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CYT-Rx20 inhibits ovarian cancer cells in vitro and in vivo through oxidative stress-induced DNA damage and cell apoptosis

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Cancer Chemotherapy and Pharmacology Aims and scope Submit manuscript

Abstract

Purpose

The β-nitrostyrene family has been previously reported to possess anticancer property. However, the biological effects of β-nitrostyrenes on ovarian cancer and the underlying mechanisms involved remain unclear. In the present study, we synthesized a β-nitrostyrene derivative, CYT-Rx20 3′-hydroxy-4′-methoxy-β-methyl-β-nitrostyrene), and investigated its anticancer effects and the putative pathways of action in ovarian cancer.

Methods

The effects of CYT-Rx20 were analyzed using cell viability assay, reactive oxygen species (ROS) generation assay, FACS analysis, annexin V staining, immunostaining, comet assay, immunoblotting, soft agar assay, migration assay, nude mice xenograft study and immunohistochemistry.

Results

CYT-Rx20 induced cytotoxicity in ovarian cancer cells by promoting cell apoptosis via ROS generation and DNA damage. CYT-Rx20-induced cell apoptosis, ROS generation and DNA damage were reversed by thiol antioxidants. In addition, CYT-Rx20 inhibited ovarian cancer cell migration by regulating the expression of epithelial to mesenchymal transition (EMT) markers. In nude mice, CYT-Rx20 inhibited ovarian tumor growth accompanied by increased expression of DNA damage marker γH2AX and decreased expression of EMT marker Vimentin.

Conclusions

CYT-Rx20 inhibits ovarian cancer cells in vitro and in vivo, and has the potential to be further developed into an anti-ovarian cancer drug clinically.

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Acknowledgements

This study was supported by Grants from the Ministry of Health and Welfare (MOHW105-TDU-B-212-134007 and MOHW105-TDU-B-212-112016, Health and welfare surcharge of tobacco products) of Taiwan and National Health Research Institutes (NHRI-EX104-10212BI).

Author information

Authors and Affiliations

  1. Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan

    Yen-Yun Wang, Ya-Ling Hsu, Tsung-Chen Chi, Wan-Ling Huang & Shyng-Shiou F. Yuan

  2. Division of Oral Pathology and Maxillofacial Radiology, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan

    Yuk-Kwan Chen

  3. School of Dentistry, College of Dental Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

    Yuk-Kwan Chen

  4. Oral and Maxillofacial Imaging Center, College of Dental Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

    Yuk-Kwan Chen

  5. Department of Dermatology, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

    Stephen Chu-Sung Hu

  6. Department of Dermatology, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan

    Stephen Chu-Sung Hu

  7. Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

    Chun-Hao Tsai & Shyng-Shiou F. Yuan

  8. Graduate Institute of Natural Products, School of Traditional Chinese Medicine, Chang Gung University, Taoyuan, Taiwan

    Pei-Wen Hsieh

  9. Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan

    Pei-Wen Hsieh

  10. Department of Anesthesiology, Chang Gung Memorial Hospital, Taoyuan, Taiwan

    Pei-Wen Hsieh

  11. Translational Research Center, Kaohsiung Medical University Hospital, Kaohsiung Medical University, No.100, Tzyou 1st Road, 807, Kaohsiung, Taiwan

    Shyng-Shiou F. Yuan

  12. Department of Obstetrics and Gynecology, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan

    Shyng-Shiou F. Yuan

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  1. Yen-Yun Wang
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Corresponding author

Correspondence to Shyng-Shiou F. Yuan.

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Conflict of interest

Yen-Yun Wang, Yuk-Kwan Chen, Stephen Chu-Sung Hu, Ya-Ling Hsu, Chun-Hao Tsai, Tsung-Chen Chi, Wan-Ling Huang, Pei-Wen Hsieh, and Shyng-Shiou F. Yuan have declared no conflict of interest.

Ethical approval

The animal studies were approved by the Institutional Animal Care and Use Committee (IACUC No. 102009) of Kaohsiung Medical University, Taiwan. Animal experiments were approved by the Laboratory Animal Ethics Committee of Kaohsiung Medical University.

Electronic supplementary material

Below is the link to the electronic supplementary material.

280_2017_3330_MOESM1_ESM.pdf

Supplementary Fig. 1 Chemical structures of the β-nitrostyrene derivatives CYT-Rx20, CYT-Rx21, CYT-Rx22, CYT-Rx44, CYT-Rx45, CYT-Rx46, and CYT-Rx47 (PDF 43 kb)

280_2017_3330_MOESM2_ESM.pdf

Supplementary Fig. 2a Cells were treated with CYT-Rx20 for 1 h and ROS level was determined by staining with H2DCFDA fluorescent dye and analysis by flow cytometry. b Cells were treated with various concentrations of CYT-Rx20 (2 μg/ml) for 24 h prior to the determination of DNA damage by neutral comet assay. c Effect of CYT-Rx20 on induction of γ-H2AX focus formation in ovarian cancer cells. After treatment with the indicated concentrations of CYT-Rx20 (2 μg/ml) for 24 h, cells were fixed and incubated with antibodies against γ-H2AX, followed by secondary antibodies conjugated with the fluorochrome FITC. Nuclei were stained with DAPI. After immunostaining, cells were viewed by fluorescence microscope (original magnification × 1000) (PDF 218 kb)

280_2017_3330_MOESM3_ESM.pdf

Supplementary Fig. 3 MDAH 2774, PA-1, and SKOV3 cells were treated with various concentrations of CYT-Rx20 for 24 h and then cell viability was assessed by the XTT colorimetric assay (PDF 31 kb)

280_2017_3330_MOESM4_ESM.pdf

Supplementary Fig. 4 a The body weights of the nude mice included in this study were measured every week. b Hematoxylin and eosin staining of the tissues from mice organs. The representative photographs were shown with ×200 magnification. Bar represents 200 μm (PDF 466 kb)

Supplementary material 5 (PDF 28 kb)

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Wang, YY., Chen, YK., Hu, S.CS. et al. CYT-Rx20 inhibits ovarian cancer cells in vitro and in vivo through oxidative stress-induced DNA damage and cell apoptosis. Cancer Chemother Pharmacol 79, 1129–1140 (2017). https://doi.org/10.1007/s00280-017-3330-9

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  • DOI: https://doi.org/10.1007/s00280-017-3330-9

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