The influence of hypoxia on the expression of VEGF and its receptors in trophoblast cells
The influence of hypoxia on the expression of VEGF and its receptors in trophoblast cells
This study examines the influence of hypoxia on the expression of VEGF and its receptors by the trophoblast derived cell lines JEG, JAR and BeWo. Further experiments went on to examine the above with cultured natural human trophoblast cells.
Cells were grown under normoxic and hypoxic conditions for varying time periods. The average oxygen tension in the culture media of the hypoxia cultures (6-7 kPa) was significantly less than in the normoxic cultures (19-21 kPa). RNA was extracted and messages for VEGF121, VEGF165 and VEGF189 were found in all cells lines by reverse transcription and the polymerase chain reaction (RT-PCR). All three spliced variants were upregulated by hypoxia; findings were confirmed by competitive PCR for VEGF and expression of a house keeping gene. In addition it was possible to show that the message for the VEGF receptor flt-1 was upregulated by hypoxia.
Hypoxia resulted in an increase in VEGF protein in culture media from the cell lines (p<0.05) as determined by radioimmunoassay. However it was not possible to detect radioligand binding of 1251-VEGF to these cells to confirm the presence of significant amounts of active receptor to elicit an autocrine response. Therefore, normal trophoblast cells were isolated to study the presence of VEGF receptors on their surface to assess any differences from the cell lines. Interestingly radioligand binding studies were successful with natural trophoblast cells and increased binding of 125I-VEGF was shown after the cells had been cultured under hypoxic conditions. This VEGF receptor did not have the reported binding characteristics of flt-1 since it had a lower affinity for VEGF and was possibly a previously detected but uncharacterised VEGF receptor. Thus, VEGF and its receptors may in part be regulated through oxygen supply, and be of paramount importance for successful embryo implantation and subsequent placental and fetal growth.
University of Southampton
1998
Taylor, Cheryl Mary
(1998)
The influence of hypoxia on the expression of VEGF and its receptors in trophoblast cells.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
This study examines the influence of hypoxia on the expression of VEGF and its receptors by the trophoblast derived cell lines JEG, JAR and BeWo. Further experiments went on to examine the above with cultured natural human trophoblast cells.
Cells were grown under normoxic and hypoxic conditions for varying time periods. The average oxygen tension in the culture media of the hypoxia cultures (6-7 kPa) was significantly less than in the normoxic cultures (19-21 kPa). RNA was extracted and messages for VEGF121, VEGF165 and VEGF189 were found in all cells lines by reverse transcription and the polymerase chain reaction (RT-PCR). All three spliced variants were upregulated by hypoxia; findings were confirmed by competitive PCR for VEGF and expression of a house keeping gene. In addition it was possible to show that the message for the VEGF receptor flt-1 was upregulated by hypoxia.
Hypoxia resulted in an increase in VEGF protein in culture media from the cell lines (p<0.05) as determined by radioimmunoassay. However it was not possible to detect radioligand binding of 1251-VEGF to these cells to confirm the presence of significant amounts of active receptor to elicit an autocrine response. Therefore, normal trophoblast cells were isolated to study the presence of VEGF receptors on their surface to assess any differences from the cell lines. Interestingly radioligand binding studies were successful with natural trophoblast cells and increased binding of 125I-VEGF was shown after the cells had been cultured under hypoxic conditions. This VEGF receptor did not have the reported binding characteristics of flt-1 since it had a lower affinity for VEGF and was possibly a previously detected but uncharacterised VEGF receptor. Thus, VEGF and its receptors may in part be regulated through oxygen supply, and be of paramount importance for successful embryo implantation and subsequent placental and fetal growth.
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Published date: 1998
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Local EPrints ID: 463183
URI: http://eprints.soton.ac.uk/id/eprint/463183
PURE UUID: 76069e1c-404f-4daa-bfb8-8d2422c46718
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Date deposited: 04 Jul 2022 20:47
Last modified: 04 Jul 2022 20:47
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Author:
Cheryl Mary Taylor
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