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Application of serum DDK1 in preparation of diagnosis reagent for alpha fetal protein negative hepatocellular carcinoma

A technology of alpha-fetoprotein and diagnostic reagents, which is applied in the field of preparation of diagnostic reagents for alpha-fetoprotein-negative hepatocellular carcinoma, and can solve the problems of unclear diagnostic ability of patients with hepatocellular carcinoma

Inactive Publication Date: 2014-01-01
SHANGHAI INST OF ONCOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although there have been previous studies on DKK1, the clinical significance of serum DKK1, especially the diagnostic ability of AFP-negative (serum AFP≤20ng / ml) hepatocellular carcinoma patients is not clear in this field

Method used

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  • Application of serum DDK1 in preparation of diagnosis reagent for alpha fetal protein negative hepatocellular carcinoma
  • Application of serum DDK1 in preparation of diagnosis reagent for alpha fetal protein negative hepatocellular carcinoma
  • Application of serum DDK1 in preparation of diagnosis reagent for alpha fetal protein negative hepatocellular carcinoma

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Experimental program
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Embodiment approach

[0059] As an embodiment of the present invention, the reagent is an anti-DKK1 antibody; more specifically, it is a monoclonal antibody or a polyclonal antibody.

[0060] Antibodies of the present invention can be prepared by various techniques known to those skilled in the art. For example, purified antigen can be administered to animals such as rabbits, mice, rats, etc. to induce polyclonal antibody production. Various adjuvants can be used to enhance the immune response, including but not limited to Freund's adjuvant and the like.

[0061] Antibodies of the invention may also be monoclonal antibodies. Such monoclonal antibodies can be prepared using hybridoma technology.

[0062] The antibody can be used in immunochemical techniques to detect the DKK1 level in the specimen, so as to diagnose AFP-negative hepatocellular carcinoma or judge the risk (susceptibility) of the disease.

[0063] As another embodiment of the present invention, the reagent is a primer for specifica...

Embodiment 1

[0143] Example 1, Comparative analysis of the expression of DKK1 and AFP in non-cancerous liver tissue and hepatocellular carcinoma tissue and corresponding serum

[0144] The present inventors compared and analyzed DKK1 and AFP in 12 cases of non-cancerous liver tissues (4 cases were normal human liver tissues, 8 cases were liver cirrhosis tissues) and 16 cases of hepatocellular carcinoma liver tissues (8 cases of patients with serum AFP≤20ng / ml, and the expression in serum AFP>20ng / ml of the other 8 cases). The results of semi-quantitative reverse transcription polymerase chain reaction (semi-qPCR) and real-time quantitative polymerase chain reaction (qRT-PCR) experiments showed that compared with AFP, DKK1 was more active in 4 cases of healthy controls (HC) and 8 cases. In cases of liver cirrhosis (liver corrosion, LC), the expression was almost undetectable, but it was highly expressed in most liver tissues of hepatocellular carcinoma ( figure 1 A-B, 1D-E).

[0145] Enz...

Embodiment 2

[0147]Example 2. Serum DKK1 is a serum protein marker for the diagnosis of hepatocellular carcinoma

[0148] The inventor according to figure 2 To evaluate the diagnostic ability of serum DKK1 protein in AFP-negative HCC.

[0149] Firstly, the expression of serum DKK1 and AFP in each group of the test set was analyzed. Such as image 3 As shown in A, the serum DKK1 protein level of hepatocellular carcinoma (HCC) group was significantly higher than that of healthy controls (HC), chronic hepatitis B (chronic hepatitis B, CHB) control group and liver cirrhosis (liver cirrhosis, LC) control group (P image 3 B). See Table 4 for the concentrations of serum DKK1 and AFP in each group.

[0150] Therefore, serum DKK1 may be used as a serum protein marker for the diagnosis of HCC.

[0151] Table 4. Concentrations of serum DKK1 protein and serum AFP protein in each group in test set and validation set

[0152]

[0153]

[0154] Abbreviations: DKK1: dickkopf-1; AFP: alpha-fet...

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Abstract

The invention relates to an application of serum DDK1 in the preparation of a diagnosis reagent for alpha fetal protein negative hepatocellular carcinoma. Through analysis of a large amount of hepatocellular carcinoma samples, the inventor analyzes the expression situations of DKK1 and AFP from the samples, and firstly discovers that serum DKK1 has a high sensitivity and specificity on diagnosis of AFP negative hepatocellular carcinoma.

Description

technical field [0001] The invention belongs to the field of biomedicine; more specifically, the invention relates to the use of serum DKK1 in preparing a diagnostic reagent for alpha-fetoprotein-negative hepatocellular carcinoma. Background technique [0002] Primary liver tumors include hepatocellular carcinoma (HCC), intrahepatic cholangiocarcinoma, mixed hepatocellular carcinoma, cystadenoma of the bile duct, hepatoblastoma, hemangioma, carcinoid, lymphoma, epithelioid vascular endothelium Tumor, squamous cell carcinoma, teratoma, and leiomyosarcoma, fibrosarcoma, malignant fibrous histiocytoma, rhabdomyosarcoma and other liver sarcomas. At present, the etiology of hepatocellular carcinoma has not been fully clarified. Its occurrence is a process involving multi-factors, multi-stages, and multi-gene accumulation of mutations. The main factors are viral infection, aflatoxin intake, alcoholism, water pollution, and liver disease. Helicobacter and liver cirrhosis. Hepatit...

Claims

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Application Information

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IPC IPC(8): G01N33/574C12Q1/68
CPCG01N33/57438C12Q1/6886C12Q2600/112C12Q2600/158G01N33/6893
Inventor 覃文新沈秋瑾顾健人杨胜利
Owner SHANGHAI INST OF ONCOLOGY
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