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Research method of Crosstalk mechanism of VEGF-CXCL8-Akt and application thereof

A vegf-cxcl8-akt, mechanism technology, applied in biochemical equipment and methods, compound screening, microbial assay/test, etc., can solve problems such as poor therapeutic effect

Pending Publication Date: 2021-02-19
NINGXIA MEDICAL UNIV +1
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  • Application Information

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Problems solved by technology

[0004] Glioma is the most common primary tumor of the central nervous system, and the current treatment effect is poor

Method used

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  • Research method of Crosstalk mechanism of VEGF-CXCL8-Akt and application thereof
  • Research method of Crosstalk mechanism of VEGF-CXCL8-Akt and application thereof
  • Research method of Crosstalk mechanism of VEGF-CXCL8-Akt and application thereof

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Experimental program
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Embodiment Construction

[0033] The technical solutions of the present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0034] 1 Materials and methods

[0035] 1.1 Main reagent materials

[0036] Human glioma cell lines U87MG and U251 were purchased from the Shanghai Cell Resource Center of the Chinese Academy of Sciences. Both are glioblastoma monoclonal lines, and the WHO grading standard is IV. Cultured in DMEM medium containing 10% fetal bovine serum (DMEM medium, trypsin were purchased from Gibco, USA, and fetal bovine serum was purchased from HyClone, USA). Human brain microvascular endothelial cells (human brain microvascular endothelial cells, HBMEC) were purchased from Sciencell, USA, and cultured in ECM medium containing 10% fetal bovine serum (ECM medium and fetal bovine serum were purchased from Sciencell, USA). VEGF 165 , rhCXCL8 was purchased from Pepro Tech Company of the United States, and LY294002 was purc...

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Abstract

The invention discloses a research method of a Crosstalk mechanism of VEGF-CXCL8-Akt and application thereof. The research method comprises the following steps of firstly, detecting the influence of U87MG, U251 and secretion factors VEGF thereof on secretion of CXCL8 by HBMEC by using ELISA, secondly, respectively adding a specific agonist CXCL8 and an inhibitor LY294002 of a CXCL8-Akt signal pathinto a co-culture system, and detecting the influence of co-culture and activation / inhibition on expression of Akt proteins of U87MG and U251 cells and phosphorylation activation state P-Akt thereofby using Western blotting, and then using CCK-8 and Transwell invasion experiments for respectively detecting the influence of co-culture and activation / inhibition on proliferation and invasion of U87MG and U251 cells.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to a research method and application of the Crosstalk mechanism of VEGF-CXCL8-Akt, in particular, to a method for detecting VEGF between glioma cells and vascular endothelial cells from the perspective of signal network. -Research methods and applications for the study of the Crosstalk mechanism of CXCL8-Akt. Background technique [0002] Glioma is a solid tumor with strong dependence on blood vessels. When the tumor grows to a diameter of more than 1-2 mm, it needs peripheral blood vessels to provide additional support. Therefore, the interaction mechanism between tumors and surrounding blood vessels is very important. Studies have confirmed that angiogenesis factors are an important bridge in the signaling crosstalk mechanism between glioma cells and endothelial cells, and vascular endothelial growth factor (VEGF) is one of the most important factors. The increase of VEGF expre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/573G01N21/31C12Q1/02
CPCG01N33/6863G01N33/573G01N21/31G01N33/5032G01N33/5064G01N33/5044G01N2500/10
Inventor 顾金海杨景煜孙涛牛建国王峰文玉军王鹏强媛媛和祯泉杨勇
Owner NINGXIA MEDICAL UNIV
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