The loss-of-function mutations and down-regulated expression of ASB3 gene promote the growth and metastasis of colorectal cancer cells

Abstract

BackgroundAnkyrin repeat and SOCS box protein 3 (ASB3) is a member of ASB family and contains ankyrin repeat sequence and SOCS box domain. Previous studies indicated that it mediates the ubiquitination and degradation of tumor necrosis factor receptor 2 and is likely involved in inflammatory responses. However, its effects on oncogenesis are unclear. This study aimed to investigate the effects of ASB3 on the growth and metastasis of colorectal cancer (CRC).MethodsWe used next-generation sequencing or Sanger sequencing to detect ASB3 mutations in CRC specimens or cell lines, and used real-time quantitative polymerase chain reaction, Western blotting, and immunohistochemical or immunofluorescence assay to determine gene expression. We evaluated cell proliferation by MTT and colony formation assays, tested cell cycle distribution by flow cytometry, and assessed cell migration and invasion by transwell and wound healing assays. We also performed nude mouse experiments to evaluate tumorigenicity and hepatic metastasis potential of tumor cells.ResultsWe found that ASB3 gene was frequently mutated (5.3%) and down-regulated (70.4%) in CRC cases. Knockdown of endogenous ASB3 expression promoted CRC cell proliferation, migration, and invasion in vitro and facilitated tumorigenicity and hepatic metastasis in vivo. Conversely, the ectopic overexpression of wild-type ASB3, but not that of ASB3 mutants that occurred in clinical CRC tissues, inhibited tumor growth and metastasis. Further analysis showed that ASB3 inhibited CRC metastasis likely by retarding epithelial-mesenchymal transition, which was characterized by the up-regulation of β-catenin and E-cadherin and the down-regulation of transcription factor 8, N-cadherin, and vimentin.Conclusion ASB3 dysfunction resulted from gene mutations or down-regulated expression frequently exists in CRC and likely plays a key role in the pathogenesis and progression of CRC.

Anxiety Associated Increased CpG Methylation in the Promoter of Asb1: A Translational Approach Evidenced by Epidemiological and Clinical Studies and a Murine Model

“…Polymorphisms in this member of the IL-1 superfamily were shown to be predictive of depression and anxiety through altered amygdala reactivity of threat perception in women but not in men ( Swartz et al , 2017 ). While the role of ASB family members is not well described ( Chung et al , 2005 ), one family member ASB3 has been shown to regulate responses to the pro-inflammatory cytokine tumor necrosis factor (TNF)-alpha by mediating the degradation of its receptor TNF-R2 ( Chung et al , 2005 ) and its gene expression is downregulated in colorectal cancer ( Du et al , 2017 ). The impact of stress-induced Asb1 expression on peripheral inflammation was further supported in our mouse model, where we observed strong correlation of Asb1 expression with expression of the IL-1B gene.…”

Up‐regulation of indoleamine 2,3‐dioxygenase 1 (IDO1) expression and catalytic activity is associated with immunosuppression and poor prognosis in penile squamous cell carcinoma patients

“…Western blotting was performed as previously described [24, 25]. Briefly, the cells mentioned above were collected and lysed in lysis buffer [24] and then concentrated by a BCA protein assay kit (Pierce Biotechnology, Rockford, IL, USA).…”

Molecular characterization and integrative genomic analysis of a panel of newly established penile cancer cell lines

“…The migration ability of cell lines was assessed using a monolayer wound healing assay 45 . Briefly, cells were seeded in 6-well plates and grown to confluence overnight.…”