Silencing of the HER2/neu gene by siRNA inhibits proliferation and induces apoptosis in HER2/neu-overexpressing breast cancer cells.
In eukaryotes, double-stranded (ds) RNA induces sequence-specific inhibition of gene expression referred to as RNA interference (RNAi). We exploited RNAi to define the role of HER2/neu in the neoplastic proliferation of human breast cancer cells. We transfected SK-BR-3, BT-474, MCF-7, and MDA-MB-468 breast cancer cells with short interfering RNA (siRNA) targeted against human HER2/neu and analyzed the specific inhibition of HER2/neu expression by Northern and Western blots. Transfection with HER2/neu-specific siRNA resulted in a sequence-specific decrease in HER2/neu mRNA and protein levels. Moreover, transfection with HER2/neu siRNA caused cell cycle arrest at G0/G1 in the breast cancer cell lines SK-BR-3 and BT-474, consistent with a powerful RNA silencing effect. siRNA treatment resulted in an antiproliferative and apoptotic response in cells overexpressing HER2/neu, but had no influence in cells with almost no expression of HER2/neu proteins like MDA-MB-468 cells. These data indicate that HER2/neu function is essential for the proliferation of HER2/neu-overexpressing breast cancer cells. Our observations suggest that siRNA targeted against human HER2/neu may be valuable tools as antiproliferative agents that display activity against neoplastic cells at very low doses.
Faltus
Timo
T
Department of Obstetrics and Gynecology, Medical School, J. W. Goethe University, Theodor-Stern-Kai 7, Frankfurt 60590, Germany.
Yuan
Juping
J
Zimmer
Brigitte
B
Krämer
Andrea
A
Loibl
Sibylle
S
Kaufmann
Manfred
M
Strebhardt
Klaus
K
D017209:Apoptosis / Q000502:physiology*; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000235:genetics*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D020868:Gene Silencing; D018734:Genes, erbB-2 / Q000235:genetics*; D006801:Humans; D034741:RNA, Small Interfering / Q000378:metabolism*; D014162:Transfection
D034741:RNA, Small Interfering
J Clin Oncol. 1996 Mar;14(3):737-44
8622019
Science. 1987 Jan 9;235(4785):177-82
3798106
Int J Cancer. 2004 Jan 1;108(1):71-7
14618618
Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8691-5
1717984
Nat Med. 1995 Nov;1(11):1119-21
7584974
Nature. 1984 Dec 6-12;312(5994):545-8
6504162
Int J Cancer. 1997 Dec 10;73(6):875-9
9399669
J Clin Oncol. 1989 Aug;7(8):1120-8
2569032
Nature. 1984 May 31-Jun 6;309(5967):418-25
6328312
Nature. 2002 Jul 11;418(6894):244-51
12110901
Biochem Biophys Res Commun. 1994 Apr 15;200(1):661-7
7909438
Oncogene. 1997 Feb 6;14(5):543-9
9053852
Mol Cell Biol. 1995 Mar;15(3):1182-91
7532277
Cancer Res. 2000 Feb 1;60(3):560-5
10676637
EMBO J. 1999 Oct 15;18(20):5528-39
10523297
Nat Med. 2000 Apr;6(4):443-6
10742152
Am J Obstet Gynecol. 1991 Jan;164(1 Pt 1):15-21
1670908
Int J Cancer. 1989 Feb 15;43(2):270-2
2563720
Nature. 2000 Mar 16;404(6775):293-6
10749213
Br J Cancer. 1994 Nov;70(5):819-25
7947086
Cancer Cell. 2002 Mar;1(2):117-23
12086869
Oncogene. 2000 Dec 11;19(53):6138-43
11156527
Nat Biotechnol. 2003 Jun;21(6):635-7
12754523
Breast Cancer Res Treat. 1992;24(2):85-95
8095168
Nature. 2001 May 24;411(6836):494-8
11373684
Nat Med. 2003 Mar;9(3):347-51
12579197
J Biol Chem. 2004 Feb 6;279(6):4339-45
14625284
Mol Cell Biol. 2000 May;20(9):3210-23
10757805
Oncogene. 2003 Dec 8;22(56):9087-96
14663487
J Clin Oncol. 1998 Aug;16(8):2659-71
9704716
Cell. 2001 Nov 2;107(3):309-21
11701122
Cancer Res. 1990 Jul 1;50(13):4087-91
1972347
Mol Cell Biol. 1989 Mar;9(3):1165-72
2566907
J Surg Res. 1998 Jun;77(1):85-90
9698539
Cell. 2000 Mar 31;101(1):25-33
10778853
Nat Rev Genet. 2002 Oct;3(10):737-47
12360232
Proc Natl Acad Sci U S A. 1992 May 15;89(10):4285-9
1350088
J Natl Cancer Inst. 2002 Dec 18;94(24):1863-77
12488480
Mol Cell. 2000 Nov;6(5):1077-87
11106747
J Biol Chem. 1996 Oct 4;271(40):24747-52
8798744
Oncogene. 2002 Nov 28;21(54):8282-92
12447691
Oncogene. 2002 Aug 22;21(37):5716-24
12173041
Proc Natl Acad Sci U S A. 2003 May 13;100(10):5789-94
12732729