Natural Antioxidant Resveratrol Suppresses Uterine Fibroid Cell Growth and Extracellular Matrix Formation In Vitro and In Vivo

Hsin-Yuan Chen; Po-Han Lin; Yin-Hwa Shih; Kei-Lee Wang; Yong-Han Hong; Tzong-Ming Shieh; Tsui-Chin Huang; Shih-Min Hsia

doi:10.3390/antiox8040099

Natural Antioxidant Resveratrol Suppresses Uterine Fibroid Cell Growth and Extracellular Matrix Formation In Vitro and In Vivo

Antioxidants (Basel). 2019 Apr 12;8(4):99. doi: 10.3390/antiox8040099.

Authors

Hsin-Yuan Chen¹, Po-Han Lin², Yin-Hwa Shih³, Kei-Lee Wang⁴, Yong-Han Hong⁵, Tzong-Ming Shieh⁶, Tsui-Chin Huang⁷, Shih-Min Hsia^{8

9

10

11}

Affiliations

¹ School of Nutrition and Health Sciences, College of Nutrition, Taipei Medical University, Taipei 11031, Taiwan. hsin246@gmail.com.
² School of Nutrition and Health Sciences, College of Nutrition, Taipei Medical University, Taipei 11031, Taiwan. phlin@tmu.edu.tw.
³ Department of Healthcare Administration, Asia University, Taichung 41354, Taiwan. evashih@asia.edu.tw.
⁴ Department of Nursing, Ching Kuo Institute of Managemnet and Health, Keelung 20301, Taiwan. kellywang@tmu.edu.tw.
⁵ Department of Nutrition, I-Shou University, Kaohsiung 84001, Taiwan. yonghan@isu.edu.tw.
⁶ Department of Dental Hygiene, College of Health Care, China Medical University, Taichung 40402, Taiwan. tmshieh@mail.cmu.edu.tw.
⁷ PhD Program for Cancer Biology and Drug Discovery, College of Medical Science and Technology, Taipei Medical University and Academia Sinica, Taipei 11031, Taiwan. tsuichin@tmu.edu.tw.
⁸ School of Nutrition and Health Sciences, College of Nutrition, Taipei Medical University, Taipei 11031, Taiwan. bryanhsia@tmu.edu.tw.
⁹ Graduate Institute of Metabolism and Obesity Sciences, College of Nutrition, Taipei Medical University, Taipei 11031, Taiwan. bryanhsia@tmu.edu.tw.
¹⁰ School of Food and Safety, Taipei Medical University, Taipei 11031, Taiwan. bryanhsia@tmu.edu.tw.
¹¹ Nutrition Research Center, Taipei Medical University Hospital, Taipei 11031, Taiwan. bryanhsia@tmu.edu.tw.

Abstract

Resveratrol (RSV) is a polyphenolic phytoalexin found in peanuts, grapes, and other plants. Uterine fibroids (UF) are benign growths that are enriched in extracellular matrix (ECM) proteins. In this study, we aimed to investigate the effects of RSV on UF using in vivo and in vitro approaches. In mouse xenograft models, tumors were implanted through the subcutaneous injection of Eker rat-derived uterine leiomyoma cells transfected with luciferase (ELT-3-LUC) in five-week-old female nude (Foxn1^nu) mice. When the tumors reached a size of 50-100 mm³, the mice were randomly assigned to intraperitoneal treatment with RSV (10 mg·kg^-1) or vehicle control (dimethyl sulfoxide). Tumor tissues were assayed using an immunohistochemistry analysis. We also used primary human leiomyoma cells as in vitro models. Cell viability was determined using the sodium bicarbonate and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The protein expression was assayed using Western blot analysis. The messenger ribonucleic acid (mRNA) expression was assayed using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Cell apoptosis was assayed using Annexin V-fluorescein isothiocyanate (FITC) and propidium iodide (PI) and Hoechst 33342 staining. RSV significantly suppressed tumor growth in vivo and decreased the proportion of cells showing expression of proliferating cell nuclear antigen (PCNA) and α-smooth muscle actin (α-SMA). In addition, RSV decreased the protein expression of PCNA, fibronectin, and upregulated the ratio of Bax (Bcl-2-associated X) and Bcl-2 (B-cell lymphoma/leukemia 2) in vivo. Furthermore, RSV reduced leiomyoma cell viability, and decreased the mRNA levels of fibronectin and the protein expression of collagen type 1 (COL1A1) and α-SMA (ECM protein marker), as well as reducing the levels of β-catenin protein. RSV induced apoptosis and cell cycle arrest at sub-G1 phase. Our findings indicated the inhibitory effects of RSV on the ELT-3-LUC xenograft model and indicated that RSV reduced ECM-related protein expression in primary human leiomyoma cells, demonstrating its potential as an anti-fibrotic therapy for UF.

Keywords: ELT-3-LUC xenograft model; extracellular matrix; resveratrol; uterine fibroids.