유방암 세포주(T47D와 MDA-MB231)에서 Genistein의 세포주기 G2/M 정지 기전 = Molecular Mechanism of the G2/M Arrest in Breast Cancer Cells (T47D and MDA-MB231) Induced by Genistein

다국어 초록 (Multilingual Abstract)

Purpose: To analyze the effect of the growth control on human breast cancer cells with genistein treatment and to investigate the mechanism of genistein-induced G2/M arrest in T47D and MDA-MB231 breast carcinoma cells by Cdc25C expression. Methods: We analysed the proliferartion of the two cell lines by using MTT proliferation assay, flow cytometric analysis, real-time quantitative RT-PCR and western blotting and investigated the effect of genistein on cell survival, cellular toxicity, cell cycle progression-related genes and their mRNA and protein alterations. Results: The DNA flow cytometric analysis of both cell lines treated with genistein showed a dose-dependent growth inhibition and accumulation in the G2/M phase of cell cycle. The expression of p21 mRNA and protein increased in both cell lines following genistein treatment but p27 expression was unchanged. Furthermore,decreased Cdc25C expression with decreased polo-like kinase (PLK) 1 expression and increased PLK3 expression were observed after genistein treatment. The decreased level of Cdc25C in the nucleus was associated with decreased phosphorylation of Cdc25C by PLK1. The expression of PLK3was increased with a dose-dependent and a time-dependent manner and was associated with decreased Cdc25C expression. Check point kinase (CHK) 1 and CHK2 revealed different expression patterns each other. The CHK1 expression was independent of the presence of genestein. CHK2 expression increased in MDA-MB231 cells associated with decreased Cdc25C expression but not in T47D. Conclusion: These results suggest that genistein induces a G2/M arrest in human breast cancer cells, the mechanism of which is due, in part, to decreased in Cdc25C phosphatase by a regulatory effect of PLK1, PLK3, and CHK2 as well as increased expression of the cyclin dependent kinase inhibitor p21(WAF1/CIP1).

참고문헌 (Reference)

1 Casagrande F, "p21CIP1 is dispensable for the G2 arrest caused by genistein in human melanoma cells" 258 : 101-108, 2000

2 Mueller SC, "Tyrosine phosphorylation of membrane proteins mediates cellular invasion by transformed cells" 119 : 1309-1325, 1992

3 Boutros R, "The when and wheres of CDC25 phosphatases" 18 : 185-191, 2006

4 Myer DL, "The Plk3-Cdc25 circuit" 24 : 299-305, 2005

5 Hunter T, "Protein-tyrosine kinases" 54 : 897-930, 1985

6 Bahassi el M, "Priming phosphorylation of Chk2 by polo-like kinase 3 (Plk3) mediates its full activation by ATM and a downstream checkpoint in response to DNA damage" 596 : 166-176, 2006

7 van de Weerdt BC, "Polo-like kinases: a team in control of the division" 5 : 853-864, 2006

8 Eckerdt F, "Polo-like kinases and oncogenesis" 24 : 267-276, 2005

9 Jiang N, "Polo box domain of Plk3 functions as a centrosome localization signal, overexpression of which causes mitotic arrest, cytokinesis defects, and apoptosis" 281 : 10577-10582, 2006

10 Xie S, "Plk3 functionally links DNA damage to cell cycle arrest and apoptosis at least in part via the p53 pathway" 276 : 43305-43312, 2001

11 Waite KA, "Phytoestrogen exposure elevates PTEN levels" 14 : 1457-1463, 2005

12 Ding H, "P21 response to DNA damage induced by genistein and etoposide in human lung cancer cells" 305 : 950-956, 2003

13 Linassier C, "Mechanisms of action in NIH-3T3 cells of genistein, an inhibitor of EGF receptor tyrosine kinase activity" 39 : 187-193, 1990

14 Markovits J, "Inhibitory effects of the tyrosine kinase inhibitor genistein on mammalian DNA topoisomerase II" 49 : 5111-5117, 1989

15 Zhu H, "Identification of promoter elements responsible for transcriptional inhibition of polo-like kinase 1 and topoisomerase IIalpha genes by p21(WAF1/CIP1/SDI1)" 1 : 59-66, 2002

16 Ismail IA, "Genistein-induced neuronal apoptosis and G2/M cell cycle arrest is associated with MDC1 up-regulation and PLK1 down-regulation" 575 : 12-20, 2007

17 Choi YH, "Genistein-induced G2/M arrest is associated with the inhibition of cyclin B1 and the induction of p21 in human breast carcinoma cells" 13 : 391-396, 1998

18 Akiyama T, "Genistein, a specific inhibitor of tyrosine-specific protein kinases" 262 : 5592-5595, 1987

19 Fotsis T, "Genistein, a dietary-derived inhibitor of in vitro angiogenesis" 90 : 2690-2694, 1993

20 Shao ZM, "Genistein inhibits proliferation similarly in estrogen receptor-positive and negative human breast carcinoma cell lines characterized by P21WAF1/CIP1 induction, G2/M arrest, and apoptosis" 69 : 44-54, 1998

21 Kuzumaki T, "Genistein induces p21(Cip1/WAF1) expression and blocks the G1 to S phase transition in mouse fibroblast and melanoma cells" 251 : 291-295, 1998

22 Cappelletti V, "Genistein blocks breast cancer cells in the G(2)M phase of the cell cycle" 79 : 594-600, 2000

23 Chang KL, "Genistein arrests hepatoma cells at G2/M phase: involvement of ATM activation and upregulation of p21waf1/cip1 and Wee1" 67 : 717-726, 2004

24 Frey RS, "Genistein activates p38 mitogen-activated protein kinase, inactivates ERK1/ERK2 and decreases Cdc25C expression in immortalized human mammary epithelial cells" 133 : 226-231, 2003

25 Frey RS, "Effects of genistein on cell proliferation and cell cycle arrest in nonneoplastic human mammary epithelial cells: involvement of Cdc2, p21(waf/cip1), p27(kip1), and Cdc25C expression" 61 : 979-989, 2001

26 Upadhyay S, "Differential sensitivity of normal and malignant breast epithelial cells to genistein is partly mediated by p21(WAF1)" 7 : 1782-1789, 2001

27 Yoon DS, "Different expression of p27(kip1) and p57(kip2) by genistein treatment in MDA-MB-231 human breast cancer cells" 7 : 98-103, 2004

28 Agarwal R, "Cell signaling and regulators of cell cycle as molecular targets for prostate cancer prevention by dietary agents" 60 : 1051-1059, 2000

29 Masuda Y, "Beta-hydroxyisovalerylshikonin induces apoptosis in human leukemia cells by inhibiting the activity of a polo-like kinase 1 (PLK1)" 22 : 1012-1023, 2003

30 Livak KJ, "Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method" 25 : 402-408, 2001