Cellular thiol status-dependent inhibition of tumor cell growth via modulation of p27(kip1) translocation and retinoblastoma protein phosphorylation by 1'-acetoxychavicol acetate.
1'-Acetoxychavicol acetate (ACA) has been shown to inhibit tumor cell growth, but there is limited information on its effects on cell signaling and the cell cycle control pathway. In this study, we sought to determine how ACA alters cell cycle and its related control factors in its growth inhibitory effect in Ehrlich ascites tumor cells (EATC). ACA caused an accumulation of cells in the G1 phase and an inhibition of DNA synthesis, which were reversed by supplementation with N-acetylcysteine (NAC) or glutathione ethyl ester (GEE). Furthermore, ACA decreased hyperphosphorylated Rb levels and increased hypophosphorylated Rb levels. NAC and GEE also abolished the decease in Rb phosphorylation by ACA. As Rb phosphorylation is regulated by G1 cyclin dependent kinase and CDK inhibitor p27(kip1), which is an important regulator of the mammalian cell cycle, we estimated the amount of p27(kip1) levels by western blotting. Treatment with ACA had virtually no effect on the amount of p27(kip1) levels, but caused a decrease in phosphorylated p27(kip1) and an increase in unphosphorylated p27(kip1) as well as an increase in the nuclear localization of p27(kip1). These events were abolished in the presence of NAC or GEE. These results suggest that in EATC, cell growth inhibition elicited by ACA involves decreases in Rb and p27(kip1) phosphorylation and an increase in nuclear localization of p27(kip1), and these events are dependent on the cellular thiol status.
Unahara
Y
Y
Department of Food and Human Health Sciences, Graduate School of Human Life Science, Osaka City University, Osaka, Japan.
Kojima-Yuasa
A
A
Higashida
M
M
Kennedy
D O
DO
Murakami
A
A
Ohigashi
H
H
Matsui-Yuasa
I
I
D000111:Acetylcysteine / Q000378:metabolism; D000818:Animals; D001592:Benzyl Alcohols; D002286:Carcinoma, Ehrlich Tumor; D002453:Cell Cycle / Q000502:physiology*; D045744:Cell Line, Tumor; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000378:metabolism*; D004247:DNA / Q000096:biosynthesis; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D010766:Phosphorylation; D010936:Plant Extracts / Q000737:chemistry / Q000378:metabolism; D016160:Retinoblastoma Protein / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000737:chemistry / Q000378:metabolism*; D013729:Terpenes / Q000737:chemistry / Q000378:metabolism*
D001592:Benzyl Alcohols; D010936:Plant Extracts; D016160:Retinoblastoma Protein; D013438:Sulfhydryl Compounds; D013729:Terpenes; D050760:Cyclin-Dependent Kinase Inhibitor p27; C042431:S-ethyl glutathione; D004247:DNA; D005978:Glutathione; C047948:1'-acetoxychavicol acetate; D000111:Acetylcysteine