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Urothelial carcinoma associated 1 is a hypoxia-inducible factor-1α-targeted long noncoding RNA that enhances hypoxic bladder cancer cell proliferation, migration, and invasion

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Tumor Biology

Abstract

Urothelial carcinoma associated 1 (UCA1) has been identified as an oncogenic long noncoding RNA (lncRNA) that is involved in bladder cancer progression and acts as a diagnostic biomarker for bladder carcinoma. Here, we studied the expression and function of lncRNA-UCA1 in the hypoxic microenvironment of bladder cancer. The expression and transcriptional activity of lncRNA-UCA1 were measured by quantitative real-time polymerase chain reaction and luciferase assays. Cell proliferation and apoptosis were evaluated by MTT assays and flow cytometry. Cell migration and invasion were detected by wound healing, migration, and invasion assays. The binding of hypoxia-inducible factor-1α (HIF-1α) to hypoxia response elements (HREs) in the lncRNA-UCA1 promoter was confirmed by electrophoretic mobility shift assay and chromatin immunoprecipitation. HRE mutations were generated by using a site-directed mutagenesis kit, and HIF-1α knockdown was mediated by small interfering RNA. The effect of HIF-1α inhibition by YC-1 on lncRNA-UCA1 expression was also examined. LncRNA-UCA1 was upregulated by hypoxia in bladder cancer cells. Under hypoxic conditions, lncRNA-UCA1 upregulation increased cell proliferation, migration, and invasion and inhibited apoptosis. The underlying mechanism of hypoxia-upregulated lncRNA-UCA1 expression was that HIF-1α specifically bound to HREs in the lncRNA-UCA1 promoter. Furthermore, HIF-1α knockdown or inhibition could prevent lncRNA-UCA1 upregulation under hypoxia. These findings revealed the mechanism of lncRNA-UCA1 upregulation in hypoxic bladder cancer cells and suggested that effective blocking of lncRNA-UCA1 expression in the hypoxic microenvironment of bladder cancer could be a novel therapeutic strategy.

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Acknowledgments

This work was supported by the grants (No. 81072104) from the National Natural Science Foundation of China.

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Authors and Affiliations

  1. Center for Translational Medicine, The First Affiliated Hospital, School of Medicine, Xi’an Jiaotong University, Xi’an, Peoples Republic of China

    Mei Xue, Xu Li & Zhengkun Li

  2. Clinical Laboratory, The First Affiliated Hospital, School of Medicine, Xi’an Jiaotong University, 277 West Yanta Road, 710061, Xi’an, Peoples Republic of China

    Wei Chen

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  1. Mei Xue
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Correspondence to Wei Chen.

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Supplementary Figure 1

a lncRNA-UCA1 expression levels were analyzed by PCR in SKOV3 cells under both normoxia and hypoxia. 18S rRNA was used as the internal control (*P < 0.05, n = 3). Line 1-2 indicated normoxic SKOV3 cells; line 3-4 indicated hypoxic SKOV3 cells. b ChIP showed the interaction of HIF-1α with the lncRNA-UCA1 promoter in hypoxic SKOV3 cells (*P < 0.05, n = 3) (GIF 37 kb)

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Xue, M., Li, X., Li, Z. et al. Urothelial carcinoma associated 1 is a hypoxia-inducible factor-1α-targeted long noncoding RNA that enhances hypoxic bladder cancer cell proliferation, migration, and invasion. Tumor Biol. 35, 6901–6912 (2014). https://doi.org/10.1007/s13277-014-1925-x

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  • DOI: https://doi.org/10.1007/s13277-014-1925-x

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