Knock-Down of Amphiregulin Inhibits Cellular Invasion in Inflammatory Breast Cancer

被引:37
作者
Baillo, Andrea [1 ,2 ]
Giroux, Craig [1 ,2 ]
Ethier, Stephen P. [1 ,2 ]
机构
[1] Wayne State Univ, Dept Oncol, Detroit, MI USA
[2] Karmanos Canc Inst, Detroit, MI USA
关键词
EPIDERMAL-GROWTH-FACTOR; MAMMARY-GLAND DEVELOPMENT; FACTOR RECEPTOR EGFR; EPITHELIAL-CELLS; LUNG-CANCER; IN-VITRO; EXPRESSION; AUTOCRINE; CARCINOMA; PROLIFERATION;
D O I
10.1002/jcp.22620
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
We have previously shown that SUM-149 human breast cancer cells require an amphiregulin (AREG) autocrine loop for cell proliferation. We also demonstrated that AREG can increase epidermal growth factor receptor (EGFR) stability and promote EGFR localization to the plasma membrane. In the present studies we successfully knocked-down AREG expression in SUM-149 cells by lentiviral infection of AREG shRNA. In the absence of AREG expression, SUM-149 cell growth was slowed, but not completely inhibited. Furthermore, cells infected with AREG shRNA constructs showed an increase in EGFR protein expression by Western blot. Immunofluorescence and confocal microscopy showed that following AREG knock-down, EGFR continued to localize to the cell surface. Soft agar assays demonstrated that AREG knock-down cells retain anchorage-independent growth capacity. Additionally mammosphere forming assays and Adefluor staining analysis showed that knock-down of AREG expression did not affect the expression of stem cell phenotypes. However, following AREG knock-down, SUM-149 cells demonstrated a dramatic decrease in their ability to invade a Matrigel matrix. Consistent with this observation, microarray analysis comparing cells infected with a non-silencing vector to the AREG knock-down cells, identified genes associated with the invasive phenotype such as RHOB and DKK1, and networks associated with cell motility such as integrin-linked kinase signaling, and focal adhesion kinase signaling. AREG was also found to modulate WNT and Notch signaling in these cells. Thus, AREG functions in regulating the invasive phenotype, and we propose that this regulation may be through altered signaling that occurs when AREG activates plasma membrane localized EGFR. J. Cell. Physiol. J. Cell. Physiol. 226: 2691-2701, 2011. (C) 2011 Wiley-Liss, Inc.
引用
收藏
页码:2691 / 2701
页数:11
相关论文
共 52 条
[31]   Epidermal growth factor (EGF) increases the in vitro invasion, motility and adhesion interactions of the primary renal carcinoma cell line, A704 [J].
Price, JT ;
Wilson, HM ;
Haites, NE .
EUROPEAN JOURNAL OF CANCER, 1996, 32A (11) :1977-1982
[32]   Wnt 5a signaling is critical for macrophage-induced invasion of breast cancer cell lines [J].
Pukrop, T ;
Klemm, F ;
Hagemann, T ;
Gradl, D ;
Schulz, M ;
Siemes, S ;
Trümper, L ;
Binder, C .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2006, 103 (14) :5454-5459
[33]   EPIDERMAL GROWTH FACTOR-RELATED PEPTIDES AND THEIR RECEPTORS IN HUMAN MALIGNANCIES [J].
SALOMON, DS ;
BRANDT, R ;
CIARDIELLO, F ;
NORMANNO, N .
CRITICAL REVIEWS IN ONCOLOGY HEMATOLOGY, 1995, 19 (03) :183-232
[34]   THE ROLE OF AMPHIREGULIN IN BREAST-CANCER [J].
SALOMON, DS ;
NORMANNO, N ;
CIARDIELLO, F ;
BRANDT, R ;
SHOYAB, M ;
TODARO, GJ .
BREAST CANCER RESEARCH AND TREATMENT, 1995, 33 (02) :103-114
[35]  
SCHLEGEL J, 1994, INT J CANCER, V56, P72
[36]  
Schuger L, 1996, DEVELOPMENT, V122, P1759
[37]   EPIDERMAL GROWTH-FACTOR RECEPTOR-DEPENDENT STIMULATION OF AMPHIREGULIN EXPRESSION IN ANDROGEN-STIMULATED HUMAN PROSTATE-CANCER CELLS [J].
SEHGAL, I ;
BAILEY, J ;
HITZEMANN, K ;
PITTELKOW, MR ;
MAIHLE, NJ .
MOLECULAR BIOLOGY OF THE CELL, 1994, 5 (03) :339-347
[38]   STRUCTURE AND FUNCTION OF HUMAN AMPHIREGULIN - A MEMBER OF THE EPIDERMAL GROWTH-FACTOR FAMILY [J].
SHOYAB, M ;
PLOWMAN, GD ;
MCDONALD, VL ;
BRADLEY, JG ;
TODARO, GJ .
SCIENCE, 1989, 243 (4894) :1074-1076
[39]   AMPHIREGULIN - A BIFUNCTIONAL GROWTH-MODULATING GLYCOPROTEIN PRODUCED BY THE PHORBOL 12-MYRISTATE 13-ACETATE-TREATED HUMAN-BREAST ADENOCARCINOMA CELL-LINE MCF-7 [J].
SHOYAB, M ;
MCDONALD, VL ;
BRADLEY, JG ;
TODARO, GJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (17) :6528-6532
[40]  
TOI M, 1990, CANCER-AM CANCER SOC, V65, P1980, DOI 10.1002/1097-0142(19900501)65:9<1980::AID-CNCR2820650917>3.0.CO