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Inhibition of peptidyl–prolyl cis/trans isomerase Pin1 induces cell cycle arrest and apoptosis in vascular smooth muscle cells

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Abstract

This study was undertaken to determine the in vitro effect of lentivirus-mediated siPin1 on cell cycle and apoptosis of vascular smooth muscle cells (VSMCs). Further we sought to provide insight into the mechanisms behind these processes. Human umbilical artery smooth muscle cells (HUASMCs) were transfected with lentiviral siPin1. Real-time RT–PCR and Western blotting were used to examine Pin1 mRNA and protein expression. MTT and [3H]thymidine incorporation assays were employed to observe cell proliferation status. The apoptotic rate and cell cycle were analyzed by Hoechst33258 staining and flow cytometry. Finally we measured the expression of cyclin D1, β-catenin, CDK4, cytochrome c, procaspase-3, cleaved caspase-3, procaspase-9, cleaved caspase-9, Bcl-2, Bax, STAT3, phosphorylated STAT3 and VEGF in lentiviral siPin1 infected VSMCs. Lentivirus-mediated siPin1 effectively diminished endogenous Pin1 expression in VSMCs resulting in cell cycle arrest and enhancement of apoptosis. This was accompanied by downregulation of cyclin D1, β-catenin, CDK4, increase of Bax/Bcl-2 ratio, release of cytochrome c, and activation of caspase-3 and -9. We concluded that this effect was mediated, at least in part, via the β-catenin/cyclin D1/CDK4 cascade, and that the mitochondrial pathway was responsible for VSMC apoptosis in the absence of Pin1. Our observations raised the possibility that Pin1 might be a potential therapeutic target to prevent stenosis.

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Acknowledgments

This work was supported by grants from the research program of Shanghai Renji Hospital. Special thanks go to the staff obstetricians of the department of obstetrics and gynecology at Shanghai Renji Hospital for providing the umbilical cords. We wish to thank Dennis Cho, Jay Wang and Susana Huang (The Schulich School of Medicine & Dentistry, University of Western Ontario) for their assistance in proofreading this manuscript.

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  1. Department of Vascular Surgery, Renji Hospital, Shanghai Jiaotong University, College of Medicine, 145 Hao, Shandong Zhong Road, Shanghai, China

    Lei Lv, Zhaoxiong Zhou, Xiaozhong Huang, Yingping Zhao, Lan Zhang, Yaxue Shi, Mingli Sun & Jiwei Zhang

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  1. Lei Lv
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  2. Zhaoxiong Zhou
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  3. Xiaozhong Huang
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  8. Jiwei Zhang
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Correspondence to Zhaoxiong Zhou or Jiwei Zhang.

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Lv, L., Zhou, Z., Huang, X. et al. Inhibition of peptidyl–prolyl cis/trans isomerase Pin1 induces cell cycle arrest and apoptosis in vascular smooth muscle cells. Apoptosis 15, 41–54 (2010). https://doi.org/10.1007/s10495-009-0409-8

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