, can solve problems such as the current situation is not optimistic, and there is no significant progress in clinical treatment

Pending Publication Date: 2021-04-20
河北仁博科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no significant progress in clinical treatment, and the current situation is still not optimistic

Method used

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  • Malignant melanoma biomarker PTGER2 and application thereof
  • Malignant melanoma biomarker PTGER2 and application thereof
  • Malignant melanoma biomarker PTGER2 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The mRNA levels of PTGER2 in mouse normal epidermal cell JB6, mouse melanoma high-metastatic cell B16-F10 and mouse melanoma low-metastatic cell B16-F1 were compared.

[0028] Mouse melanoma cells B16-F10 were inoculated in DMEM medium containing 10% fetal bovine serum (adding penicillin and streptomycin 100 U / mL), placed at 37°C, 5% CO 2 Cultured in an incubator, digested and passaged with 0.25% trypsin.

[0029] The operation steps are as follows:

[0030] RNA extraction and fluorescent quantitative PCR detection: Use the total RNA rapid extraction kit (Flying Biotech) to extract the RNA of the cells, and perform reverse transcription according to the conventional steps to form cDNA, and then detect the expression effect of PTGER2 by real-time fluorescent quantitative PCR. Among them, The sequences of cDNA amplification primer pairs are:

[0031] qPCR primer F: 5'-CAGCTCGGTGATGTTCTCGG-3' (SEQ ID NO: 1);

[0032] qPCR primer R: 5'-GAGCACCAATTCCGTTACCAG-3' (SEQ ID NO...

Embodiment 2

[0035] The protein levels of PTGER2 in mouse normal epidermal cells and mouse melanoma high-metastatic cells B16-F10 and mouse melanoma low-metastatic cells B16-F1 were compared.

[0036] The operation steps are as follows:

[0037] Collect the cells at 80% confluence, discard the supernatant after centrifugation, rinse twice with PBS, and discard the supernatant. Add RIPA lysate and lyse on ice for 20 min. The supernatant was collected by centrifugation at 12000 g for 10 min. Add 1xSDS sample buffer, mix well by pipetting and then boil for denaturation for 5min. Total proteins were separated by 10% SDS-PAGE gel and then transferred to PVDF membrane. 5% BSA was blocked at room temperature for 2 hours, incubated with PTGER2 antibody (GeneTex, GTX66706 (1:1000 dilution)) overnight at 4°C, and washed 3 times with TBST. The secondary antibody was incubated at room temperature for 1 h, and washed 3 times with TBST. ECL ultra-sensitive chemiluminescent solution was developed an...

Embodiment 3

[0040] The expression levels of PTGER2 in human normal epidermal cells HaCat and human melanoma cells SK-MEL-24 and SK-MEL-28 were compared.

[0041] For specific steps, refer to the experimental method described in Example 1.

[0042] The sequences of cDNA amplification primer pairs are:

[0043] qPCR primer F: 5'-CGATGCTCATGCTCTTCGC-3' (SEQ ID NO: 3);

[0044] qPCR primer R: 5'-GGGAGACTGCATAGATGACAGG-3' (SEQ ID NO: 4).

[0045] The result is as image 3 As shown, the expression level of PTGER2 was found to be lower in human melanoma cells than in normal epidermal cells.

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Abstract

The invention discloses application of a reagent for detecting a PTGER2 gene or an expression product of the PTGER2 gene in preparation of a product for diagnosing malignant melanoma. The invention proves that the PTGER2 is low in expression in malignant melanoma cells of mice and human beings and can be used as a malignant melanoma diagnosis or prognosis marker. Furthermore, the statistical correlation between the expression level of the PTGER2 and malignant melanoma diseases is analyzed through 451 malignant melanoma samples in a TCGA database, and it is found that the lifetime of a patient with the high expression level of the PTGER2 is obviously superior to that of a patient with the low expression level of the PTGER2. The correlation between the PTGER2 and the malignant melanoma is proved for the first time, and the PTGER2 is a very valuable potential drug screening or treatment target in the field of malignant melanoma treatment.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a melanoma biomarker PTGER2 and its use. Background technique [0002] Melanoma, also called malignant melanoma (MM), accounts for about 1% to 3% of all malignant tumors. Cutaneous melanoma (CMM) is an aggressive tumor derived from the malignant transformation of epidermal melanocytes. It is the malignant tumor with the fastest progression and the poorest prognosis among all skin tumors, with blood supply or lymph node metastasis in the later stage. Its onset is insidious, and it is prone to early metastasis. Most of the patients who seek medical treatment are in the middle and late stages, and the mortality rate within 5 years of metastasis is high. In recent years, the morbidity and mortality of MM have shown an obvious upward trend. The American Cancer Society (ACS) reported in 2017 that, except for the morbidity and mortality of MM and thyroid cancer, which were on the r...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886G01N33/68G01N33/574A61K45/00A61P35/00
Inventor 崔健姜薇
Owner 河北仁博科技有限公司
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