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Screening reagent for detecting core molecules of DNA damage signal path and application of screening reagent

A DNA damage and signaling pathway technology, applied in the field of molecular biology, can solve problems such as premature aging, increased cancer tendency, immune deficiency, etc.

Pending Publication Date: 2020-10-30
THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Inherited DNA repair defects can be catastrophic, leading to developmental abnormalities, increased predisposition to cancer, immune deficiencies, neurodegenerative diseases and premature aging

Method used

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  • Screening reagent for detecting core molecules of DNA damage signal path and application of screening reagent
  • Screening reagent for detecting core molecules of DNA damage signal path and application of screening reagent
  • Screening reagent for detecting core molecules of DNA damage signal path and application of screening reagent

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Embodiment 1

[0294] The present invention is illustrated by human liver cancer Huh7 and liver cancer cell Huh7 treated with SR-4835, but the present invention is not limited to these two types of cells, and the detection reagent of the present invention can be applied to liver cancer cell lines including SMMC7721, HCCLM3, etc. It can detect DNA damage signals in clinical liver cancer tissues and paracancerous tissues such as lung cancer, gastric cancer, pancreatic cancer, ovarian cancer and other tumors.

[0295] In the present invention, the human liver cancer cell line Huh7 is cultivated, and then treated with SR-4835 (100nM) for 24 hours, and the change of core molecules in the treated cell line is detected by the screening reagent of the DNA damage signaling pathway of the present invention.

[0296] The human liver cancer Huh7 used in the present invention was purchased from ATCC Company of the United States, the Shanghai Cell Bank, DMEM medium, and fetal bovine serum were purchased fr...

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Abstract

The invention provides a screening reagent for detecting core molecules of DNA damage signal path and an application of the screening reagent. The PCR reagent comprises PCR reaction primers for detecting ABL1 genes, APEX1 genes, ATM genes, ATR genes and other related genes, reference genes and the like. According to the invention, core molecules related to a DNA damage signal path are concentratedon a flat plate; a real-time fluorescent quantitative PCR reaction is carried out once; core molecules for regulating and controlling DNA damage signal pathways in cells are reflected, a drug-treatedliver cancer Huh7 and an untreated liver cancer Huh7 are compared for example, possible pathways of the core signal molecules causing DNA damage are discussed, and most direct evidences are providedfor researching regulation and control of key proteins; according to the invention, the core signal molecule of DNA damage is rapidly and accurately found from the transcription level, and a powerfultool is provided for mechanism discussion of new targeting drugs, development of targeting tumor inhibitors and the like.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a screening reagent for detecting core molecules of a DNA damage signaling pathway and an application thereof. Background technique [0002] Every cell of our body produces thousands of DNA lesions every day due to various internal and environmental factors. These lesions, if not properly repaired, can lead to mutations and broader genomic aberrations that threaten cell viability and ultimately lead to cancer. DNA lesions will trigger a series of well-organized and multifaceted synergistic responses inside the cell to jointly combat this genotoxic stress, to detect DNA damage, indicate its existence, and mediate its repair. This series of defense responses That is called DNA damage response (DNA damage response, DDR). DNA repair is critical in many biological processes, including the development of the immune system, meiotic recombination, and the prevention of a va...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6851C12N15/11
CPCC12Q1/6886C12Q1/6851C12Q2600/158C12Q2600/136C12Q2561/113C12Q2563/107C12Q2545/101C12Q2531/113
Inventor 刘世海贺桂芳潘华政刘畅畅蔡铎
Owner THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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