Acteoside inhibits human promyelocytic HL-60 leukemia cell proliferation via inducing cell cycle arrest at G0/G1 phase and differentiation into monocyte.
We investigated the in vitro effects of acteoside on the proliferation, cell cycle regulation and differentiation of HL-60 human promyelocytic leukemia cells. Acteoside inhibited the proliferation of HL-60 cells in a concentration- and time-dependent manner with an IC50, approximately 30 microM. DNA flow cytometric analysis indicated that acteoside blocked cell cycle progression at the G1 phase in HL-60 human promyelocytic leukemia cells. Among the G1 phase cell cycle-related proteins, the levels of cyclin-dependent protein kinase (CDK)2, CDK6, cyclin D1, cyclin D2, cyclin D3 and cyclin E were reduced by acteoside, whereas the steady-state level of CDK4 was unaffected. The protein and mRNA levels of CDK inhibitors (cyclin-dependent kinase inhibitors), such as p21(CIP1/WAF1) and p27(KIP1), were gradually increased after acteoside treatment in a time-dependent manner. In addition, acteoside markedly enhanced the binding of p21(CIP1/WAF1) and p27(KIP1) to CDK4 and CDK6, resulting in the reduction of CDK2, CDK4 and CDK6 activities. Moreover, the hypophosphorylated form of retinoblastoma increased, leading to the enhanced binding of protein retinoblastoma (pRb) and E2F1. Our results further suggest that acteoside is a potent inducer of differentiation of HL-60 cells based on biochemical activities and the expression level of CD14 cell surface antigen. In conclusion, the onset of acteoside-induced G1 arrest of HL-60 cells prior to the differentiation appears to be tightly linked to up-regulation of the p21(CIP1/WAF1) and p27(KIP1) levels and decreases in the CDK2, CDK4 and CDK6 activities. These findings, for the first time, reveal the mechanism underlying the anti-proliferative effect of acteoside on human promyelocytic HL-60 cells.
Lee
Kyung-Won
KW
Department of Pharmaceutical Biochemistry, College of Pharmacy, Kyung-Hee University, Hoegi-Dong, Seoul 130-701, Korea.
Kim
Hyoung Ja
HJ
Lee
Yong Sup
YS
Park
Hee-Jun
HJ
Choi
Jong-Won
JW
Ha
Joohun
J
Lee
Kyung-Tae
KT
D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects*; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D004815:Epidermal Growth Factor / Q000494:pharmacology; D016193:G1 Phase / Q000187:drug effects; D005960:Glucosides / Q000494:pharmacology*; D018922:HL-60 Cells / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D032342:Muscle Cells / Q000166:cytology / Q000187:drug effects; D010636:Phenols / Q000494:pharmacology*; D016192:Resting Phase, Cell Cycle / Q000187:drug effects; D000077185:Resveratrol; D013267:Stilbenes / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D059808:Polyphenols
D000972:Antineoplastic Agents, Phytogenic; D004815:Epidermal Growth Factor; D005960:Glucosides; D010636:Phenols; D000077185:Resveratrol; D013267:Stilbenes; D013755:Tetradecanoylphorbol Acetate; C058956:acteoside; D059808:Polyphenols