TNFalpha gene silencing reduced lipopolysaccharide-promoted proliferation of endometriotic stromal cells.
OBJECTIVE
We previously reported that lipopolysaccharide (LPS)-promoted endometriotic stromal cell (ESC) proliferation by inducing TNFalpha production. The aim of this study was to investigate the efficacy of TNFalpha gene silencing on LPS-treated ESCs.


METHODS
Endometriotic stromal cells (ESCs) and endometrial stromal cells (ESCs) (EMSCs) were obtained from ovarian chocolate cysts and uterine myoma, respectively. Using PCR array, LPS-induced gene expression profiling after transfection of TNFalpha siRNA into ESCs was performed. Down-regulated genes by TNFalpha silencing were examined using real-time RT-PCR. Effect of TNFalpha silencing was examined using ELISA and BrdU incorporation, respectively.


RESULTS
In PCR array, TNFalpha silencing in ESCs repressed LPS-induced expression of cIAP2 and IL-8, NFkappaB pathway responsive genes. After adding LPS, the levels of cIAP2 and IL-8 expression in ESCs were higher compared with those in EMSCs. TNFalpha silencing attenuated the LPS-induced ESC proliferation.


CONCLUSIONS
Tumor necrosis factor alpha may be involved in cell proliferation of endometriotic tissues.
Miyamoto
Ayako
A
Department of Obstetrics and Gynecology, Tottori University School of Medicine, Nishimachi, Yonago, Japan.
Taniguchi
Fuminori
F
Tagashira
Yukiko
Y
Watanabe
Ayako
A
Harada
Tasuku
T
Terakawa
Naoki
N
D017209:Apoptosis / Q000276:immunology; D000075747:Baculoviral IAP Repeat-Containing 3 Protein; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D004715:Endometriosis / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D004717:Endometrium / Q000473:pathology; D005260:Female; D020869:Gene Expression Profiling; D020868:Gene Silencing; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D016209:Interleukin-8 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D007889:Leiomyoma / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D008070:Lipopolysaccharides / Q000494:pharmacology; D016328:NF-kappa B / Q000276:immunology; D010048:Ovarian Cysts / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D010049:Ovarian Diseases / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D034741:RNA, Small Interfering; D015398:Signal Transduction / Q000276:immunology; D017154:Stromal Cells / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics* / Q000276:immunology; D044767:Ubiquitin-Protein Ligases; D014594:Uterine Neoplasms / Q000235:genetics / Q000276:immunology* / Q000473:pathology
D051034:Inhibitor of Apoptosis Proteins; D016209:Interleukin-8; D008070:Lipopolysaccharides; D016328:NF-kappa B; D034741:RNA, Small Interfering; D014409:Tumor Necrosis Factor-alpha; C494832:BIRC3 protein, human; D000075747:Baculoviral IAP Repeat-Containing 3 Protein; D044767:Ubiquitin-Protein Ligases