Downregulation of retinoblastoma protein is involved in the enhanced cytotoxicity of 4-hydroxytamoxifen plus mifepristone combination therapy versus antiestrogen monotherapy of human breast cancer.
In this study, human MCF-7 breast cancer cells, which express functional estrogen and progesterone receptors, were used to compare the efficacy of combined antiestrogen plus antiprogestin therapy to antiestrogen monotherapy. Cells were treated with the antiestrogen 4-hydroxytamoxifen (4-OHT) and/or the antiprogestin mifepristone (MIF) and effects on cell proliferation (cytostatic action), cell cycle phase, the phosphorylation state of the tumor suppressor retinoblastoma protein (Rb), and induction of active cell death (cytotoxic action) were determined. Combination hormonal therapy showed both increased cytostatic and cytotoxic activity as compared to either monotherapy. The increased cytostatic action was mediated by Rb activation; whereas, the cytotoxic (pro-apoptotic) action of combined hormonal therapy correlated to a significant reduction in Rb protein levels. To test the apparent role of Rb protein loss in the pro-apoptotic action of combined hormonal therapy, Rb was downregulated in MCF-7 cells using siRNA-targeting. The siRNA-mediated knockdown of Rb combined with 4-OHT therapy resulted in a pro-apoptotic action similar to that resulting from 4-OHT and MIF combination treatment, which included increased cell detachment from the monolayer, high-molecular-weight genomic DNA fragmentation, and cleavage of poly ADP-ribose polymerase (PARP) and lamin A. From these studies, we conclude that Rb protein downregulation is required for 4-OHT-treated, estrogen receptor positive (ER+) breast cancer cells to undergo active cell death. We discuss the potential of using an antiprogestin such as MIF plus antiestrogen treatment to more effectively downregulate Rb in ER+ breast cancer cells to increase the overall cytotoxic action of hormonal therapy.
Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta, GA 30912-2000, USA. pschoenl@mail.mcg.edu
Patricia V
PV
Schoenlein
Min
M
Hou
Julia S
JS
Samaddar
Virgil T
VT
Gaddy
Muthusamy
M
Thangaraju
Jill
J
Lewis
Maribeth
M
Johnson
Vadivel
V
Ganapathy
Andre
A
Kallab
John T
JT
Barrett
D000971:Antineoplastic Combined Chemotherapy Protocols / Q000627:therapeutic use*; D001943:Breast Neoplasms / Q000188:drug therapy*; D016923:Cell Death; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D053938:DNA Fragmentation; D015536:Down-Regulation; D004965:Estrogen Antagonists / Q000494:pharmacology*; D004967:Estrogens / Q000737:chemistry; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D015735:Mifepristone / Q000008:administration & dosage*; D034741:RNA, Small Interfering / Q000378:metabolism; D016160:Retinoblastoma Protein / Q000378:metabolism; D013629:Tamoxifen / Q000008:administration & dosage / Q000031:analogs & derivatives*
D004965:Estrogen Antagonists; D004967:Estrogens; D034741:RNA, Small Interfering; D016160:Retinoblastoma Protein; D013629:Tamoxifen; C016601:afimoxifene; D015735:Mifepristone