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Biological marker for evaluating EB virus-positive nasopharyngeal carcinoma DNA (Deoxyribonucleic Acid) damage repair capacity and radiation therapy prognosis and application thereof

A biomarker, DNA damage technology, applied in the field of biomarkers

Active Publication Date: 2016-08-17
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Epstein-Barr virus infection-related tumors account for about 1.5% of global malignant tumors, accounting for 5.6% of all infection-related tumors, and there is currently no EBV vaccine

Method used

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  • Biological marker for evaluating EB virus-positive nasopharyngeal carcinoma DNA (Deoxyribonucleic Acid) damage repair capacity and radiation therapy prognosis and application thereof
  • Biological marker for evaluating EB virus-positive nasopharyngeal carcinoma DNA (Deoxyribonucleic Acid) damage repair capacity and radiation therapy prognosis and application thereof
  • Biological marker for evaluating EB virus-positive nasopharyngeal carcinoma DNA (Deoxyribonucleic Acid) damage repair capacity and radiation therapy prognosis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 EBV-LMP1 inhibits DNA double-strand break repair in nasopharyngeal carcinoma cells

[0024] H2AX protein is a member of the H2A histone family in eukaryotic cells, and its carboxy-terminus includes a serine-glutamine-glutamic acid (Ser-Gin-Glu, SQE) domain with a serine residue at position 139, which can be The PI3K kinase family includes ATM, ATR, DNA-PK phosphorylation, namely γ-H2AX. Phosphorylation of H2AX Ser139 is an early event of DNA double-strand breaks caused by ionizing radiation or chemical drugs. γ-H2AX foci are formed at the breaks to recruit DNA damage recognition and repair proteins. Currently, γ-H2AX is considered to be one of the most sensitive and specific indicators for detecting DNA double-strand breaks. The effect of EBV-LMP1 on the repair of DNA double-strand breaks in nasopharyngeal carcinoma cells was clarified by Western Blot detection of γ-H2AX.

[0025] The experimental method is as follows: Nasopharyngeal carcinoma cells were cul...

Embodiment 2

[0027] Example 2 EBV-LMP1 inhibits DNA-PK phosphorylation in nasopharyngeal carcinoma cells

[0028]The phosphorylation of DNA-PK Thr2609 is the key to its role in the repair of DNA double-strand breaks. Western Blot detection of DNA-PK Thr2609 in nasopharyngeal carcinoma cells stably expressing LMP1 and knocking out LMP1 confirmed that EBV-LMP1 has a role in nasal Regulation of DNA-PK phosphorylation in pharyngeal carcinoma cells.

[0029] The experimental method is as follows: nasopharyngeal carcinoma cells were cultured in a 6-well plate, EBV-LMP1 positive cells were transfected with LMP1 siRNA and cultured for 48 hours, then 4Gy was irradiated with 6-MV X-rays, and the cells were extracted 0.5 hours after irradiation The total protein was analyzed by Western Blot of DNA-PK Thr2609.

[0030] The results showed that EBV-LMP1 could inhibit the phosphorylation of DNA-PK Thr2609 in the two strains of EBV-LMP1 positive cells, and the phosphorylation of DNA-PK Thr2609 was promot...

Embodiment 3

[0031] Example 3 EBV-LMP1 inhibits the combination of DNA-PK and AMPK in nasopharyngeal carcinoma cells and reduces the phosphorylation of AMPK

[0032] The physical combination of DNA-PK and AMPK induced by DNA damage and the regulation of this combination by EBV-LMP1 were detected by co-immunoprecipitation, and the phosphorylation of Thr172 of AMPK induced by DNA damage was detected by Western Blot.

[0033] The experimental method is as follows: nasopharyngeal carcinoma cells were cultured in a 10cm culture dish. When the cells reached 90% confluence, 4Gy of 6-MV X-rays were irradiated, and the total protein of the cells was extracted 0.5h after the irradiation. Remove non-specific proteins that can bind to beads in the whole protein lysate: ① Take 20ul of A-sepharose bead suspension, centrifuge to remove the supernatant, and wash three times with ice-cold 1×PBS. ② Prepare 1000ug of whole protein lysate for each experimental group, add 20ul of washed beads, shake at 4°C for...

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Abstract

The invention discloses a biological marker for evaluating EB virus-positive nasopharyngeal carcinoma DNA (Deoxyribonucleic Acid) damage repair capacity and radiation therapy prognosis and application thereof. The biological marker is phosphorylated DNA-PK (Protein Kinase) and AMPK (Adenosine Monophosphate Protein Kinase), preferably phosphorylated DNA-PK Thr2609 and AMPK Thr172. The biological marker can be used for preparing a detection drug and a kit for evaluating the EB virus-positive nasopharyngeal carcinoma DNA damage repair capacity and the radiation therapy prognosis.

Description

technical field [0001] The present invention relates to biomarkers, specifically, the phosphorylation levels of DNA protein kinase (DNA-activated protein kinase, DNA-PK) and AMP-activated protein kinase (AMP-activated protein kinase, AMPK) involved in DNA damage response are used to evaluate Epstein-Barr virus (Epstein–Barr virus, EBV) positive nasopharyngeal carcinoma DNA damage repair ability and radiotherapy prognosis biomarkers. Background technique [0002] DNA damage response (DNA damage response, DDR) detects and transmits DNA damage signals through multiple signal transduction pathways, initiates cell cycle checkpoints, repairs DNA damage or activates apoptosis, maintains genome stability and cell homeostasis . Deficiency in cellular DNA damage repair ability is an important mechanism of tumor pathogenesis. Unrepaired or wrongly repaired DNA damage may cause genome instability and cause tumors; at the same time, the abnormality of DNA damage repair is related to th...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/118C12Q2600/158
Inventor 曹亚卢景琛
Owner CENT SOUTH UNIV
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