H4K20me3 is increased in human bladder cancer tissues and is upregulated by reactive oxygen species in bladder cancer cell lines

Sotida  Kayem; Suchittra  Phoyen; Patcharawalai  Whongsiri; Chaowat  Pimratana; Udomsak  Wijitsettakul; Chanchai  Boonla

Authors

Sotida Kayem Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand
Suchittra Phoyen Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand
Patcharawalai Whongsiri Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand and Center of Excellence in DNA Barcoding of Thai Medicinal Plants, Chulalongkorn University, Bangkok, Thailand
Chaowat Pimratana Division of Urology, Buriram Hospital, Buriram, Thailand
Udomsak Wijitsettakul Division of Urology, Buriram Hospital, Buriram, Thailand
Chanchai Boonla Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand

Keywords:

Bladder cancer, H4K20me3, histone methylation, oxidative stress, ROS

Abstract

Background: Previously, we demonstrated that reactive oxygen species (ROS) triggered oxidative stress,enhanced tumor aggressiveness, and induced a profound change in histone methylation in bladder cancer (BlCa) and reported H4K20me3 upregulation by ROS in hepatocellular carcinoma.

Objective: This study aimed to investigate whether H4K20me3 was upregulated in human BlCa tissues and determine if ROS could induce H4K20me3 formation in BlCa cell lines.

Methods: Immunohistochemical (IHC) staining was performed in 37 BlCa sections and 6 adjacent noncancerous tissues (controls). ROS upregulated H4K20me3 was investigated in three BlCa cell lines: UM-UC-3, VM-CUB- 1, and TCCSUP.

Results: The H4K20me3 levels in BlCa tissues increased relative to that in adjacent noncancerous tissues.The IHC score of the H4K20me3 level in BlCa tissues was significantly higher than that in noncancerous controls. H2O2(ROS representative) at 50, 100, and 200 mM significantly induced oxidative stress in UM-UC-3, VM-CUB-1, and TCCSUP cells, respectively, but did not significantly alter cell survival. Western blot and immunofluorescent staining results showed that H2O2treatment markedly increased H4K20me3 formation in all three cell lines.

Conclusion: This study demonstrated that the H4K20me3 levels in BlCa tissues obtained from Thai patients with BlCa increased compared with the levels in adjacent noncancerous tissues. Evidently, ROS upregulated H4K20me3 formation in BlCa cell lines. Perhaps, ROS induced the expression of histone methyltransferases that further caused an increase in H4K20me3 formation. The induction of tumor progression by ROS is well recognized; however, whether ROS induced BlCa progression is mediated by H4K20me3 formation remains to be elucidated.

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