Ets1 and heat shock factor 1 regulate transcription of the Transformer 2β gene in human colon cancer cells
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Transformer (Tra) 2β is a member of the serine/arginine-rich (SR)-like protein family that regulates alternative splicing of numerous genes in a concentration-dependent manner. Several types of cancer cells up-regulate Tra2β expression, while the regulatory mechanism of Tra2β expression remains to be elucidated. In this study, we examined the transcriptional regulation and possible functions of Tra2β in human colon cancer cells.
We cloned 959 bp-upstream of the human TRA2β 5′-flank into luciferase constructs. Chromatin immunoprecipitation (ChIP) was employed to identify crucial cis element(s) and trans activator(s) of the TRA2β promoter. Tra2β expression in the human colon and colon cancer tissues was examined by immunohistochemistry.
In response to sodium arsenite, colon cancer cells (HCT116) increased levels of TRA2β1 mRNA encoding a functional, full-length Tra2β with a peak around 6 h without changing its mRNA stability. Transient expression assays using a reporter gene driven by serially truncated TRA2β promoters and Chip assay demonstrated that an Ets1-binding site present at −64 to −55 bp was crucial for basal transcription, while three heat shock elements (HSEs) located at −145 to −99 bp mediated the oxidant-induced transactivation of TRA2β. Tra2β knockdown caused apoptosis of HCT116 cells. Tra2β were preferentially expressed in proliferative compartment of normal human colonic glands and adenocarcinomas, where Ets1 and heat shock factor 1 were also highly expressed.
Our results suggest that oxidative stress-responsive Tra2β may play an important role in colon cancer growth.
KeywordsTransformer 2β Colon cancer Oxidative stress Ets1 Heat shock factor 1
This study was supported by Grants-in Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan (#22790649, 24659370 to Y.K., #22659142 to K.R.).
Conflict of interest
The authors declare that they have no conflict of interest.
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