Preprint / Version 1

Withaferin a Triggers Apoptosis and DNA Damage in Bladder Cancer J82 Cells through Oxidative Stress

Authors

  • Tsu-Ming Chien Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; [email protected]
  • Wen-Jeng Wu Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; [email protected]
  • Tzu-Jung Yu Graduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; [email protected] (C.-H.Y.); [email protected] (T.-J.Y.)
  • Chia-Hung Yen Graduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; [email protected] (C.-H.Y.); [email protected] (T.-J.Y.)
  • Bi-Wen Yeh Department of Urology, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; [email protected]
  • Hui-Ru Wang Department of Biomedical Science and Environmental Biology, College of Life Science, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; [email protected] (Y.-T.C.); [email protected] (Y.-C.Y.); [email protected] (H.-R.W.)
  • Yun-Chiao Yeh Department of Biomedical Science and Environmental Biology, College of Life Science, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; [email protected] (Y.-T.C.); [email protected] (Y.-C.Y.); [email protected] (H.-R.W.)
  • Ya-Ting Chuang Department of Biomedical Science and Environmental Biology, College of Life Science, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; [email protected] (Y.-T.C.); [email protected] (Y.-C.Y.); [email protected] (H.-R.W.)
  • Kuang-Han Wu Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; [email protected]
  • Hsueh-Wei Chang Department of Biomedical Science and Environmental Biology, College of Life Science, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; [email protected] (Y.-T.C.); [email protected] (Y.-C.Y.); [email protected] (H.-R.W.)

Keywords:

Withaferin A, bladder cancer, DNA damage, apoptosis, oxidative stress

Abstract

Withaferin A (WFA), the Indian ginseng bioactive compound, exhibits an antiproliferation effect on several kinds of cancer, but it was rarely reported in bladder cancer cells. This study aims to assess the anticancer effect and mechanism of WFA in bladder cancer cells. WFA shows antiproliferation to bladder cancer J82 cells based on the finding of the MTS assay. WFA disturbs cell cycle progression associated with subG1 accumulation in J82 cells. Furthermore, WFA triggers apoptosis as determined by flow cytometry assays using annexin V/7-aminoactinomycin D and pancaspase detection. Western blotting also supports WFA-induced apoptosis by increasing cleavage of caspases 3, 8, and 9 and poly ADP-ribose polymerase. Mechanistically, WFA triggers oxidative stress-association changes, such as the generation of reactive oxygen species and mitochondrial superoxide and diminishment of the mitochondrial membrane potential, in J82 cells. In response to oxidative stresses, mRNA for antioxidant signaling, such as nuclear factor erythroid 2-like 2 (NFE2L2), catalase (CAT), superoxide dismutase 1 (SOD1), thioredoxin (TXN), glutathione-disulfide reductase (GSR), quinone dehydrogenase 1 (NQO1), and heme oxygenase 1 (HMOX1), are overexpressed in J82 cells. In addition, WFA causes DNA strand breaks and oxidative DNA damages. Moreover, the ROS scavenger N-acetylcysteine reverts all tested WFA-modulating effects. In conclusion, WFA possesses anti-bladder cancer effects by inducing antiproliferation, apoptosis, and DNA damage in an oxidative stress-dependent manner. Keywords: Withaferin A, bladder cancer, DNA damage, apoptosis, oxidative stress

Author Biographies

Tsu-Ming Chien, Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; [email protected]

Department of Urology, Kaohsiung Medical University Hospital, Kaohsiung 80708, Taiwan

Wen-Jeng Wu, Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; [email protected]

Department of Urology, Kaohsiung Medical University Hospital, Kaohsiung 80708, Taiwan

Bi-Wen Yeh, Department of Urology, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; [email protected]

Department of Urology, Kaohsiung Medical University Hospital, Kaohsiung 80708, Taiwan

Hsueh-Wei Chang, Department of Biomedical Science and Environmental Biology, College of Life Science, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; [email protected] (Y.-T.C.); [email protected] (Y.-C.Y.); [email protected] (H.-R.W.)

Center for Cancer Research, Kaohsiung Medical University, Kaohsiung 80708, Taiwan

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