Elucidation of the therapeutic role of nuclear respiratory factor-1 in the regulation of prostate cancer
闡明核呼吸因子-1在前列腺癌調節中的治療
Chun-Hsien Wu1,2,*, Chung-Hsien Chen1,2,*, Pei-fang Hsieh3,4, Chia-Lung Tsai1,2,, Shu-Fen Liu5, Tsung-Jen Hung6, Tzu-sui Hung7, Ching-Yu Hung1,2, Richard Chen-Yu Wu1,2 , Victor C. Lin1,2,#
1Department of Urology, E-Da Hospital, Kaohsiung, Taiwan
2School of Medicine for International Students, I-Shou University, Kaohsiung, Taiwan
3Graduate Institute of Biomedical Science, National Sun Yat-sen University, Kaohsiung, Taiwan.
4Graduate Institute of Medical Laboratory Science and Biotechnology, Chung Hwa University of Medical Technology, Tainan, Taiwan.
5Department of Internal Medicine, Kaohsiung Medical University Chung-Ho Memorial Hospital, Kaohsiung, Taiwan.
6Graduate Institute of Biomedical Science, Chung Hwa University of Medical Technology, Tainan, Taiwan.
7Department of sport, Health and Leisure, Chung Hwa University of Medical Technology, Taiwan
* Chun-Hsien Wu and Chung-Hsien Chen contributed equally to this work.
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Abstract
Introduction: The pathogenesis of prostate cancer cell might be associated with the dysregulation of cellular energy. Nuclear respiratory factor1 (NRF-1) functions as a transcription factor that activates the expression of key metabolic genes regulating cellular growth and nuclear genes required for mitochondrial respiration, and mitochondrial DNA transcription and replication. The prostate cancer cell regulatory role of NRF-1 will be extensively studied in this study.
Methods: We cultured human prostate cancer cell line (PC3) cells and overexpressed NRF-1(pcDNA-NRF-1) by the vector transfection to explore the effect on the cancer cell line.
Results: pcDNA-NRF-1 reduced the viability of prostate cancer cells by trypan blue exclusion assay. pcDNA-NRF-1 reduced the cell proliferation of prostate cancer cells by MTT assay. More importantly, pcDNA-NRF-1 induced cytotoxicity of prostate cancer cells by LDH assay. The TGF-β pathway is involved in prostate cancer progression. pcDNA-NRF-1 significantly reduced the expression of TGF-β1 secretion in PC3 cells. pcDNA-NRF-1 significantly decreased TGF-β1 signaling, in addition to inducing a marked increase in the down-regulation of Smad7 by western blot assay and immunofluorescence staining. pcDNA-NRF-1 decreased migration in prostate cancer cells. More importantly, pcDNA-NRF-1 decrease the expression of α-SMA (α-smooth muscle actin) and increase expression of E-cadherin by western blot and immunofluorescence.
Conclusions: We demonstrated that pcDNA-NRF-1 might act as a novel prostate cancer antagonist by down-regulating TGF-β1 signaling and modulating epithelial-mesanchymal transition (EMT). The study will be helpful for clinical urologists in treating prostate cancers.