Intracrine PTHrP protects against serum starvation-induced apoptosis and regulates the cell cycle in MCF-7 breast cancer cells.
PTHrP is secreted by breast cancer cells in vivo and in vitro. In the breast cancer cell line MCF-7, PTHrP overexpression is associated with increased mitogenesis. We used this cell line to study the mechanism for the proliferative effects of PTHrP. Clonal MCF-7 lines were established overexpressing wild-type PTHrP or PTHrP mutated in the nuclear localization signal (NLS). Mutation of the NLS negated the proliferative effects and nuclear trafficking of PTHrP, indicating that increased mitogenesis is mediated via an intracrine pathway. Cells overexpressing wild-type PTHrP were enriched in G2 + M stage of the cell cycle compared with cells overexpressing NLS-mutated PTHrP, indicating an intracrine role for PTHrP in cell cycle regulation. Wild-type PTHrP also protected MCF-7 cells from serum starvation-induced apoptosis. Cells overexpressing wild-type PTHrP showed significantly greater cell survival than cells overexpressing NLS-mutated PTHrP. The ratios of the apoptosis-regulating proteins Bcl-2 to Bax and Bcl-x(L) to Bax were higher in cells overexpressing wild-type, but not NLS-mutated, PTHrP compared with control cells. These findings suggest that the proliferative effects of PTHrP in breast cancer cells are mediated through regulation of the cell cycle and apoptosis, and that controlling PTHrP production in breast cancer may be therapeutically useful.
Tovar Sepulveda
Veronica A
VA
Department of Pharmacology and Toxicology and Sealy Center for Molecular Science, University of Texas Medical Branch, Galveston, Texas 77555, USA.
Shen
Xiaoli
X
Falzon
Miriam
M
D017209:Apoptosis / Q000187:drug effects*; D015152:Blotting, Northern; D001943:Breast Neoplasms / Q000473:pathology*; D002452:Cell Count; D002453:Cell Cycle / Q000187:drug effects; D002467:Cell Nucleus / Q000502:physiology / Q000648:ultrastructure; D016895:Culture Media, Serum-Free; D005260:Female; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D019254:Genes, bcl-2 / Q000235:genetics; D006801:Humans; D009154:Mutation / Q000235:genetics; D044162:Parathyroid Hormone-Related Protein; D010957:Plasmids / Q000235:genetics; D011506:Proteins / Q000235:genetics / Q000502:physiology*; D012333:RNA, Messenger / Q000096:biosynthesis; D015398:Signal Transduction / Q000502:physiology; D013936:Thymidine / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured
D016895:Culture Media, Serum-Free; C470121:PTHLH protein, human; D044162:Parathyroid Hormone-Related Protein; D011506:Proteins; D012333:RNA, Messenger; D013936:Thymidine