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Marker for diagnosing liver cancer, and detection reagent and application of marker

A marker, liver cancer technology, applied in the field of biomedicine, can solve problems such as normal physiological regulation disorder and tumor occurrence

Inactive Publication Date: 2020-09-04
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The currently known signaling pathways involved in the occurrence and development of primary liver cancer mainly include NF-κB pathway, autophagy pathway, PI3K / AKT / mTOR signaling pathway and other signaling pathways. These gene mutations and related signaling pathways related to the occurrence and development of liver cancer Alteration affects biological processes such as cell proliferation, apoptosis, and cell differentiation, leading to the disorder of its normal physiological regulation and eventually leading to tumorigenesis

Method used

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  • Marker for diagnosing liver cancer, and detection reagent and application of marker
  • Marker for diagnosing liver cancer, and detection reagent and application of marker
  • Marker for diagnosing liver cancer, and detection reagent and application of marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1 cell line and culture condition

[0026] Human embryonic kidney epithelial cells 293, human liver cancer cell lines HepG2, Huh7, Hep3B, PLC / PRF / 5, SK-HEP-1, human normal liver cells HL-7702 cells, long-term cultured cell lines in our laboratory, culture conditions : All adopt DMEM medium (containing 1% penicillin and streptomycin), place 5% CO2, and cultivate in a 37°C incubator.

Embodiment 2

[0027] Example 2 Detection of TRAF2 mRNA expression levels in liver cancer cells and normal liver cells by Real-time PCR

[0028] 1. RNA extraction

[0029] (1) Take the cells cultured in the six-well plate, add 1ml Trizol to lyse the cells, and transfer the liquid to an RNase-free Eppendorf tube.

[0030] (2) Add 0.2ml of pre-cooled chloroform, shake vigorously by hand for 15 seconds, and then let it stand at room temperature for 3 minutes until a clear layering phenomenon can be seen.

[0031] (3) Centrifuge at 12,000g for 15 minutes at 4°C, and transfer the upper RNA-containing aqueous phase to a new Eppendorf tube.

[0032] (4) Add 0.5ml of pre-cooled isopropanol, mix upside down, and let stand at room temperature for 10 minutes.

[0033] (5) Centrifuge at 10,000 g for 10 minutes at 4°C; discard the supernatant, and add 1 ml of 75% ethanol to wash the RNA pellet. Centrifuge at 7500g for 5 minutes at 4°C and discard the supernatant.

[0034] (6) Dry at room temperature ...

Embodiment 3

[0046] Example 3 Detection of TRAF2 protein expression levels in cancer tissues and paracancerous tissues in clinical specimens from patients with liver cancer.

[0047] The present invention has a total of 120 pairs of clinical tissue samples, of which 90 pairs of tissue chips are used for immunohistochemical staining to detect the expression level of TRAF2, and 30 pairs are used to detect the protein expression level of TRAF2 by Western blot. The tissues are from the First Affiliated Hospital of Zhejiang University.

[0048] 1. Extraction of histone

[0049] (1) Grinding with liquid nitrogen, placing the obtained liver cancer clinical sample tissue in a mortar, adding liquid nitrogen for grinding, and ensuring that there is always liquid nitrogen in the mortar during the grinding process. When the tissue sample is mortared to powder, transfer it to an EP tube and add protein lysate.

[0050] (2) Lysis: Protease inhibitors and phosphatase inhibitors should be added to the ly...

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Abstract

The invention belongs to the field of biopharmaceuticals, and relates to a marker for diagnosing liver cancer, and a detection reagent and application of the marker, in particular to an application ofTRAF2 to diagnosis and treatment of the liver cancer. The invention provides the marker for diagnosing the liver cancer, the marker is the TRAF2, and the expression of the TRAF2 in liver cancer patients is raised. The invention provides a new biologic marker TRAF2, for diagnosing the liver cancer, besides, the TRAF2 can be used as a novel target point for treating the liver cancer, and a new direction is provided for screening medicines for diagnosing and treating the liver cancer and treating the liver cancer.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to markers for diagnosing liver cancer, detection reagents and applications thereof, in particular to the application of TRAF2 in the diagnosis and treatment of liver cancer. Background technique [0002] Primary liver cancer (PLC) is one of the few tumors whose incidence continues to rise. Worldwide, liver cancer ranks fourth among cancer-related deaths. China is a big country with liver cancer. According to the statistics of the World Health Organization, in 2018, there were 392,868 new cases of liver cancer (9.2%) and 368,960 deaths (12.9%) in China. Although some progress has been made in the diagnosis and treatment of liver cancer, due to the insidious onset of liver cancer, the early symptoms are often atypical, and the disease is often in the advanced stage when seeing a doctor. Therefore, the incidence of liver cancer is not only increased, but the overall survival rate of liver can...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/11A61K45/00A61P35/00
CPCC12Q1/6886A61K45/00A61P35/00C12Q2600/158C12Q2600/136
Inventor 郑敏梁雪赵永超刘艳宁姚吉平
Owner ZHEJIANG UNIV
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