Relaxin has anti-apoptotic effects on human trophoblast-derived HTR-8/SV neo cells.
The study was conducted to evaluate the effects of human relaxin on apoptosis in the human trophoblast derived HTR-8/SV neo cell line, which is a possible model of human extravillous trophoblasts (EVTs). HTR-8/SV neo cells, cultured in phenol red free RPMI1640 medium, were treated with different doses of human recombinant (rH2) relaxin in serum-deprived conditions. RT-PCR was used for evaluating relaxin receptor: RXFP1 and RXFP2 expression in HTR-8/SV neo cells. The cell death was examined by TUNEL assay. Furthermore, we investigated caspase-3, cleaved PARP and Bcl-2 expressions by Western blot analysis to recognize the translational effects of anti-apoptotic and pro-apoptotic proteins. RXFP1 and RXFP2 mRNA expression was observed in HTR-8/SV neo cells. Compared with untreated control cultures, treatment with rH2 relaxin, decreased TUNEL-positive rate in HTR-8/SV neo cells was observed. Western blot analysis revealed that treatment with rH2 relaxin decreased the expression of caspase-3 and cleaved PARP, but in contrast increased Bcl-2 expression in those cells. These results suggest that rH2 relaxin has anti-apoptotic effects on HTR8/SV neo cells by decreasing pro-apoptotic caspase-3 and cleaved PARP expression and up-regulating anti-apoptotic Bcl-2 expression.
Department of Obstetrics and Gynecology, Kobe University Graduate School of Medicine , Kobe , Japan.
Romana S Z
RS
Lodhi
Koji
K
Nakabayashi
Kaho
K
Suzuki
Ai Y
AY
Yamada
Rhoichi
R
Hazama
Yasuhiko
Y
Ebina
Hideto
H
Yamada
D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000032:analysis; D002460:Cell Line; D016895:Culture Media, Serum-Free; D005260:Female; D015870:Gene Expression; D006801:Humans; D020287:In Situ Nick-End Labeling; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D043562:Receptors, G-Protein-Coupled / Q000235:genetics; D018000:Receptors, Peptide / Q000235:genetics; D011994:Recombinant Proteins / Q000008:administration & dosage; D012065:Relaxin / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014327:Trophoblasts / Q000737:chemistry / Q000166:cytology* / Q000187:drug effects*
D016895:Culture Media, Serum-Free; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; C417543:RXFP1 protein, human; C472867:RXFP2 protein, human; D043562:Receptors, G-Protein-Coupled; D018000:Receptors, Peptide; D011994:Recombinant Proteins; D012065:Relaxin; D053148:Caspase 3