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Upregulation of long non-coding RNA urothelial carcinoma associated 1 by CCAAT/enhancer binding protein α contributes to bladder cancer cell growth and reduced apoptosis.

Xue M, Li X, Wu W, Zhang S, Wu S, Li Z, Chen W - Oncol. Rep. (2014)

Bottom Line: We also demonstrated that C/EBPα siRNA treatment contributed to the downregulation of lncRNA-UCA1 expression, whereas overexpression of C/EBPα enhanced lncRNA-UCA1 expression.Furthermore, lncRNA-UCA1 transcriptional repression by C/EBPα siRNA sharply reduced cell viability and induced cell apoptosis in vitro.Collectively, our results provide a novel therapeutic strategy for bladder cancer by effectively interrupting the binding of the lncRNA-UCA1 promoter and certain transcription factors, so as to reverse the upregulation of lncRNA-UCA1 and prevent bladder cancer progression.

View Article: PubMed Central - PubMed

Affiliation: Center for Translational Medicine, The First Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an 710061, P.R. China.

ABSTRACT
Long non-coding RNA urothelial carcinoma associated 1 (lncRNA-UCA1) is upregulated in bladder cancer and plays a pivotal role in bladder cancer progression and metastasis. Recent studies and our research found that lncRNA-UCA1 may be an important biomarker and therapeutic target for bladder cancer. However, the molecular mechanism involved in the upregulation of lncRNA-UCA1 in bladder cancer is largely unknown. In the present study, we showed that lncRNA-UCA1 expression in bladder cancer cells was upregulated by transcription factor CCAAT/enhancer binding protein α (C/EBPα), which was the only candidate transcription factor simultaneously predicted by a total of five bioinformatical software programs. Electrophoretic mobility shift assay and chromatin immunoprecipitation assay indicated that C/EBPα bound to the lncRNA-UCA1 core promoter region in vitro and in vivo. The luciferase assays further showed that there was a point mutation (A231G) in the C/EBPα binding site of the lncRNA-UCA1 core promoter in various bladder cancer cell lines, which in turn significantly increased the transcriptional activity of lncRNA-UCA1. We also demonstrated that C/EBPα siRNA treatment contributed to the downregulation of lncRNA-UCA1 expression, whereas overexpression of C/EBPα enhanced lncRNA-UCA1 expression. Furthermore, lncRNA-UCA1 transcriptional repression by C/EBPα siRNA sharply reduced cell viability and induced cell apoptosis in vitro. Collectively, our results provide a novel therapeutic strategy for bladder cancer by effectively interrupting the binding of the lncRNA-UCA1 promoter and certain transcription factors, so as to reverse the upregulation of lncRNA-UCA1 and prevent bladder cancer progression.

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Bioinformatic analysis of the lncRNA-UCA1 core promoter region. (A) Schematic representation of the putative C/EBPα binding site in the lncRNA-UCA1 core promoter. (B) Sequence logo of C/EBPα was obtained from the JASPAR database. (C) C/EBPα and lncRNA-UCA1 expression levels were analyzed in the various cell lines by real-time PCR. β-actin was used as the internal control.
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f1-or-31-05-1993: Bioinformatic analysis of the lncRNA-UCA1 core promoter region. (A) Schematic representation of the putative C/EBPα binding site in the lncRNA-UCA1 core promoter. (B) Sequence logo of C/EBPα was obtained from the JASPAR database. (C) C/EBPα and lncRNA-UCA1 expression levels were analyzed in the various cell lines by real-time PCR. β-actin was used as the internal control.

Mentions: In our previous study, we confirmed that the lncRNA-UCA1 promoter was located at the 5′ end of the lncRNA-UCA1 gene, from −1800 bp to +200 bp, with the core promoter ranging from −400 bp to −150 bp (17). Several potential transcription factor binding sites were predicted in the lncRNA-UCA1 core promoter region. C/EBPα was the only candidate transcription factor predicted by five bioinformatical software programs simultaneously (Table II). It was further speculated that there were more than one putative C/EBPα binding site in the lncRNA-UCA1 core promoter region (Table III). Based on the JASPAR database, we determined that a unique motif of C/EBPα (from −239 bp to −230 bp, GTTTCCAAA) was potentially eligible to interact and bind with the lncRNA-UCA1 core promoter (Fig. 1A and B). In order to screen out a perfect cell model by which to validate our prediction results, we detected the constitutive expression of C/EBPα and lncRNA-UCA1 in three bladder cancer cell lines. As shown in Fig. 1C, C/EBPα was expressed in all of the three bladder cell lines. lncRNA-UCA1 expression was high in the cell line BLZ-211, yet extremely low, if not absent, in its counterpart cell line BLS-211, although the two cell lines were derived from the same patient (16). Therefore, BLS-211 was employed as the control cell line in the subsequent experiments.


Upregulation of long non-coding RNA urothelial carcinoma associated 1 by CCAAT/enhancer binding protein α contributes to bladder cancer cell growth and reduced apoptosis.

Xue M, Li X, Wu W, Zhang S, Wu S, Li Z, Chen W - Oncol. Rep. (2014)

Bioinformatic analysis of the lncRNA-UCA1 core promoter region. (A) Schematic representation of the putative C/EBPα binding site in the lncRNA-UCA1 core promoter. (B) Sequence logo of C/EBPα was obtained from the JASPAR database. (C) C/EBPα and lncRNA-UCA1 expression levels were analyzed in the various cell lines by real-time PCR. β-actin was used as the internal control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4020618&req=5

f1-or-31-05-1993: Bioinformatic analysis of the lncRNA-UCA1 core promoter region. (A) Schematic representation of the putative C/EBPα binding site in the lncRNA-UCA1 core promoter. (B) Sequence logo of C/EBPα was obtained from the JASPAR database. (C) C/EBPα and lncRNA-UCA1 expression levels were analyzed in the various cell lines by real-time PCR. β-actin was used as the internal control.
Mentions: In our previous study, we confirmed that the lncRNA-UCA1 promoter was located at the 5′ end of the lncRNA-UCA1 gene, from −1800 bp to +200 bp, with the core promoter ranging from −400 bp to −150 bp (17). Several potential transcription factor binding sites were predicted in the lncRNA-UCA1 core promoter region. C/EBPα was the only candidate transcription factor predicted by five bioinformatical software programs simultaneously (Table II). It was further speculated that there were more than one putative C/EBPα binding site in the lncRNA-UCA1 core promoter region (Table III). Based on the JASPAR database, we determined that a unique motif of C/EBPα (from −239 bp to −230 bp, GTTTCCAAA) was potentially eligible to interact and bind with the lncRNA-UCA1 core promoter (Fig. 1A and B). In order to screen out a perfect cell model by which to validate our prediction results, we detected the constitutive expression of C/EBPα and lncRNA-UCA1 in three bladder cancer cell lines. As shown in Fig. 1C, C/EBPα was expressed in all of the three bladder cell lines. lncRNA-UCA1 expression was high in the cell line BLZ-211, yet extremely low, if not absent, in its counterpart cell line BLS-211, although the two cell lines were derived from the same patient (16). Therefore, BLS-211 was employed as the control cell line in the subsequent experiments.

Bottom Line: We also demonstrated that C/EBPα siRNA treatment contributed to the downregulation of lncRNA-UCA1 expression, whereas overexpression of C/EBPα enhanced lncRNA-UCA1 expression.Furthermore, lncRNA-UCA1 transcriptional repression by C/EBPα siRNA sharply reduced cell viability and induced cell apoptosis in vitro.Collectively, our results provide a novel therapeutic strategy for bladder cancer by effectively interrupting the binding of the lncRNA-UCA1 promoter and certain transcription factors, so as to reverse the upregulation of lncRNA-UCA1 and prevent bladder cancer progression.

View Article: PubMed Central - PubMed

Affiliation: Center for Translational Medicine, The First Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an 710061, P.R. China.

ABSTRACT
Long non-coding RNA urothelial carcinoma associated 1 (lncRNA-UCA1) is upregulated in bladder cancer and plays a pivotal role in bladder cancer progression and metastasis. Recent studies and our research found that lncRNA-UCA1 may be an important biomarker and therapeutic target for bladder cancer. However, the molecular mechanism involved in the upregulation of lncRNA-UCA1 in bladder cancer is largely unknown. In the present study, we showed that lncRNA-UCA1 expression in bladder cancer cells was upregulated by transcription factor CCAAT/enhancer binding protein α (C/EBPα), which was the only candidate transcription factor simultaneously predicted by a total of five bioinformatical software programs. Electrophoretic mobility shift assay and chromatin immunoprecipitation assay indicated that C/EBPα bound to the lncRNA-UCA1 core promoter region in vitro and in vivo. The luciferase assays further showed that there was a point mutation (A231G) in the C/EBPα binding site of the lncRNA-UCA1 core promoter in various bladder cancer cell lines, which in turn significantly increased the transcriptional activity of lncRNA-UCA1. We also demonstrated that C/EBPα siRNA treatment contributed to the downregulation of lncRNA-UCA1 expression, whereas overexpression of C/EBPα enhanced lncRNA-UCA1 expression. Furthermore, lncRNA-UCA1 transcriptional repression by C/EBPα siRNA sharply reduced cell viability and induced cell apoptosis in vitro. Collectively, our results provide a novel therapeutic strategy for bladder cancer by effectively interrupting the binding of the lncRNA-UCA1 promoter and certain transcription factors, so as to reverse the upregulation of lncRNA-UCA1 and prevent bladder cancer progression.

Show MeSH
Related in: MedlinePlus