Reagent for detecting Notch signal path as well as PCR (Polymerase Chain Reaction) detecting method and application thereof

A technology of signaling pathways and reagents, applied in the field of molecular biology

Inactive Publication Date: 2014-03-12
HOSPITAL ATTACHED TO QINGDAO UNIV
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Problems solved by technology

[0005] Although Notch signaling plays an important role in the process of the origin of life, and also exhibits tumor suppressor properties in a small number of tumors, abnormal

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  • Reagent for detecting Notch signal path as well as PCR (Polymerase Chain Reaction) detecting method and application thereof
  • Reagent for detecting Notch signal path as well as PCR (Polymerase Chain Reaction) detecting method and application thereof
  • Reagent for detecting Notch signal path as well as PCR (Polymerase Chain Reaction) detecting method and application thereof

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Embodiment 1

[0285] The present invention is illustrated by breast cancer cells, but the present invention is not limited to breast cancer, and the detection reagent of the present invention can be applied to common cancers including breast cancer, lung cancer, liver cancer and the like.

[0286] The invention cultivates two cell lines: human breast cancer cell line HS578T and human breast normal cell line HS578BST, and then detects changes of core NOTCH molecules in the two cell lines by using a NOTCH signal path detection reagent.

[0287] The human breast cancer cell line HS578T and human breast normal cell line HS578BST used in the present invention were purchased from ATCC Company of the United States, DMEM medium and fetal bovine serum were purchased from Gibco Company of the United States, RNAiso for extracting total RNA, reverse transcription kit, Premix Ex Taq kit was purchased from TaKaRa.

[0288] Using the reagent of the present invention to detect the intracellular NOTCH sign...

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Abstract

The invention provides a reagent for detecting a Notch signal path as well as a PCR (Polymerase Chain Reaction) detecting method and application thereof. The detecting reagent comprises PCR reaction primers for detecting an ADAM10 gene, an ADAM17 gene, an AES gene, a CBL gene, a CCND1gene, a CD44 gene, and the like, and primers of corresponding internal-reference regulatory genes GAPDH (Reduced Glyceraldehyde-Phosphate Dehydrogenase), ACTB (Beta-Actin) and B2M (Beta-2-Microglobulin). The invention provides the reagent for detecting core molecule changes of the NOTCH signal channel; by virtue of the detecting reagent, genes associated with the NOTCH channel can be quickly detected on transcriptional level by virtue of real-time fluorescent quantitative PCR; on this basis, core molecules and a mechanism of action of a tumor NOTCH signal channel are analyzed and researched, so that an NOTCH regulatory channel of tumor-associated medicaments is quickly and accurately found, and therefore, a powerful tool is provided for screening anti-tumor medicaments, discussing the mechanism of a novel targeted medicament, and the like.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a reagent for detecting Notch signaling pathway, a PCR detection method and an application thereof. Background technique [0002] Notch signaling is a highly conserved signal pathway in genetic evolution that reflects the communication between adjacent cells. related to development. [0003] The Notch signaling pathway is composed of transmembrane receptors, ligands, nuclear binding proteins and target genes. Notch receptors include Notch extracellular domains and transmembrane regions / Notch intracellular domains. They are cell surface receptors and nuclear receptors. A single-chain transmembrane protein with dual functions of internal transcriptional regulation. Currently, there are four types of Notch receptors found in mammals, namely Notch-1, Notch-2, Notch-3 and Notch-4, which are usually activated by the mutual triggering of Notch ligands in adjacent cells. N...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/6886
Inventor 王海波梁晔刘相萍王丽萍刘世海姚如永隋爱华杨堃迟静薇周泉刘加秀
Owner HOSPITAL ATTACHED TO QINGDAO UNIV
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