Withaferin A causes FOXO3a- and Bim-dependent apoptosis and inhibits growth of human breast cancer cells in vivo.
Withaferin A (WA) is derived from the medicinal plant Withania somnifera, which has been safely used for centuries in Indian Ayurvedic medicine for treatment of different ailments. We now show, for the first time, that WA exhibits significant activity against human breast cancer cells in culture and in vivo. The WA treatment decreased viability of MCF-7 (estrogen-responsive) and MDA-MB-231 (estrogen-independent) human breast cancer cells in a concentration-dependent manner. The WA-mediated suppression of breast cancer cell viability correlated with apoptosis induction characterized by DNA condensation, cytoplasmic histone-associated DNA fragmentation, and cleavage of poly-(ADP-ribose)-polymerase. On the other hand, a spontaneously immortalized normal mammary epithelial cell line (MCF-10A) was relatively more resistant to WA-induced apoptosis compared with breast cancer cells. The WA-mediated apoptosis was accompanied by induction of Bim-s and Bim-L in MCF-7 cells and induction of Bim-s and Bim-EL isoforms in MDA-MB-231 cells. The cytoplasmic histone-associated DNA fragmentation resulting from WA exposure was significantly attenuated by knockdown of protein levels of Bim and its transcriptional regulator FOXO3a in both cell lines. Moreover, FOXO3a knockdown conferred marked protection against WA-mediated induction of Bim-s expression. The growth of MDA-MB-231 cells implanted in female nude mice was significantly retarded by 5 weekly i.p. injections of 4 mg WA/kg body weight. The tumors from WA-treated mice exhibited reduced cell proliferation and increased apoptosis compared with tumors from control mice. These results point toward an important role of FOXO3a and Bim in regulation of WA-mediated apoptosis in human breast cancer cells.
Stan
Silvia D
SD
Department of Pharmacology and Chemical Biology, and University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
Hahm
Eun-Ryeong
ER
Warin
Renaud
R
Singh
Shivendra V
SV
D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism*; D000072224:Bcl-2-Like Protein 11; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D048708:Cell Growth Processes / Q000187:drug effects; D045744:Cell Line, Tumor; D004875:Ergosterol / Q000031:analogs & derivatives* / Q000494:pharmacology; D005260:Female; D000071316:Forkhead Box Protein O3; D051858:Forkhead Transcription Factors / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008819:Mice, Nude; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection; D054358:Withanolides; D023041:Xenograft Model Antitumor Assays
D000972:Antineoplastic Agents, Phytogenic; D051017:Apoptosis Regulatory Proteins; C000606775:BCL2L11 protein, human; D000072224:Bcl-2-Like Protein 11; C000606776:Bcl2l11 protein, mouse; C499517:FOXO3 protein, human; D000071316:Forkhead Box Protein O3; D051858:Forkhead Transcription Factors; D008565:Membrane Proteins; D011518:Proto-Oncogene Proteins; D034741:RNA, Small Interfering; D054358:Withanolides; C009684:withaferin A; D004875:Ergosterol
Science. 1998 Aug 28;281(5381):1322-6
9735050
Carcinogenesis. 2003 May;24(5):891-7
12771033
Curr Biol. 2000 Oct 5;10(19):1201-4
11050388
Neuron. 2001 Mar;29(3):615-28
11301022
Cancer Lett. 1995 Aug 16;95(1-2):189-93
7656229
Lancet. 1995 Sep 30;346(8979):883-7
7564675
Mol Cancer Ther. 2006 Nov;5(11):2931-45
17121941
Indian J Exp Biol. 2000 May;38(5):432-7
11272405
CA Cancer J Clin. 2006 Mar-Apr;56(2):106-30
16514137
J Nat Prod. 2003 Jul;66(7):1022-37
12880330
Cancer Res. 2007 Jan 1;67(1):246-53
17185378
Vascul Pharmacol. 2006 Jun;44(6):406-10
16713367
J Ethnopharmacol. 1999 Oct;67(1):27-35
10616957
Clin Cancer Res. 2008 Feb 15;14(4):1248-57
18281560
Hum Exp Toxicol. 2005 Mar;24(3):137-47
15901053
Cancer Res. 2005 Mar 1;65(5):2035-43
15753404
Mol Cancer Ther. 2005 Oct;4(10):1495-504
16227398
Mol Cancer Ther. 2006 Jun;5(6):1434-45
16818501
J Biol Chem. 2001 Oct 12;276(41):37754-60
11495903
Mol Pharmacol. 2007 Feb;71(2):426-37
17093135
Br J Cancer. 2007 Jan 15;96(1):110-7
17164758
J Biol Chem. 2008 Feb 1;283(5):2508-17
18063582
Neuron. 2003 Jun 19;38(6):899-914
12818176
Phytomedicine. 2005 Mar;12(3):229-35
15830846
Altern Med Rev. 2000 Aug;5(4):334-46
10956379
Neuron. 2001 Mar;29(3):629-43
11301023
Annu Rev Public Health. 1996;17:47-67
8724215
Apoptosis. 2007 Nov;12(11):2115-33
17874299
J Biol Chem. 2007 Feb 16;282(7):4253-4264
17150968
J Natl Cancer Inst. 1998 Sep 16;90(18):1371-88
9747868
Indian J Exp Biol. 1992 Mar;30(3):169-72
1512021
J Cell Physiol. 2000 Feb;182(2):171-81
10623880
Cancer Res. 2007 Aug 1;67(15):7439-49
17671214
Annu Rev Immunol. 1998;16:395-419
9597135
Lancet. 2002 Sep 14;360(9336):817-24
12243915
Phytother Res. 2007 Oct;21(10):905-13
17562568
Clin Cancer Res. 2006 Nov 15;12(22):6836-43
17121905
Carcinogenesis. 2004 Jan;25(1):83-90
14514658
Mol Cell. 1999 Mar;3(3):287-96
10198631
Am J Pathol. 2000 Aug;157(2):449-61
10934149
Oncogene. 2004 Jul 22;23(33):5594-606
15184882
Epidemiol Rev. 1993;15(1):36-47
8405211
Int J Radiat Biol. 1996 Feb;69(2):193-7
8609455
Angiogenesis. 2004;7(2):115-22
15516832
Nat Chem Biol. 2006 Jan;2(1):33-8
16408090
Mol Cell Biochem. 2004 May;260(1-2):39-47
15228084