Role of extracellular matrix in regulation of staurosporine-induced apoptosis in breast cancer cells.
Autocrine and paracrine mechanisms modulate the synthesis and secretion of extracellular matrix (ECM); moreover, each component of the ECM is capable of modulating the synthesis and release of other ECM molecules. Therefore, the synthesis of ECM glycoprotein fibronectin and laminin was studied in the human breast cancer cell lines MCF7 and MDA MB 23, plated on different ECM. Our results showed that the cells plated on a fibronectin substrate increased laminin synthesis: this event correlated with an increase in alpha2 and alpha3 integrin subunits. Staurosporine-induced apoptosis was then analyzed in the cell lines plated on different ECM. Staurosporine treatment determined the apoptosis of 35 and 33% respectively of MDA MB 231 and MCF7; these values increased to 60 and 64% in cells plated on laminin, to 48 and 63% in cells plated on fibronectin and to 64 and 69% in cells plated on matrigel. Moreover, staurosporine treatment decreased bcl-2 expression in the cells plated on fibronectin and laminin. Yet, staurosporine treatment determined PARP cleavage and PARP partial disappearance when the cells were plated on matrigel. Finally, a partial loss of function mutant Ras protein that activated only Raf pathway, was expressed in MCF7, in order to identify whether the increase of apoptosis induced by extracellular matrix involved the Raf/MAP kinase pathway. The increase of apoptosis of the cells plated on matrigel suggested that the activation of the Raf pathway is probably involved in the decrease of survival on matrigel. These data demonstrate that the modification of ECM modulates the apoptotic process of breast cancer cells and suggest that it is worthwhile to dissect the role of ECM in the control of apoptotic process.
Vasaturo
F
F
Department of Experimental Medicine and Pathology, University of Rome La Sapienza, viale Regina Elena 324, 00161 Rome, Italy.
Malacrino
C
C
Sallusti
E
E
Coppotelli
G
G
Birarelli
P
P
Giuffrida
A
A
Albonici
L
L
Simonelli
L
L
Modesti
A
A
Modesti
M
M
Scarpa
S
S
D017209:Apoptosis; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D016923:Cell Death; D045744:Cell Line, Tumor; D002469:Cell Separation; D003094:Collagen / Q000494:pharmacology; D004338:Drug Combinations; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005109:Extracellular Matrix / Q000378:metabolism*; D005353:Fibronectins / Q000096:biosynthesis / Q000737:chemistry; D005434:Flow Cytometry; D006801:Humans; D047468:Immunoprecipitation; D007797:Laminin / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D011107:Polylysine / Q000737:chemistry; D011509:Proteoglycans / Q000494:pharmacology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D019311:Staurosporine / Q000494:pharmacology*; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D051028:bcl-2-Associated X Protein
D004338:Drug Combinations; D004791:Enzyme Inhibitors; D005353:Fibronectins; D007797:Laminin; D011509:Proteoglycans; D019253:Proto-Oncogene Proteins c-bcl-2; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein; C059485:matrigel; D011107:Polylysine; D003094:Collagen; D011065:Poly(ADP-ribose) Polymerases; D019311:Staurosporine