Selective progesterone receptor modulator asoprisnil induces endoplasmic reticulum stress in cultured human uterine leiomyoma cells

• Published in: American journal of physiology: endocrinology and metabolism 2007-10, Vol.293 (4), p.E1002-E1011
• Main Authors: Xu, Qin, Ohara, Noriyuki, Liu, Jin, Nakabayashi, Koji, DeManno, Deborah, Chwalisz, Kristof, Yoshida, Shigeki, Maruo, Takeshi
• Format: Article
• Language: English
• Subjects: Adult; Apoptosis - drug effects; Apoptosis - genetics; Dose-Response Relationship, Drug; Endocrine system; Endocrinology; Endoplasmic Reticulum - drug effects; Endoplasmic Reticulum - metabolism; Estrenes - pharmacology; Female; Gynecology; Humans; Leiomyoma - metabolism; Leiomyoma - pathology; Middle Aged; Oximes - pharmacology; Poly(ADP-ribose) Polymerases - metabolism; Progesterone; Progesterone Congeners - pharmacology; Protein Folding; Protein Processing, Post-Translational - drug effects; Receptors, Progesterone - agonists; Reproductive system; Substrate Specificity; Transcription Factor CHOP - genetics; Transcription Factor CHOP - metabolism; Tumor Cells, Cultured; Tumors; Tunicamycin - pharmacology; Ubiquitin - metabolism; Uterine Neoplasms - metabolism; Uterine Neoplasms - pathology
• ISSN: 0193-1849; 1522-1555
• DOI: 10.1152/ajpendo.00210.2007

1 Department of Obstetrics and Gynecology, Kobe University Graduate School of Medicine, Kobe, Japan; and 2 TAP Pharmaceutical Products, Lake Forest, Illinois Submitted 4 April 2007 ; accepted in final form 15 July 2007 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 2 , 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. growth-arrest- and DNA-damage-inducible gene 153; Bcl-2 family; tribbles-related protein-3; GADD34 Address for reprint requests and other correspondence: T. Maruo, Dept. of Obstetrics and Gynecology, Kobe Univ. Graduate School of Medicine, 7-5-1 Kusunoki-C