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ANGPTL4 as a marker of liver cancer metastasis by serological detection and application thereof

A technology for liver cancer metastasis and detection of markers, applied in the field of oncology and diagnosis, can solve the problems of lack of serum markers, lack of serum markers of liver cancer metastasis, etc.

Active Publication Date: 2015-06-10
SHANGHAI INST OF ONCOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing detection of cancer (such as HCC) still lacks satisfactory serum markers, and there is even a lack of serum markers that can be used to detect or judge liver cancer metastasis

Method used

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  • ANGPTL4 as a marker of liver cancer metastasis by serological detection and application thereof
  • ANGPTL4 as a marker of liver cancer metastasis by serological detection and application thereof
  • ANGPTL4 as a marker of liver cancer metastasis by serological detection and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0155] 1. ANGPTL4 is highly expressed in HCC cell lines with high metastatic potential

[0156] Quantitative PCR and Western blot were used to detect the expression levels of ANGPTL4 mRNA and protein in a variety of commonly used liver cancer cell lines.

[0157] The result is as figure 1shown in a and 1c. ANGPTL4 was relatively highly expressed in human high metastatic potential liver cell lines MHCC-LM3, MHCC-97, MHCC-97L and MHCC-97H, while in other non-high metastatic liver cancer cell lines HepG2, Hep3B, Huh-7, Low expression of SMMC-7721, FOCUS and PLC / PRF / 5.

[0158] Since ANGPTL4 is a secreted protein, ELISA was also used to detect whether the secreted ANGPTL4 protein level was consistent with the expression level of the cells themselves.

[0159] The results showed that the secretion level of secreted sANGPTL4 in the cell culture supernatant was consistent with the trend of Western blot detection results ( figure 1 b).

[0160] The above results indicated that th...

Embodiment 2

[0162] Exogenous expression of ANGPTL4 promotes trans-endothelial migration of liver cancer cells

[0163] In this example, according to the expression of ANGPTL4 in various liver cancer cell lines, the liver cancer cells Huh7 and SMMC-7721 with low expression of ANGPTL4 were selected, and the cell lines with stable overexpression of ANGPTL4 were constructed using lentiviral vectors, which were named Huh7-lenti-ANGPTL4 and SMMC-7721-lenti-ANGPTL4, the corresponding controls were named Huh7-lenti-control and SMMC-7721-lenti-control.

[0164] The expression levels of ANGPTL4 in ANGPTL4 stable overexpression cell lines were detected by quantitative PCR and Western blot respectively ( figure 2 a, b), Successful overexpression of ANGPTL4 was confirmed.

[0165] In order to verify the expression of secreted ANGPTL4 protein, the level of sANGPTL4 protein in the cell culture supernatant of ANGPTL4 stable overexpression cell line was detected by ELISA and Western blot ( figure 2 a,...

Embodiment 3

[0168] Exogenous expression of ANGPTL4 promotes intrahepatic and lung metastasis of SMMC-7721 cells in nude mice

[0169] In vitro experiments have confirmed that ANGPTL4 can promote the transvascular endothelial cell migration of liver cancer cells. This example further studies the effect of ANGPTL4 on the metastasis of liver cancer cells in vivo.

[0170] The method is as follows: Nude mice were orthotopically inoculated with SMMC-7721-lenti-ANGPTL4 cells and corresponding control group SMMC-7721-lenti-control cells, and the experimental animals were sacrificed painlessly 6 weeks after the operation, and serum, liver and lung tissues were collected. Routine histopathological examination was performed on the liver and lung tissues, and the number of nodules with intrahepatic metastasis and lung metastasis in the nude mice of the experimental group and the control group were counted.

[0171] The results showed that although all the nude mice in the experimental group and the ...

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Abstract

The invention relates to ANGPTL4 as a marker of liver cancer metastasis by serological detection and an application thereof. Specifically, the invention provides an application of angiopoietin-like protein 4(ANGPTL4 protein) in (a) the preparation of a diagnostic reagent or kit for the detection of liver cancer metastasis and (b) the preparation of a diagnostic reagent or kit for serological detection of liver cancer. The invention also provides a related detection kit. The invention also provides a method for inhibiting hepatoma cell metastasis by the use of an antagonist of the ANGPTL 4 protein.

Description

technical field [0001] The present invention relates to the fields of oncology and diagnostics. More specifically, the present invention relates to a serum marker that can be used to detect liver cancer metastasis and its application. Background technique [0002] ANGPTL4 gene (initially the gene was named pp1158 by the inventor's laboratory) [Zhu Hongxin et al., Chinese Journal of Oncology (2002) 24 (2): 123-125] is to use this functional screening technology platform to clone from the human placenta cDNA library. The full-length cDNA of this gene is 1943bp, the open reading frame contains 1218bp, encodes 406 amino acids, and the estimated molecular weight is 45.2kDa. There is a hydrophobic signal peptide and a coiled-coil domain at the N-terminus, and a Fibrinogen-like domain at the C-terminus. And in 2000, the cDNA sequence of the gene (the original gene name was pp1158) was registered on GenBank (accession number is AF202636). PGAR [Mol.Cell.Biol., Jul 2000; 20:5343-5...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574G01N33/532
Inventor 李锦军李红姚明葛超闫明霞赵方瑜田华郝向芳顾健人
Owner SHANGHAI INST OF ONCOLOGY
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