20-Hydroxyeicosatetraenoic acid is formed in response to EGF and is a mitogen in rat proximal tubule

Fangming Lin, Amelia Rios, J R Falck, Yuri Belosludtsev, Michal Laniado Schwartzman

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Abstract

Hydroxyeicosatetraenoic acid (20-HETE) is a major cytochrome P-450-arachidonic acid metabolite in the rat kidney, and its synthesis along the nephron is specifically localized to the proximal tubule, where receptor density for epidermal growth factor (EGF) is the highest. EGF stimulated endogenous 20-HETE formation in a concentration and time-dependent manner, i.e., from 1.6 to 2.6 ± 0.3 and 3.0 ± 0.6 pmol 20-HETE·mg-1·min-1 at 10-8 and 10-7 M EGF, respectively. The effect of 20-HETE on proximal tubular cell proliferation was examined using primary cultures of rat proximal tubular cells and proximal tubular-derived cell lines, LLC-PK1 and opossum kidney OK. In both cell lines, 20-HETE increased thymidine incorporation into DNA with maximal effect at 10-9 M. Addition of 20-HETE to serum-deprived LLC-PK1 or OK cells for 48 h caused a concentration-dependent increase in cell number with maximal effect at 10-9 M. This effect was specific, as structurally similar eicosanoids such as 20-COOH-arachidonic acid, 19(R)-HETE, and 19(S)-HETE did not increase cell number. In 4-day primary cultures of proximal tubular cells, EGF (10-9 M) and 20-HETE (10-9 M) increased bromodeoxyuridine (BrdU) incorporation by 40 and 28%, respectively. Addition of both resulted in a twofold increase in BrdU incorporation. Although 20-HETE synthesis in cultured cells is greatly diminished with time, significant picomolar concentrations can be obtained in 4-day cultures. Addition of 17-octadecynoic acid (17-ODYA), an inhibitor of 20-HETE synthesis, significantly inhibited EGF-stimulated BrdU incorporation. The demonstrations that EGF stimulates proximal tubular 20-HETE production and that the latter is a potent mitogen to these cells suggests that 20-HETE may act as a mediator of the EGF effect on cellular growth in the proximal tubule.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Renal Physiology
Volume269
Issue number6
StatePublished - 1995

Fingerprint

Mitogens
Epidermal Growth Factor
Bromodeoxyuridine
Arachidonic Acid
Cell Count
20-hydroxy-5,8,11,14-eicosatetraenoic acid
Kidney
Hydroxyeicosatetraenoic Acids
Opossums
Cell Line
Eicosanoids
Nephrons
Epidermal Growth Factor Receptor
Thymidine
Cytochrome P-450 Enzyme System
Cultured Cells
Cell Proliferation
DNA
Growth
Serum

Keywords

  • Arachidonic acid
  • Cytochrome P-450
  • Epidermal growth factor
  • LLC-PK cells
  • Opossum kidney cells

ASJC Scopus subject areas

  • Physiology

Cite this

20-Hydroxyeicosatetraenoic acid is formed in response to EGF and is a mitogen in rat proximal tubule. / Lin, Fangming; Rios, Amelia; Falck, J R; Belosludtsev, Yuri; Schwartzman, Michal Laniado.

In: American Journal of Physiology - Renal Physiology, Vol. 269, No. 6, 1995.

Research output: Contribution to journalArticle

Lin, Fangming ; Rios, Amelia ; Falck, J R ; Belosludtsev, Yuri ; Schwartzman, Michal Laniado. / 20-Hydroxyeicosatetraenoic acid is formed in response to EGF and is a mitogen in rat proximal tubule. In: American Journal of Physiology - Renal Physiology. 1995 ; Vol. 269, No. 6.
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T1 - 20-Hydroxyeicosatetraenoic acid is formed in response to EGF and is a mitogen in rat proximal tubule

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AU - Rios, Amelia

AU - Falck, J R

AU - Belosludtsev, Yuri

AU - Schwartzman, Michal Laniado

PY - 1995

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N2 - Hydroxyeicosatetraenoic acid (20-HETE) is a major cytochrome P-450-arachidonic acid metabolite in the rat kidney, and its synthesis along the nephron is specifically localized to the proximal tubule, where receptor density for epidermal growth factor (EGF) is the highest. EGF stimulated endogenous 20-HETE formation in a concentration and time-dependent manner, i.e., from 1.6 to 2.6 ± 0.3 and 3.0 ± 0.6 pmol 20-HETE·mg-1·min-1 at 10-8 and 10-7 M EGF, respectively. The effect of 20-HETE on proximal tubular cell proliferation was examined using primary cultures of rat proximal tubular cells and proximal tubular-derived cell lines, LLC-PK1 and opossum kidney OK. In both cell lines, 20-HETE increased thymidine incorporation into DNA with maximal effect at 10-9 M. Addition of 20-HETE to serum-deprived LLC-PK1 or OK cells for 48 h caused a concentration-dependent increase in cell number with maximal effect at 10-9 M. This effect was specific, as structurally similar eicosanoids such as 20-COOH-arachidonic acid, 19(R)-HETE, and 19(S)-HETE did not increase cell number. In 4-day primary cultures of proximal tubular cells, EGF (10-9 M) and 20-HETE (10-9 M) increased bromodeoxyuridine (BrdU) incorporation by 40 and 28%, respectively. Addition of both resulted in a twofold increase in BrdU incorporation. Although 20-HETE synthesis in cultured cells is greatly diminished with time, significant picomolar concentrations can be obtained in 4-day cultures. Addition of 17-octadecynoic acid (17-ODYA), an inhibitor of 20-HETE synthesis, significantly inhibited EGF-stimulated BrdU incorporation. The demonstrations that EGF stimulates proximal tubular 20-HETE production and that the latter is a potent mitogen to these cells suggests that 20-HETE may act as a mediator of the EGF effect on cellular growth in the proximal tubule.

AB - Hydroxyeicosatetraenoic acid (20-HETE) is a major cytochrome P-450-arachidonic acid metabolite in the rat kidney, and its synthesis along the nephron is specifically localized to the proximal tubule, where receptor density for epidermal growth factor (EGF) is the highest. EGF stimulated endogenous 20-HETE formation in a concentration and time-dependent manner, i.e., from 1.6 to 2.6 ± 0.3 and 3.0 ± 0.6 pmol 20-HETE·mg-1·min-1 at 10-8 and 10-7 M EGF, respectively. The effect of 20-HETE on proximal tubular cell proliferation was examined using primary cultures of rat proximal tubular cells and proximal tubular-derived cell lines, LLC-PK1 and opossum kidney OK. In both cell lines, 20-HETE increased thymidine incorporation into DNA with maximal effect at 10-9 M. Addition of 20-HETE to serum-deprived LLC-PK1 or OK cells for 48 h caused a concentration-dependent increase in cell number with maximal effect at 10-9 M. This effect was specific, as structurally similar eicosanoids such as 20-COOH-arachidonic acid, 19(R)-HETE, and 19(S)-HETE did not increase cell number. In 4-day primary cultures of proximal tubular cells, EGF (10-9 M) and 20-HETE (10-9 M) increased bromodeoxyuridine (BrdU) incorporation by 40 and 28%, respectively. Addition of both resulted in a twofold increase in BrdU incorporation. Although 20-HETE synthesis in cultured cells is greatly diminished with time, significant picomolar concentrations can be obtained in 4-day cultures. Addition of 17-octadecynoic acid (17-ODYA), an inhibitor of 20-HETE synthesis, significantly inhibited EGF-stimulated BrdU incorporation. The demonstrations that EGF stimulates proximal tubular 20-HETE production and that the latter is a potent mitogen to these cells suggests that 20-HETE may act as a mediator of the EGF effect on cellular growth in the proximal tubule.

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