Recombinant H2 relaxin inhibits apoptosis and induces cell proliferation in cultured leiomyoma cells without affecting those in cultured normal myometrial cells.
OBJECTIVE
To evaluate the effects of human relaxin on proliferation and apoptosis in cultured human uterine leiomyoma cells and normal myometrial cells.


METHODS
In vitro experiment.


METHODS
Research laboratory at Kobe University Graduate School of Medicine.


METHODS
Nine patients undergoing hysterectomy for uterine leiomyoma.


METHODS
Cultured leiomyoma cells and normal myometrial cells were treated with human recombinant (rH2) relaxin.


METHODS
Human relaxin receptor LGR7 expressions in cultured leiomyoma cells and myometrial cells were evaluated by immunocytochemical staining. Cell proliferation, proliferating cell nuclear antigen-positive rate, and TUNEL-positive rate were assessed by MTS assay, immunocytochemistry, and TUNEL assay, respectively. Caspase-3 expression was evaluated by Western blot analysis.


RESULTS
LGR7 expression was observed both in cultured human leiomyoma cells and myometrial cells. Compared with untreated control cultures, treatment with rH2 relaxin increased the number of viable cultured leiomyoma cells and the proliferating cell nuclear antigen-positive rate in those cells but not in myometrial cells. Moreover, treatment with rH2 relaxin decreased the TUNEL-positive rate in cultured leiomyoma cells but not in myometrial cells. Similarly, Western blot analysis revealed that treatment with rH2 relaxin decreased the expression of caspase-3 in cultured leiomyoma cells but not in myometrial cells.


CONCLUSIONS
These results suggest that rH2 relaxin selectively inhibits apoptosis by down-regulating caspse-3 expression and induces proliferation in cultured human leiomyoma cells without affecting apoptosis or proliferation in normal myometrial cells.
Department of Obstetrics and Gynecology, Kobe University Graduate School of Medicine, Kobe, Japan.
Kaho
K
Suzuki
Koji
K
Nakabayashi
Ai Y
AY
Yamada
Romana S Z
RS
Lodhi
Rhoichi
R
Hazama
Yasuhiko
Y
Ebina
Hideto
H
Yamada
D000328:Adult; D017209:Apoptosis; D015153:Blotting, Western; D053148:Caspase 3 / Q000378:metabolism; D049109:Cell Proliferation; D015536:Down-Regulation; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D007889:Leiomyoma / Q000378:metabolism* / Q000473:pathology; D008875:Middle Aged; D009215:Myometrium / Q000378:metabolism*; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D018000:Receptors, Peptide / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012065:Relaxin / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014594:Uterine Neoplasms / Q000378:metabolism* / Q000473:pathology
D018809:Proliferating Cell Nuclear Antigen; C095068:RLN2 protein, human; C417543:RXFP1 protein, human; D043562:Receptors, G-Protein-Coupled; D018000:Receptors, Peptide; D011994:Recombinant Proteins; D012065:Relaxin; C505407:CASP3 protein, human; D053148:Caspase 3