Interleukin-8 gene and protein expression are up-regulated by interleukin-1beta in normal human ovarian cells and a granulosa tumor cell line.
OBJECTIVE
To evaluate the expression, regulation, and role of interleukin (IL)-8 in human ovary.


METHODS
Prospective study.


METHODS
University hospital.


METHODS
Sixteen premenopausal women.


METHODS
Follicular fluid and granulosa lutein cells (GLCs) were collected during IVF cycles. Ovarian stromal and theca cells were obtained from women underwent surgery. KGN cells, the human granulosa cell tumor cell line, were also used.


METHODS
The levels of IL-8 and IL-1beta in follicular fluid and IL-8 protein production were determined using ELISA. Interleukin-8 and IL-8 receptor gene expression in ovarian cells and the effect of IL-8 on the proliferation of stromal cells were determined. The expression of pIkappaB was evaluated by Western blot, and the effect of NF-kappaB inhibitor APDC was examined by Northern blot analysis and ELISA in KGN cells. The levels of IL-8 and IL-1beta in follicular fluid; each concentration and the volume showed a positive correlation. Reverse transcription polymerase chain reaction showed the presence of IL-8 mRNA in all ovarian cells. In contrast, IL-8 receptor mRNA was only detected in stromal cells. The expression of IL-8 in GLCs and KGN cells was increased by addition of IL-1beta and TNFalpha. Interleukin-8 increased the proliferation of ovarian stromal cells. The expression of pIkappaB in KGN cells was induced by IL-1beta, and the effects were reduced by APDC.


CONCLUSIONS
Interleukin 8 induced by IL-1beta via activation of NF-kappaB in granulosa cells may have a role in the periovulatory period of follicular maturation.
Fujii
Akiko
A
Department of Obstetrics and Gynecology, Tottori University School of Medicine, Yonago, Japan.
Harada
Tasuku
T
Yamauchi
Nobuhiro
N
Iwabe
Tomio
T
Nishi
Yoshihiro
Y
Yanase
Toshihiko
T
Nawata
Hajime
H
Terakawa
Naoki
N
D015152:Blotting, Northern; D015153:Blotting, Western; D002455:Cell Division; D002478:Cells, Cultured; D005260:Female; D015571:Follicular Fluid / Q000737:chemistry; D005786:Gene Expression Regulation; D006106:Granulosa Cell Tumor / Q000378:metabolism*; D006107:Granulosa Cells / Q000378:metabolism; D006801:Humans; D020672:I-kappa B Proteins / Q000032:analysis; D007375:Interleukin-1 / Q000032:analysis / Q000494:pharmacology*; D016209:Interleukin-8 / Q000032:analysis* / Q000235:genetics* / Q000494:pharmacology; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000502:physiology; D010051:Ovarian Neoplasms / Q000378:metabolism*; D010053:Ovary / Q000378:metabolism*; D010766:Phosphorylation; D023062:Receptors, Interleukin-8A / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017154:Stromal Cells / Q000166:cytology; D014407:Tumor Cells, Cultured
D020672:I-kappa B Proteins; D007375:Interleukin-1; D016209:Interleukin-8; D016328:NF-kappa B; D023062:Receptors, Interleukin-8A