Use of Smad3 protein in peripheral blood exosome as molecular marker and liver cancer detection kit

A detection kit and molecular marker technology, applied in the field of biotechnology detection, can solve problems such as imperfection, death, and difficulty in treatment

Active Publication Date: 2018-07-24
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In addition, local treatment methods such as surgery are currently used for liver cancer, and postoperative recurrence is an important cause of difficult follow-up treatment and

Method used

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  • Use of Smad3 protein in peripheral blood exosome as molecular marker and liver cancer detection kit
  • Use of Smad3 protein in peripheral blood exosome as molecular marker and liver cancer detection kit
  • Use of Smad3 protein in peripheral blood exosome as molecular marker and liver cancer detection kit

Examples

Experimental program
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Effect test

Embodiment 1

[0028] Extraction and identification of exosomes from human peripheral blood. Methods as below:

[0029] Human peripheral blood was collected using anticoagulant blood collection tubes, and after centrifugation (3000g×5 minutes) to remove blood cells, the supernatant was frozen in a -80°C refrigerator. When testing, remove the frozen supernatant and thaw at 4°C. Centrifuge (10000g x 60 minutes) to remove large cell debris and other substances. Take 10 μl of supernatant, add 3 μl of Exoquick reagent (purchased from System Biosciences), and let stand at 4° C. for 30 minutes. Discard the supernatant after centrifugation (3000g × 30 minutes), and discard the supernatant after centrifugation again (3000g × 5 minutes), and the obtained precipitate is exosomes.

Embodiment 2

[0031] The enzyme-linked immunosorbent assay (ELISA) was used to detect the content of exosome Smad3 in peripheral blood. The specific steps were as follows: exosomes were extracted from the peripheral blood of patients, and the protein was obtained after repeated freezing and thawing three times. The Smad3 protein content was detected according to the method steps of the Smad3 protein ELISA kit (purchased from Mlbio). The AFP data comes from the detection data generated during the clinical diagnosis and treatment of patients. The Laboratory Department of the Second Hospital of Zhejiang Medical University determined the AFP content according to the method steps of the alpha-fetoprotein assay kit (purchased from Architect).

Embodiment 3

[0033] The method described in Example 2 was used to detect the Smad3 protein content in peripheral blood exosomes of patients with liver cancer and obtain corresponding AFP data, and the peripheral blood of healthy people was used as a control. The AFP data of corresponding patients and healthy people come from the detection data generated during their clinical diagnosis and treatment.

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Abstract

The invention discloses use of a Smad3 protein in a peripheral blood exosome as a molecular marker for preparing a liver cancer detection kit and the liver cancer detection kit. A research of the invention shows that the Smad3 protein content of an exosome extracted from human peripheral blood is obviously increased in a liver cancer patient; the Smad3 protein used as a tumor marker for diagnosinga liver cancer obtains an area under the ROC curve of 0.888; in combination with an alpha fetal protein for diagnosing the liver cancer, the area under the ROC curve can be up to 0.975; therefore, the Smad3 in the peripheral blood exosome can be independently used as the tumor marker for diagnosing the liver cancer or can be jointed with the alpha fetal protein to be used as the tumor marker fordiagnosing the liver cancer, and also can be used for judging a postoperative recurrence risk of a liver cancer patient.

Description

technical field [0001] The invention relates to the technical field of biotechnology detection, in particular to the application of Smad3 protein in peripheral blood exosomes as a molecular marker and a liver cancer detection kit. Background technique [0002] The incidence of HCC in my country accounts for more than half of the world. Liver cancer has a high degree of malignancy and poor prognosis, with a 5-year survival rate of only 16.8%. [0003] One of the main reasons for the poor prognosis of liver cancer is that the onset of liver cancer is hidden. Most patients are found in the late stage, and there is no effective drug treatment for the late stage. Therefore, developing new tumor markers to help early diagnosis of HCC is crucial to improve the prognosis of HCC patients. Alpha-fetoprotein (Alpha-fetal protein, AFP) is a clinically widely used tumor marker for liver cancer. Although the sensitivity and specificity of AFP in diagnosing liver cancer are high, there ...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/68
CPCG01N33/57438G01N33/57488G01N33/6893
Inventor 梁廷波白雪莉章琦傅琦涵
Owner ZHEJIANG UNIV
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