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Connective tissue growth factor induces apoptosis in human breast cancer cell line MCF-7
K Hishikawa1, B S Oemar, F C Tanner
1Department of Pharmacology, Teikyo University School of Medicine, Tokyo 173-8605, Japan. hisikawa@med.teikyo-u.ac.jp
Abstract:
Connective tissue growth factor (CTGF) is a member of an emerging CCN gene family that is implicated in various diseases associated with fibro-proliferative disorder including scleroderma and atherosclerosis. The function of CTGF in human cancer is largely unknown. We now show that CTGF induces apoptosis in the human breast cancer cell line MCF-7. CTGF mRNA was completely absent in MCF-7 but strongly induced by treatment with transforming growth factor beta (TGF-beta). TGF-beta by itself induced apoptosis in MCF-7, and this effect was reversed by co-treatment with CTGF antisense oligonucleotide. Overexpression of CTGF gene in transiently transfected MCF-7 cells significantly augmented apoptosis. Moreover, recombinant CTGF protein significantly enhanced apoptosis in MCF-7 cells as evaluated by DNA fragmentation, Tdt-mediated dUTP biotin nick end-labeling staining, flow cytometry analysis, and nuclear staining using Hoechst 33258. Finally, recombinant CTGF showed no effect on Bax protein expression but significantly reduced Bcl2 protein expression. Taken together, these results suggest that CTGF is a major inducer of apoptosis in the human breast cancer cell line MCF-7 and that TGF-beta-induced apoptosis in MCF-7 cells is mediated, in part, by CTGF.
Insights
Connective tissue growth factor (CTGF) induces apoptosis in human breast cancer cells (MCF-7). Transforming growth factor beta (TGF-beta) also triggers apoptosis, partly mediated by CTGF.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Connective tissue growth factor (CTGF) is part of the CCN gene family, linked to fibro-proliferative disorders.
- The role of CTGF in human cancer, particularly breast cancer, remains largely uncharacterized.
- Understanding CTGF's function is crucial for developing novel cancer therapies.
Purpose of the Study:
- To investigate the role and function of CTGF in human breast cancer.
- To determine if CTGF influences apoptosis in the MCF-7 breast cancer cell line.
- To elucidate the relationship between CTGF, transforming growth factor beta (TGF-beta), and apoptosis in MCF-7 cells.
Main Methods:
- Assessed CTGF mRNA expression in MCF-7 cells.
- Investigated the effect of TGF-beta on CTGF expression and apoptosis.
- Utilized CTGF antisense oligonucleotides to block CTGF function.
- Examined apoptosis induction via recombinant CTGF protein using DNA fragmentation, TUNEL staining, flow cytometry, and nuclear staining.
- Analyzed Bax and Bcl2 protein expression levels.
Main Results:
- CTGF mRNA was absent in MCF-7 cells but induced by TGF-beta.
- TGF-beta treatment induced apoptosis in MCF-7 cells, an effect reversed by CTGF antisense oligonucleotide.
- Overexpression of CTGF and treatment with recombinant CTGF protein significantly enhanced apoptosis in MCF-7 cells.
- Recombinant CTGF reduced Bcl2 protein expression but did not affect Bax expression.
Conclusions:
- CTGF is a significant inducer of apoptosis in the human breast cancer cell line MCF-7.
- TGF-beta-induced apoptosis in MCF-7 cells is, in part, mediated by CTGF.
- CTGF may represent a therapeutic target for breast cancer treatment.