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Lung cancer diagnosis marker and application thereof

A lung cancer diagnosis and marker technology, applied in the field of biomedical detection, can solve the problem of not including the UHRF1 gene, and achieve the effect of high application value, high accuracy, and low requirements for instruments and equipment

Inactive Publication Date: 2018-03-09
天津脉络医学检验有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the gene detection products for lung cancer on the market involve more genes such as EGFR, KRAS, BRAF, ALK, MET, etc., basically do not contain UHRF1 gene, and there is no prior art that discloses that UHRF1 gene is related to lung cancer

Method used

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  • Lung cancer diagnosis marker and application thereof
  • Lung cancer diagnosis marker and application thereof
  • Lung cancer diagnosis marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Assembly of qRT-PCR detection kit

[0056] Specifically, the main reagents in the detection kit according to the present invention include:

[0057] PCR amplification reagents: 10× buffer, dNTPs, DNA polymerase, pure water, including PCR amplification primers for detecting UHRF1 gene loci and GAPDH internal reference gene primers, wherein,

[0058] The nucleotide sequence of the UHRF1 gene primer is shown in SEQ ID NO.1-2:

[0059] Upstream primer (SEQ ID NO.1): 5'-CCA GCA GAG CAG CCT CAT C-3';

[0060] Downstream primer (SEQ ID NO.2): 5'-TCC TTG AGT GAC GCC AGG A-3'.

[0061] The nucleotide sequence of the GAPDH internal reference gene primer is shown in SEQ ID NO.3-4:

[0062] Upstream primer (SEQ ID NO.3): 5'-CAG GAG GCA TTG CTG ATG AT-3';

[0063] Downstream primer (SEQ ID NO.4): 5'-GAA GGC TGG GGC TCA TTT-3'.

Embodiment 2

[0064] Example 2: Example of immunohistochemical detection of UHRF1

[0065] (1) Immunohistochemical detection: 1 normal tissue sample and 11 lung cancer patient tissue samples were extracted, embedded in paraffin and sectioned, and immunohistochemical experiments were performed using UHRF1 antibody. The specific experimental steps were as follows:

[0066] (a) Take a tissue sample with a size of 25px×25px×12.5px, slice it, put it in a constant temperature oven at 40°C, and dry it for later use;

[0067] (b) Dewaxing and hydration: bake in an oven at 65°C for 30 minutes, soak in xylene for 15 minutes×2, absolute ethanol for 5 minutes, 95% ethanol for 5 minutes, 85% ethanol for 5 minutes, PBS for 5 minutes×2, H2O for 1 minute;

[0068] (c) Antigen restoration: Place a high-temperature-resistant plastic slice rack in the microwave box, add antigen restoration solution, put the dewaxed and hydrated tissue slices after boiling, first heat for 5 seconds, then pause for 15 seconds, ...

Embodiment 3

[0080] Example 3: Example of immunohistochemical detection of UHRF1

[0081] (1) Immunohistochemical detection: 1 normal tissue sample and 4 tissue samples from patients with non-small cell lung cancer were extracted, embedded in paraffin and sectioned, and immunohistochemical experiments were performed using UHRF1 antibody;

[0082] The specific method of immunohistochemical detection is the same as that in Example 1.

[0083] (2) Analyze the expression of UHRF1 gene.

[0084] The results of immunohistochemical analysis were as follows: figure 2 shown, from figure 2 It can be seen that UHRF1 is not expressed in normal lung cells, but highly expressed in the nucleus of 59% of non-small cell lung cancer and adenocarcinoma.

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Abstract

The invention belongs to the technical field of biomedical detection and relates to a lung cancer diagnosis marker and application thereof. The invention discloses the lung cancer diagnosis marker; the lung cancer diagnosis marker is a UHRF1 gene. The biological marker has very high application value; a detection method is simple and the operation is simple and convenient; a new direction is provided for researches of a lung cancer diagnosis reagent and a theoretical foundation is provided for clinical pharmacy.

Description

technical field [0001] The invention belongs to the technical field of biomedical detection, and relates to a marker for lung cancer diagnosis and its application, in particular to a marker UHRF1 gene for lung cancer diagnosis and its application. Background technique [0002] Epidemiology points out that lung cancer is one of the malignant tumors with the fastest-growing morbidity and mortality. Among the causes of cancer-related deaths in the world, lung cancer ranks first. Non-small cell lung cancer (NSCLC) accounts for about 85% of all diagnosed primary lung cancers, and small cell lung cancer (SCLC) accounts for about 20%-25% of lung cancers. In recent years, scientific researchers have paid more and more attention to the study of tumor molecular biology, and found many tumor markers that can indicate the source and histological type of tumors. The detection of tumor markers has important clinical value in the screening, diagnosis, curative effect detection and prognos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11G01N33/574
CPCC12Q1/6886C12Q2600/136C12Q2600/158C12Q2600/166G01N33/57423G01N33/57484G01N2333/9015
Inventor 刘鹏飞张文盼王菁蕊
Owner 天津脉络医学检验有限公司
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