Obstet Gynecol Sci.  2018 Jan;61(1):71-78. 10.5468/ogs.2018.61.1.71.

Effects of estradiol on HIF-1α expression and trophoblast differentiation in first trimester villous explant cultures

• Cho GJ
• Lee LH
• Lee B
• Lee J
• Ahn KH
• Hong SC
• Kim HJ
• Oh MJ

• KMID: 2420154
• DOI: http://doi.org/10.5468/ogs.2018.61.1.71

Abstract

OBJECTIVE
The purpose of this study was to investigate the effects of estradiol on the expression of hypoxia-inducible factor (HIF)-1α and the differentiation of trophoblasts in human first trimester villous explant cultures.
METHODS
Villous explant cultures were established from first trimester human placentas (6-8 weeks of gestation, n=3). Normal villous tissues were explanted on Matrigel and incubated under 3% O2 tension for 5 days. To evaluate the effects of estradiol on the villous explant cultures, 1 ng/mL of estradiol was added to the culture medium. The morphological integrities and viabilities of the villous explants were monitored. Immunohistochemistry for α5 and α1 integrin was performed to assess differentiation of extravillous trophoblasts (EVTs). Expression of HIF-1α in villous explant cultures was evaluated by western blotting and densitometry.
RESULTS
EVTs emerging from first trimester villous explant cultures formed outgrowths of cells from the distal ends and invaded the surrounding Matrigel. Exposure of villous explants to estradiol resulted in the decreased outgrowth of cells from the distal end and decreased expression of α5 integrin. However, estradiol treatment increased the invasion of villous explants into the surrounding Matrigel, concomitant with the increased expression of α1 integrin, indicating differentiation of EVTs into more invasive EVTs. On western blots, the expression of HIF-1α decreased significantly after treatment with estradiol under 3% O2 tension.
CONCLUSION
Our findings suggest that estradiol may downregulate expression of HIF-1α in placenta, which in turn promote trophoblast differentiation into invasive phenotype.

Keyword

Estradiol; Hypoxia-inducible factor-1α; Trophoblasts

MeSH Terms

Blotting, Western
Densitometry
Estradiol*
Female
Humans
Immunohistochemistry
Phenotype
Placenta
Pregnancy
Pregnancy Trimester, First*
Trophoblasts*
Estradiol

Figure

• 

  Fig. 1 Effects of estradiol on extravillous trophoblast outgrowth and invasion in first trimester villous explant cultures. Dissected villous tissues were cultivated on Matrigel under 3% O2 tension (A, C) or under 3% O2 tension with 1 ng/mL of estradiol (B, D). A series of images were taken under an inverted microscope at ×10 magnification at day 0 (A, B) and day 5 (C, D).

• 

  Fig. 2 Immunolocalizations of α5 and α1 integrins in villous explants. Sections of explants cultured under 3% O2 tension (A, C) or under 3% O2 tension with 1 ng/mL of estradiol (B, D) were stained for α5 (A, B) or α1 integrin (C, D). Images were taken at ×40 magnification.

• 

  Fig. 3 Western blot analysis of hypoxia-inducible factor (HIF)-1α in villous explant cultures. (A) The expression of HIF-1α from gestational weeks 6 to 8 villous explants were cultured under 3% O2 tension with 1 ng/mL of estradiol for 5 days and were compared with that of control explant cultures. (B) Densitometric analysis. Data are expressed as a percentage of control values (explants cultured under 3% O2 tension) of 3 separate experiments in triplicate. GAPDH, glyceraldehyde 3-phosphate dehydrogenase. a)P<0.05 vs. control.

• 

  Fig. 4 Western blot analysis of hypoxia-inducible factor (HIF)-1α in villous explant cultures. (A) Expression of HIF-1α from villous explants cultured under 3% O2 tension with 0.1–10 ng/mL of estradiol for 5 days was compared with that of control explant cultures. (B) Densitometric analysis. Data are expressed as a percentage of control values (explants cultured under 3% O2 tension) of 3 separate experiments performed in triplicate. GAPDH, glyceraldehyde 3-phosphate dehydrogenase. a)P<0.05 vs. control.

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