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Method for regulating expression of vascular cell adhesion molecule-1 in vascular endothelial cell

A vascular endothelium, adhesion molecule technology, applied in the field of oncology, can solve problems such as little known

Active Publication Date: 2015-04-01
SHANGHAI INST OF ONCOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, to date, little is known in the art about how the expression or activity of vascular cell adhesion molecule-1 (VCAM-1) is regulated

Method used

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  • Method for regulating expression of vascular cell adhesion molecule-1 in vascular endothelial cell
  • Method for regulating expression of vascular cell adhesion molecule-1 in vascular endothelial cell
  • Method for regulating expression of vascular cell adhesion molecule-1 in vascular endothelial cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0126] Secreted ANGPTL4 protein up-regulates the expression of VCAM-1 in HUVEC cells

[0127] Conditioned medium (CM) supernatant was collected to treat liver cancer cells with low expression of ANGPTL4, and the effect of secreted ANGPTL4 protein on the migration of liver cancer cells across vascular endothelial cells was detected.

[0128] The CM used in this experiment was derived from the cell line COS7 stably overexpressing ANGPTL4, abbreviated as COS7-lenti-ANGPTL4, and the selection of COS7 cells can exclude the influence of other secreted factors of tumor cells.

[0129] Firstly, the expression level of ANGPTL4 in COS7-lenti-ANGPTL4 and its control group cells was verified. Compared with the control group, the expression level of ANGPTL4 in COS7-lenti-ANGPTL4 was significantly increased in both cell lysate and CM ( figure 1 a)

[0130] HUVEC cells were incubated with COS7-lenti-ANGPTL4 conditional culture supernatant for 24 hours, and the changes of VCAM-1 expression i...

Embodiment 2

[0134] Interfering with VCAM-1 or integrin β1 can partially inhibit transvascular endothelial cell migration induced by overexpression of ANGPTL4

[0135] The above studies indicated that secreted ANGPTL4 protein can upregulate the expression of VCAM-1 in HUVEC. In order to further determine the role of VCAM-1 in ANGPTL4 promoting transvascular endothelial cell migration, this example uses siRNA to interfere with the expression of VCAM-1 in HUVEC, and detects its effect on SMMC-7721-lenti-ANGPTL4 cells and SMMC-7721 Influence of -lenti-control cells on the ability to migrate across vascular endothelial cells.

[0136] The results showed that: SMMC-7721-lenti-ANGPTL4 had stronger transvascular endothelial cell migration ability than SMMC-7721-lenti-control, but the migration ability of SMMC-7721-lenti-ANGPTL4 was significantly reduced after interfering with VCAM-1 in HUVEC ( P0.05)( figure 2 a, b).

[0137] This example also verified the role of integrin β1 in ANGPTL4 promo...

Embodiment 3

[0143] Promote the expression of VCAM-1 with recombinant ANGPTL4 protein

[0144] Repeat Example 1, the difference is that the conditional medium (CM) supernatant is replaced with recombinant ANGPTL4 protein.

[0145] The results were the same as in Example 1, the recombinant ANGPTL4 protein (100ng / ml) could significantly up-regulate and promote the expression of VCAM-1 in vascular endothelial cells.

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Abstract

The invention relates to a method for regulating expression of vascular cell adhesion molecule-1 in a vascular endothelial cell. Specifically, the invention provides a method for regulating the expression of the vascular cell adhesion molecule-1 (VCAM-1) through an angiopoietin-like protein 4 (ANGPTL4 protein). The invention also provides application of the ANGPTL4 protein in preparing promoters promoting (VCAM-1) expression.

Description

technical field [0001] The present invention relates to the field of oncology. More specifically, the present invention relates to a method for regulating the expression of vascular cell adhesion molecule-1 in vascular endothelial cells through angiopoietin-like protein 4 (ANGPTL4 protein). 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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
Inventor 李锦军李红葛超顾健人
Owner SHANGHAI INST OF ONCOLOGY
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