Ets-2 Regulates Cell Apoptosis via the Akt Pathway, through the Regulation of Urothelial Cancer Associated 1, a Long Non-Coding RNA, in Bladder Cancer Cells

Wenjing Wu,Mei Xue,Shuwan Zhang,Wei Chen,Xu Li,Sancheng Cao

doi:10.1371/journal.pone.0073920

Abstract

The majority of the human genome is transcribed and generates non-coding RNAs (ncRNAs) that fail to encode protein information. Long non-coding RNAs (lncRNAs) are emerging as a novel class of ncRNAs, but our knowledge about these ncRNAs is limited. Previously, our laboratory has identified that a lncRNA, Urothelial cancer associated 1 (UCA1), played an important role in bladder cancer. Despite the recent interest in UCA1 as a diagnostic marker for bladder cancer, little is known about its transcriptional regulation. To elucidate the regulation of UCA1 gene expression, we have characterized the human UCA1 gene promoter. A 2.0-kb fragment of its 5′ flanking region was cloned into a luciferase reporter vector. Deletion and mutation analysis suggested that an Ets-2 binding site was critical for UCA1 gene promoter activity. Further analysis of this site by gel shifting, chromatin immune precipitation (ChIP), and co-transfection experiments showed that transcription factor Ets-2 directly bound to the UCA1 promoter region and stimulated UCA1 promoter activity in bladder cancer cells. Taking into account the anti-apoptosis function of Ets-2, our data suggested that Ets-2 regulates apoptosis process by regulating the expression of UCA1, moreover UCA1 may be involved in the activation of Akt signaling pathway by Ets-2 in bladder cancer cells.

Highlights

Eukaryotic genomes are not the simple, well-order substrates of gene transcription that was once believed
The dysregulation of Long non-coding RNAs (lncRNAs) has been shown in various types of cancer [3,4], such as MALAT-1 in lung cancer [5], HULC in hepatocellular carcinoma [6], and HOTAIR in breast cancer [7], indicating that lncRNAs may play a critical role in tumorigenesis or tumor progression
The MegaBLAST program of NCBI was utilized to determine the transcriptional start site (TSS) of Urothelial cancer associated 1 (UCA1) gene, which was located at 15939757 position of chr19 (GRCh37/hg19)

Summary

Introduction

Eukaryotic genomes are not the simple, well-order substrates of gene transcription that was once believed. We know them to transcribe a broad spectrum of RNA molecules, ranging from long protein-coding mRNAs to short non-coding transcripts, which frequently overlap or are interleaved on either strand [1]. Long non-coding RNAs (lncRNAs) are emerging as a novel class of noncoding RNAs containing more than 200 nucleotides, but our knowledge of these lncRNAs is limited. LncRNAs are the transcripts that resemble protein-coding mRNAs in that they are capped, spliced and polyadenylated RNA polymerase II transcripts. They differ from mRNAs only in their lack of proteincoding open reading frames (ORF) [2]. In contrast to the diversity of RNA species, only a small number of lncRNAs were identified to have experimentally-derived functions. The dysregulation of lncRNA has been shown in various types of cancer [3,4], such as MALAT-1 in lung cancer [5], HULC in hepatocellular carcinoma [6], and HOTAIR in breast cancer [7], indicating that lncRNAs may play a critical role in tumorigenesis or tumor progression

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Conclusion