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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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OTUB1 knockdown promotes apoptosis in melanoma cells by upregulating TRAIL expression

  • Lee, Bok-Soon (Department of Otolaryngology, School of Medicine, Ajou University) ;
  • Kang, Sung Un (Department of Otolaryngology, School of Medicine, Ajou University) ;
  • Huang, Mei (Department of Otolaryngology, School of Medicine, Ajou University) ;
  • Kim, Yeon Soo (Department of Otorhinolaryngology, College of Medicine, Konyang University Hospital, Konyang University Myunggok Medical Research Institute) ;
  • Lee, Young-Sun (School of Biosystem and Biomedical Science, College of Health Science, Korea University) ;
  • Park, Jae-Yong (School of Biosystem and Biomedical Science, College of Health Science, Korea University) ;
  • Kim, Chul-Ho (Department of Otolaryngology, School of Medicine, Ajou University)
  • Received : 2021.03.03
  • Accepted : 2021.08.02
  • Published : 2021.12.31
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Abstract

Melanoma, the most serious type of skin cancer, exhibits a high risk of metastasis. Although chemotherapeutic treatment for metastatic melanoma improves disease outcome and patient survival, some patients exhibit resistance or toxicity to the drug treatment regime. OTUB1 is a deubiquitinating enzyme overexpressed in several cancers. In this study, we investigated the effects of inhibiting OTUB1 expression on melanoma-cell proliferation and viability and identified the underlying molecular mechanism of action of OTUB1. We did endogenous OTUB1 knockdown in melanoma cells using short interfering RNA, and assessed the resulting phenotypes via MTT assays, Western blotting, and cell-cycle analysis. We identified differentially expressed genes between OTUB1-knockdown cells and control cells using RNA sequencing and confirmed them via Western blotting and reverse transcription polymerase chain reaction. Furthermore, we investigated the involvement of apoptotic and cell survival signaling pathways upon OTUB1 depletion. OTUB1 depletion in melanoma cells decreased cell viability and caused simultaneous accumulation of cells in the sub-G1 phase, indicating an increase in the apoptotic-cell population. RNA sequencing of OTUB1-knockdown cells revealed an increase in the levels of the apoptosis-inducing protein TRAIL. Additionally, OTUB1-knockdown cells exhibited increased sensitivity to PLX4032, a BRAF inhibitor, implying that OTUB1 and BRAF act collectively in regulating apoptosis. Taken together, our findings show that OTUB1 induces apoptosis of melanoma cells in vitro, likely by upregulating TRAIL, and suggest that approaches targeting OTUB1 can be developed to provide novel therapeutic strategies for treating melanoma.

Keywords

Acknowledgement

This research was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (Grant No: HR21C1003), and was supported by the National Research Foundation of Korea (Grant No: 2017R1D1A1B03028527 and 2018R1A2B3009008).

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