Ectopic and eutopic stromal endometriotic cells have a damaged ceramide signaling pathway to apoptosis.
OBJECTIVE
To investigate whether sphingosine analogues, which activate the ceramide signaling pathway to apoptosis, can cause the death of ectopic (EEC) and eutopic stromal endometriotic cells (EEU), as well as healthy eutopic stromal endometrial cells (HEU).


METHODS
The EEC, EEU, and HEU isolated from fertile and infertile women with endometriosis were cultured for 48 hours in RPMI medium with 10% fetal calf serum (FCS) and with 2.5-10 microM sphingosine analogues.


METHODS
A clinic for the treatment of endometriosis and basic research laboratories.


METHODS
Nineteen women with follicular cyst and 16 women with endometriosis.


METHODS
The percentage of proliferating cells was determined by 93-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide (MTT) assay. Apoptosis and cell cycle were detected by fluorescence-activated cell sorter (FACS) Calibur flow cytometer.


RESULTS
The viability of EEC after exposure to 10 microM sphingosine analogues was 59.5% +/- 9.7% for D-sphingosine and 77.65 +/- 9.7% for DL-erythro-sphingosine, the viability of EEU was 69.2% +/- 14.2% and 42.0% +/- 15.5%, whereas the viability of comparative HEU was 9.0% +/- 4.8% and 18.8% +/- 8.3%, respectively. The differences were significant using the Mann-Whitney test. The apoptotic level of the cells treated with 10 microM sphingosine analogues for comparative HEU was 42.8% +/- 7.5% for D-sphingosine and 42.5% +/- 10.5% for DL-erythro-sphingosine, whereas for EEC this was 16.7% +/- 5.5% for D-sphingosine and 14.1% +/- 4.4% for DL-erythro-sphingosine and for EEU this was 14.3% +/- 4.7% and 22.9% +/- 8.9%, respectively.


CONCLUSIONS
Ectopic and eutopic stromal endometrial cells from women with endometriosis have a damaged ceramide-downstream pathway to apoptosis.
Chrobak
Agnieszka
A
Faculty of Biotechnology, University of Wroclaw, Wroclaw, Poland. chrobak@iitd.pan.wroc.pl
Sieradzka
Urszula
U
Sozański
Rafał
R
Chełmońska-Soyta
Anna
A
Gabryś
Marian
M
Jerzak
Małgorzata
M
D000328:Adult; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D001798:Blood Proteins / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D002518:Ceramides / Q000378:metabolism* / Q000494:pharmacology; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004715:Endometriosis / Q000378:metabolism* / Q000473:pathology; D004717:Endometrium / Q000166:cytology* / Q000473:pathology; D005260:Female; D016193:G1 Phase / Q000187:drug effects / Q000502:physiology; D016195:G2 Phase / Q000187:drug effects / Q000502:physiology; D006801:Humans; D016130:Immunophenotyping; D007247:Infertility, Female / Q000378:metabolism / Q000473:pathology; D008875:Middle Aged; D016192:Resting Phase, Cell Cycle / Q000187:drug effects / Q000502:physiology; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013110:Sphingosine / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism* / Q000473:pathology; D055815:Young Adult
D001798:Blood Proteins; D002518:Ceramides; D016895:Culture Media, Serum-Free; D013110:Sphingosine