BACKGROUND: Proliferative breast disease (PBD) may increase a woman's risk of developing breast cancer, perhaps by decreasing cellular sensitivity to apoptosis. To determine whether resistance to apoptosis develops during PBD, we investigated apoptosis initiated through the Fas pathway in a series of cell lines that recapitulates the morphologic changes of PBD in nude/beige mice. METHODS: The series of cell lines used was MCF-10A cells (parental preneoplastic human breast epithelial cells), MCF-10AT cells (transformed with T(24) Ha-ras), and MCF-10ATG3B cells (derivative cells that progress to carcinoma). Fas-mediated apoptosis, induced when a Fas monoclonal antibody bound to and activated the Fas receptor on these cells, was assessed morphologically and by flow cytometry. Levels of proteins involved in Fas-mediated apoptosis and cleavage of poly(adenosine diphosphate-ribose) polymerase (PARP), an end product of caspase activation, were determined by immunoblotting. Bcl-2 and Bax heterodimerization was examined by coimmunoprecipitation. All statistical tests were two-sided. RESULTS: Sensitivity to Fas-mediated apoptosis decreased with the tumorigenic potential of cells: MCF-10A cells were extremely susceptible, MCF-10AT cells were less susceptible, and MCF-10ATG3B cells were resistant. The percentage of apoptotic cells declined, from 24% to 8% to 6%, respectively. All lines produced Fas ligand (FasL) and had comparable levels of Fas receptor, FasL, Fas-associated death-domain protein, and caspases 3 and 6. Levels of caspase 8 were similar in MCF-10A and MCF-10AT cells but about 30% lower in MCF-10ATG3B cells (P>.01 but <.05). Levels of caspase 10 were about 20% lower in MCF-10AT cells (P>.005 but <.01) and about 59% lower in MCF-10ATG3B cells than in MCF-10A cells (P>.01 but <.05). PARP cleavage was detected in MCF-10A and MCF-10AT cells but not in MCF-10ATG3B cells. Levels of Bax, Bid, and Bak proteins were similar in all lines, but levels of Bcl-2 were lower in MCF-10AT and MCF-10ATG3B cells than in MCF-A cells, and Bcl-2-Bax heterodimerization progressively declined in the series. CONCLUSION: Resistance to Fas-mediated apoptosis appears to develop progressively in the MCF-10AT cell series.
BACKGROUND: Proliferative breast disease (PBD) may increase a woman's risk of developing breast cancer, perhaps by decreasing cellular sensitivity to apoptosis. To determine whether resistance to apoptosis develops during PBD, we investigated apoptosis initiated through the Fas pathway in a series of cell lines that recapitulates the morphologic changes of PBD in nude/beige mice. METHODS: The series of cell lines used was MCF-10A cells (parental preneoplastic human breast epithelial cells), MCF-10AT cells (transformed with T(24) Ha-ras), and MCF-10ATG3B cells (derivative cells that progress to carcinoma). Fas-mediated apoptosis, induced when a Fas monoclonal antibody bound to and activated the Fas receptor on these cells, was assessed morphologically and by flow cytometry. Levels of proteins involved in Fas-mediated apoptosis and cleavage of poly(adenosine diphosphate-ribose) polymerase (PARP), an end product of caspase activation, were determined by immunoblotting. Bcl-2 and Bax heterodimerization was examined by coimmunoprecipitation. All statistical tests were two-sided. RESULTS: Sensitivity to Fas-mediated apoptosis decreased with the tumorigenic potential of cells: MCF-10A cells were extremely susceptible, MCF-10AT cells were less susceptible, and MCF-10ATG3B cells were resistant. The percentage of apoptotic cells declined, from 24% to 8% to 6%, respectively. All lines produced Fas ligand (FasL) and had comparable levels of Fas receptor, FasL, Fas-associated death-domain protein, and caspases 3 and 6. Levels of caspase 8 were similar in MCF-10A and MCF-10AT cells but about 30% lower in MCF-10ATG3B cells (P>.01 but <.05). Levels of caspase 10 were about 20% lower in MCF-10AT cells (P>.005 but <.01) and about 59% lower in MCF-10ATG3B cells than in MCF-10A cells (P>.01 but <.05). PARP cleavage was detected in MCF-10A and MCF-10AT cells but not in MCF-10ATG3B cells. Levels of Bax, Bid, and Bak proteins were similar in all lines, but levels of Bcl-2 were lower in MCF-10AT and MCF-10ATG3B cells than in MCF-A cells, and Bcl-2-Bax heterodimerization progressively declined in the series. CONCLUSION: Resistance to Fas-mediated apoptosis appears to develop progressively in the MCF-10AT cell series.
Authors: M Murphy; M J E M F Mabruk; P Lenane; A Liew; P McCann; A Buckley; C O Flatharta; D Hevey; P Billet; W Robertson; S Javed; M Leader; E Kay; G M Murphy Journal: J Clin Pathol Date: 2002-11 Impact factor: 3.411
Authors: Srinivas Malladi; Kishore V L Parsa; Deepthi Bhupathi; María A Rodríguez-González; Juan A Conde; Pallavi Anumula; Hannah E Romo; Cheryl J Claunch; Rafael P Ballestero; Maribel González-García Journal: Mol Cell Biochem Date: 2011-01-21 Impact factor: 3.396
Authors: Mia M Gaudet; Roger L Milne; Angela Cox; Nicola J Camp; Ellen L Goode; Manjeet K Humphreys; Alison M Dunning; Jonathan Morrison; Graham G Giles; Gianluca Severi; Laura Baglietto; Dallas R English; Fergus J Couch; Janet E Olson; Xianshu Wang; Jenny Chang-Claude; Dieter Flesch-Janys; Sascha Abbas; Ramona Salazar; Arto Mannermaa; Vesa Kataja; Veli-Matti Kosma; Annika Lindblom; Sara Margolin; Tuomas Heikkinen; Kati Kämpjärvi; Kirsimari Aaltonen; Heli Nevanlinna; Natalia Bogdanova; Irina Coinac; Peter Schürmann; Thilo Dörk; Claus R Bartram; Rita K Schmutzler; Sandrine Tchatchou; Barbara Burwinkel; Hiltrud Brauch; Diana Torres; Ute Hamann; Christina Justenhoven; Gloria Ribas; José I Arias; Javier Benitez; Stig E Bojesen; Børge G Nordestgaard; Henrik L Flyger; Julian Peto; Olivia Fletcher; Nichola Johnson; Isabel Dos Santos Silva; Peter A Fasching; Matthias W Beckmann; Reiner Strick; Arif B Ekici; Annegien Broeks; Marjanka K Schmidt; Flora E van Leeuwen; Laura J Van't Veer; Melissa C Southey; John L Hopper; Carmel Apicella; Christopher A Haiman; Brian E Henderson; Loic Le Marchand; Laurence N Kolonel; Vessela Kristensen; Grethe Grenaker Alnaes; David J Hunter; Peter Kraft; David G Cox; Susan E Hankinson; Caroline Seynaeve; Maaike P G Vreeswijk; Rob A E M Tollenaar; Peter Devilee; Stephen Chanock; Jolanta Lissowska; Louise Brinton; Beata Peplonska; Kamila Czene; Per Hall; Yuqing Li; Jianjun Liu; Sabapathy Balasubramanian; Saeed Rafii; Malcolm W R Reed; Karen A Pooley; Don Conroy; Caroline Baynes; Daehee Kang; Keun-Young Yoo; Dong-Young Noh; Sei-Hyun Ahn; Chen-Yang Shen; Hui-Chun Wang; Jyh-Cherng Yu; Pei-Ei Wu; Hoda Anton-Culver; Argyrios Ziogoas; Kathleen Egan; Polly Newcomb; Linda Titus-Ernstoff; Amy Trentham Dietz; Alice J Sigurdson; Bruce H Alexander; Parveen Bhatti; Kristina Allen-Brady; Lisa A Cannon-Albright; Jathine Wong; Georgia Chenevix-Trench; Amanda B Spurdle; Jonathan Beesley; Paul D P Pharoah; Doug F Easton; Montserrat Garcia-Closas Journal: Cancer Epidemiol Biomarkers Prev Date: 2009-05 Impact factor: 4.254