LncRNA HOTAIR is a novel endothelial mechanosensitive gene

Abstract

To better address whether the long noncoding RNAs (lncRNAs) HOTAIR and HOTTIP are mechanosensitive genes, they were investigated in differentially challenged endothelial cells with respect to a circuit of tensional forces, considering the performance of both arterial and venous endothelial cells. We subjected arterial-and venous-obtained endothelial cells to a circuit of tensional forces within a shear stress model in vitro. Real-time quantitative polymerase chain reaction analysis indicated that microRNA (miRNA)-related processing machinery is significantly required in shear stressed arterial endothelial cell metabolism, which orchestrates miRNA (small noncoding RNA) involvement, and their involvement suggests lncRNA involvement. Of lncRNAs HOTAIR and HOTTIP, only HOTAIR was mechanosensitive considering both arterial and venous endothelial cells, presenting a positive correlation between methylation signature and gene expression. Thereafter, using bioinformatics tools, lncRNA HOTAIR was predicted to modulate miRNA185, miRNA-21, and miRNA23b downregulation. We compared the values of gene expression with a Pearson's correlation test, and expected correlations were observed for miRNA185 (r = 0.8664), miRNA-21 (r = 0.8605), and miRNA23b (0.9128). Taken together, these findings clearly show that lncRNA HOTAIR responds to the shear stress and emerges as a novel mechanosensitive gene in endothelial cells. Altogether, this understanding of mechanosensitive transcriptional and posttranscriptional control involving HOTAIR can also lead to new forms of therapeutic intervention for various diseases, as well as new strategies for tissue engineering and regenerative medicine.

Epigenetic Differences Arise in Endothelial Cells Responding to Cobalt–Chromium

“…Epigenetic mechanisms control the levels of gene expression and the possible phenotype changes in cells [ 32 , 41 ]. Specifically, epigenetic metabolism is performed by the mechanisms of acetylation, methylation, micro-RNA (miRNAs), and long non-coding RNA (lnc-RNAs) controls [ 42 , 43 , 44 , 45 , 46 ]. As angiogenesis is widely known to be decisive during bone oppositional growth during the osseointegration of biomaterials in dentistry and medicine, the goal here was to better address the epigenetic mechanism, which is able to drive the phenotype of endothelial cells responding to Co-Cr.…”

Nanohydroxyapatite-Blasted Bioactive Surface Drives Shear-Stressed Endothelial Cell Growth and Angiogenesis

“…HUVECs were seeded in the peripheral area of the modified 100 mm culture dishes to allow the application of shear stress methodology. To note, the modified 100 mm culture dishes were obtained by bonding at the bottom a 60 mm culture dishes into their center using medical silicone, and thereafter, the dishes were sterilized using UV light for 30 min [ 20 , 23 – 25 ] ( Figure 1 ).…”

“…HUVECs were previously seeded onto the peripheral area of the modified culture dishes (as described earlier) and then subjected to the orbital shear stress-induced tension force, using an orbital shaker (Scilogex, Rocky Hill, CT) positioned inside of the cell culture incubator, at 37°C in the presence of CO 2 and 95% humidity, as proposed by others [ 19 , 20 , 23 , 26 – 28 ]. The shear stress model performed here uses an orbital shaker respecting the formula of the maximal wall shear stress: τ max = α √ ρη (2 πʄ )3, in which τ max (Pascal unit) is the shear stress, α is the radius of orbital rotation (12 cm), ρ is the density of the cell culture medium (937.5 kg/m 3 ), η is the viscosity of the cell culture medium (7.5 × 10 −4 Pa s), and ʄ is the frequency of rotation.…”

Combination of in silico and cell culture strategies to predict biomaterial performance: Effects of sintering temperature on the biological properties of hydroxyapatite

“…To evaluate the molecular mechanism related with the adaptation to each HA, the osteoblasts were newly subjected to each HAconditioned medium up to 48 h, when the cells were harvested and total RNA extracted using Ambion TriZOL Reagent (Life Sciences-Fisher Scientific Inc., Waltham, MA, USA) and treated with DNAse I (Invitrogen, Carlsband, CA, USA), as used and described earlier. 29 According to the manufacturer's instructions, cDNA synthesis was performed with the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA, USA). Real-time PCR was performed on 10 μL containing PowerUpTM SYBR™ Green Master Mix 2Â (5 μL) (Applied Biosystems, Foster City, CA, USA), 0.4 μM of each primer, 50 ng of cDNA and H 2 O nuclease-free.…”