Export
Role of calpain-9 and PKC-delta in the apoptotic mechanism of lumen formation in CEACAM1 transfected breast epithelial cells
Periodical: Exp Cell Res ISBN: 1090-2422 (Electronic)
Date: 2009/11/17
Language: Eng
Authors:Chen, C. J., Nguyen, T., Shively, J. E.
A copy of this paper may be available for free: Google Scholar Search 
Abstract
CEACAM1-4S (carcinoembryonic antigen-related cell adhesion molecule 1) is a type I membrane protein with a short (12 amino acid) cytoplasmic tail. Wild type CEACAM1-4S transfected MCF7 cells form glands with lumena when grown in 3D culture, while null mutations of two putative phosphorylation sites (T457A and S459A) in the cytoplasmic domain fail to undergo lumen formation. When gene chip analysis was performed on mRNA isolated from both wild type and T457A,S459A mutated CEACAM1-4S transfected MCF7 cells grown in 3D culture, calpain-9 (CAPN9) was identified out of over 400 genes with a >2 log 2 difference as a potential inducer of lumen formation. Inhibition of CAPN9 expression in MCF7/CEACAM1-4S cells by RNAi or by calpeptin or PD150606 inhibited lumen formation. Transfection of CAPN9 into wild type MCF7 cells restores lumen formation demonstrating that calpain-9 may plays a critical role in lumen formation. Additionally, we demonstrate that the apoptosis related kinase, PKC-delta is activated by proteolytic cleavage during lumen formation exclusively in wild type CEACAM1-4S transfected MCF7 cells grown in 3D culture, and that lumen formation is inhibited by either RNAi to PKC-delta or by the PKC-delta inhibitor rottlerin.
CEACAM1-4S (carcinoembryonic antigen-related cell adhesion molecule 1) is a type I membrane protein with a short (12 amino acid) cytoplasmic tail. Wild type CEACAM1-4S transfected MCF7 cells form glands with lumena when grown in 3D culture, while null mutations of two putative phosphorylation sites (T457A and S459A) in the cytoplasmic domain fail to undergo lumen formation. When gene chip analysis was performed on mRNA isolated from both wild type and T457A,S459A mutated CEACAM1-4S transfected MCF7 cells grown in 3D culture, calpain-9 (CAPN9) was identified out of over 400 genes with a >2 log 2 difference as a potential inducer of lumen formation. Inhibition of CAPN9 expression in MCF7/CEACAM1-4S cells by RNAi or by calpeptin or PD150606 inhibited lumen formation. Transfection of CAPN9 into wild type MCF7 cells restores lumen formation demonstrating that calpain-9 may plays a critical role in lumen formation. Additionally, we demonstrate that the apoptosis related kinase, PKC-delta is activated by proteolytic cleavage during lumen formation exclusively in wild type CEACAM1-4S transfected MCF7 cells grown in 3D culture, and that lumen formation is inhibited by either RNAi to PKC-delta or by the PKC-delta inhibitor rottlerin.
Keywords
Animals, Biocompatible Materials/ chemistry, Cell Culture Techniques/methods, Cells, Cultured, Crystallization/methods, Extracellular Matrix Proteins/ metabolism/ ultrastructure, Extracellular Matrix/chemistry, Humans, Materials Testing, Mesenchymal Stem Cells/ ultrastructure, Mice, Molecular Conformation, Particle Size, Polyglycolic Acid/ chemistry, Surface Properties, Synchrotrons, Tissue Engineering/ methods, Tomography, X-Ray Computed
Animals, Biocompatible Materials/ chemistry, Cell Culture Techniques/methods, Cells, Cultured, Crystallization/methods, Extracellular Matrix Proteins/ metabolism/ ultrastructure, Extracellular Matrix/chemistry, Humans, Materials Testing, Mesenchymal Stem Cells/ ultrastructure, Mice, Molecular Conformation, Particle Size, Polyglycolic Acid/ chemistry, Surface Properties, Synchrotrons, Tissue Engineering/ methods, Tomography, X-Ray Computed
3DCellculture.com's MAMI
search attributes
CellLine: MCF-7
Morphology: Cancerous
Origin: Breast
Species: Human
Scaffold Form: gel/hydrogelMorphology: Cancerous
Origin: Breast
Species: Human
Scaffold Material: Matrigel (TM)