Regulatory mechanisms and function of hypoxia-induced long noncoding RNA NDRG1-OT1 in breast cancer cells.

Hypoxia is a classic feature of the tumor microenvironment that has profound effects on cancer progression and is tightly associated with poor prognosis. Long noncoding RNAs (lncRNAs), a component of the noncoding genome, have been increasingly investigated due to their diverse roles in tumorigenesis. Previously, a hypoxia-induced lncRNA, NDRG1-OT1, was identified in MCF-7 breast cancer cells using next-generation sequencing. However, the regulatory mechanisms of NDRG1-OT1 remain elusive. Therefore, the purpose of this study was to investigate the regulatory mechanisms and functional roles of NDRG1-OT1 in breast cancer cells. Expression profiling of NDRG1-OT1 revealed that it was upregulated under hypoxia in different breast cancer cells. Overexpression and knockdown of HIF-1α up- and downregulated NDRG1-OT1, respectively. Luciferase reporter assays and chromatin immunoprecipitation assays validated that HIF-1α transcriptionally activated NDRG1-OT1 by binding to its promoter (-1773 to -1769 and -647 to -643 bp). Next, to investigate whether NDRG1-OT1 could function as a miRNA sponge, results of in silico analysis, expression profiling of predicted miRNAs, and RNA immunoprecipitation assays indicated that NDRG1-OT1 could act as a miRNA sponge of miR-875-3p. In vitro and in vivo functional assays showed that NDRG1-OT1 could promote tumor growth and migration. Lastly, a small peptide (66 a.a.) translated from NDRG1-OT1 was identified. In summary, our findings revealed novel regulatory mechanisms of NDRG1-OT1 by HIF-1α and upon miR-875-3p. Also, NDRG1-OT1 promoted the malignancy of breast cancer cells and encoded a small peptide.

Bond strength of self-adhesive resin cements to a high transparency zirconia crown and dentin.

Background/purpose: The bond strength and durability of highly translucent zirconia ceramics to dentin is still unclear. The purpose of this study was to investigate the effect of various surface treatments on the bond strength of self-adhesive resin cements to high-translucent zirconia crowns and dentin. Materials and methods: A high-transparent zirconia and three self-adhesive resin cements (G-CEM LinkAce (GCL), RelyX U200 (RXU) and TotalCem (TTC)) were used. The zirconia surface was sandblasted with 50 µm alumina particles or coated with an SR Link primer, while a dentin primer (Tetric N-Bond Universal, TBU) was applied to the surface of the dentin. By using three self-adhesive resin cements, zirconia samples were bonded to the dentin surfaces of human teeth. The shear strength of the specimens was measured before and after 10,000-cycle thermocycling or 90-day aging. Results: When using GCL to bond with the untreated dentin and various zirconia surfaces, the shear bond strength of the sandblasted (ZSB) and RS Link primer-coated (ZLK) groups was significantly higher than that of the untreated control group (Zc). However, in the case of TBU-treated dentin, the shear strength of the ZSB + LK + DTBU group was significantly higher than that of the other groups. After thermocycling and aging, the shear strength of the ZSB + LK + DTBU group using GCL and RXU cements decreased slightly, while the TTC showed no impact. Conclusion: The zirconia surface pretreated by sandblasting and bonding agent, which was sequentially bonded with a primer-treated dentin by using resin cements, can provide excellent shear bond strength and anti-aging performance.

Different effects of long noncoding RNA NDRG1-OT1 fragments on NDRG1 transcription in breast cancer cells under hypoxia.

Hypoxia plays a crucial role in the aggressiveness of solid tumors by driving multiple signaling pathways. Recently, long non-coding RNA (lncRNA) has been reported to promote or inhibit tumor aggressiveness by regulating gene expression. Previous studies in our laboratory found that the lncRNA NDRG1-OT1 is significantly up-regulated under hypoxia and inhibits its target gene NDRG1 at both the mRNA and protein levels. At the protein level, NDRG1-OT1 increases NDRG1 degradation via ubiquitin-mediated proteolysis. However, the repressive mechanism of NDRG1 at the RNA level is still unknown. Therefore, the purpose of this study was to study how NDRG1-OT1 transcriptionally regulates its target gene NDRG1. Luciferase reporter assays showed that NDRG1-OT1 decreased NDRG1 promoter activities. Mass spectrometry, bioinformatics tools, genetic manipulation, and immunoblotting were used to identify the interacting proteins. Surprisingly, different fragments of NDRG1-OT1 had opposite effects on NDRG1. The first quarter fragment (1-149 nt) of NDRG1-OT1 had no effect on the NDRG1 promoter; the second quarter fragment (150-263 nt) repressed NDRG1 by increasing the binding affinity of HNRNPA1; the third quarter fragment (264-392 nt) improved NDRG1 promoter activity by recruiting HIF-1α; the fourth quarter fragment (393-508 nt) down-regulated NDRG1 promoter activity via down-regulation of KHSRP under hypoxia. In summary, we have found a novel mechanism by which different fragments of the same lncRNA can cause opposite effects within the same target gene.