Selective progesterone receptor modulator asoprisnil induces endoplasmic reticulum stress in cultured human uterine leiomyoma cells.
A recent clinical trial (Chwalisz K, Larsen L, Mattia-Goldberg C, Edmonds A, Elger W, Winkel CA. Fertil Steril 87: 1399-1412, 2007) has demonstrated that the selective progesterone receptor modulator asoprisnil efficiently causes the shrinkage of uterine leiomyoma. The present study was conducted to examine whether asoprisnil elicits endoplasmic reticulum (ER) stress-induced apoptosis in cultured human uterine leiomyoma cells. After subculture in phenol red-free DMEM supplemented with 10% FBS for 120 h, cultured cells were stepped down to serum-free conditions with or without graded concentrations of asoprisnil. ER stress-associated and apoptosis-related proteins were assessed by reverse transcription-PCR analysis or Western blot analysis. RNA interference of growth-arrest- and DNA-damage-inducible gene 153 (GADD153) was performed using small interfering RNA. Terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate nick end labeling (TUNEL)-positive rates were assessed by TUNEL assay. Compared with untreated control cultures, treatment with 10(-7) M asoprisnil significantly (P < 0.05) increased the protein contents of ubiquitin at 2 h and phospho-double-stranded RNA-activated protein kinase-like ER kinase, phospho-eukaryotic initiation factor 2alpha, activating transcription factor 4, and glucose-regulated protein 78 kDa at 4 h, followed by the significant (P < 0.05) increase in GADD153 protein content at 6 h and cleaved poly(adenosine 5'-diphosphate ribose)polymerase (PARP) at 8 h. RNA interference of GADD153 suppressed protein contents of asoprisnil-induced cleaved PARP, Bax, Bak, GADD34, and tribbles-related protein 3 (TRB3) and TUNEL-positive rate but attenuated asoprisnil-induced reduction in Bcl-2 protein content in cultured leiomyoma cells. These results suggest that asoprisnil elicits ER stress-induced apoptosis in cultured leiomyoma cells and that GADD153 plays a role in asoprisnil-induced apoptosis by modulating the Bcl-2 family of proteins, GADD34, and TRB3.
Xu
Qin
Q
Dept. of Obstetrics and Gynecology, Kobe Univ. Graduate School of Medicine, 7-5-1 Kusunoki-Cho, Chuo-Ku, Kobe, Hyogo Prefecture, 650-0017, Japan.
Ohara
Noriyuki
N
Liu
Jin
J
Nakabayashi
Koji
K
DeManno
Deborah
D
Chwalisz
Kristof
K
Yoshida
Shigeki
S
Maruo
Takeshi
T
D000328:Adult; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D004721:Endoplasmic Reticulum / Q000187:drug effects* / Q000378:metabolism; D004963:Estrenes / Q000494:pharmacology*; D005260:Female; D006801:Humans; D007889:Leiomyoma / Q000378:metabolism / Q000473:pathology*; D008875:Middle Aged; D010091:Oximes / Q000494:pharmacology*; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D045167:Progesterone Congeners / Q000494:pharmacology; D017510:Protein Folding; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D011980:Receptors, Progesterone / Q000819:agonists*; D013379:Substrate Specificity; D051742:Transcription Factor CHOP / Q000235:genetics / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014415:Tunicamycin / Q000494:pharmacology; D025801:Ubiquitin / Q000378:metabolism; D014594:Uterine Neoplasms / Q000378:metabolism / Q000473:pathology*
C497249:DDIT3 protein, human; D004963:Estrenes; D010091:Oximes; D045167:Progesterone Congeners; D011980:Receptors, Progesterone; D025801:Ubiquitin; D014415:Tunicamycin; D051742:Transcription Factor CHOP; C488516:asoprisnil; D011065:Poly(ADP-ribose) Polymerases