Progesterone receptor modulator CDB-2914 down-regulates proliferative cell nuclear antigen and Bcl-2 protein expression and up-regulates caspase-3 and poly(adenosine 5'-diphosphate-ribose) polymerase expression in cultured human uterine leiomyoma cells.

Abstract

The present study was conducted to evaluate the effects of the progesterone receptor modulator CDB-2914 on proliferative activity and apoptosis in cultured human uterine leiomyoma cells. Isolated leiomyoma cells were subcultured in phenol red-free DMEM supplemented with 10% fetal bovine serum for 120 h and then stepped down to serum-free conditions for 12, 24, 48, and 96 h in the absence or presence of graded concentrations of CDB-2914 (10(-9), 10(-8), 10(-7), and 10(-6) M). The number of viable cultured leiomyoma cells was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazodium bromide assay. Proliferating cell nuclear antigen (PCNA) expression was evaluated by immunocytochemistry and Western blot analysis. Apoptosis was examined by terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate nick end labeling (TUNEL) assay. Caspase-3, cleaved poly(ADP-ribose) polymerase (PARP), and Bcl-2 expression were assessed by Western blot analysis. Compared with untreated control cultures, treatment with CDB-2914 decreased the number of viable cultured leiomyoma cells and the PCNA-positive rate in those cells and increased the TUNEL-positive rate in cultured leiomyoma cells in a dose-dependent manner. Western blot analysis revealed that treatment with CDB-2914 significantly decreased the expression of PCNA and Bcl-2 protein and increased the expression of cleaved caspase-3 and cleaved PARP in a dose-dependent manner compared with untreated control cultures. These results suggest that CDB-2914 inhibits the proliferation of cultured leiomyoma cells by down-regulating PCNA expression and induces apoptosis by up-regulating cleaved caspase-3 and PARP expression and down-regulating Bcl-2 protein expression in those cells.

• Authors: Qin Xu, Shigeki Takekida, Noriyuki Ohara, Wei Chen, Regine Sitruk-Ware, Elof D B Johansson, Takeshi Maruo
• Journal: The Journal of clinical endocrinology and metabolism (J Clin Endocrinol Metab) Vol. 90 Issue 2 Pg. 953-61 (Feb 2005) ISSN: 0021-972X [Print] United States
• PMID: 15572421 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Norpregnadienes
• Proliferating Cell Nuclear Antigen
• Proto-Oncogene Proteins c-bcl-2
• Receptors, Progesterone
• ulipristal
• Poly(ADP-ribose) Polymerases
• CASP3 protein, human
• Caspase 3
• Caspases

Topics

• Apoptosis (drug effects)
• Blotting, Western
• Caspase 3
• Caspases (metabolism)
• Female
• Gene Expression Regulation, Enzymologic (drug effects)
• Gene Expression Regulation, Neoplastic (drug effects)
• Humans
• Immunohistochemistry
• Leiomyoma (pathology)
• Norpregnadienes (pharmacology)
• Poly(ADP-ribose) Polymerases (metabolism)
• Proliferating Cell Nuclear Antigen (drug effects, genetics)
• Proto-Oncogene Proteins c-bcl-2 (metabolism)
• Receptors, Progesterone (drug effects, physiology)
• Tumor Cells, Cultured
• Uterine Neoplasms (pathology)