The expression levels of miR-17-5p and HOTAIR in MSCs of patients with non-traumatic necrosis of femoral head and osteoarthritis.

<p>The MSCs were isolated from patients with non-traumatic necrosis of femoral head (ONFH, n = 15), osteoarthritis (OA, n = 13) and healthy donor (n = 10), the expression levels of miR-17-5p (A) and HOTAIR (B) were detected by real-time PCR. Each sample was repeated three times. ** <i>P</i><0.01.</p

The effects of methylation and histone acetylation alteration on miR-17-5p expression regulation.

<p>The MSCs were isolated from patients with non-traumatic necrosis of femoral head (ONFH, n = 15) and osteoarthritis (OA, n = 13). The expression levels of miR-17-5p in MSCs derived from patients with non-traumatic ONFH was detected by real-time PCR after the treatment of 5 μmol/l 5-Aza-2-deoxycytidine (5-Aza-CdR, A) or 100 nmol/l Trichostatin A (TSA, B) for 48h. The DNA methylation level of miR-17-5p promotor in MSCs from two groups were determined by bisulphite sequencing (C). Each sample was repeated three times. ** P<0.01.</p

HOTAIR regulates osteogenic differentiation markers through mediating miR-17-5p expression.

<p>Human mesenchymal stem cells from bone marrow (hMSC-BM) cell line was treated with co-transfection of siRNA-HOTAIR (si-HOTAIR) and miR-17-5p inhibitor, with si-control and NC acting as controls, respectively (A) or co-transfection of Adenovirus-HOTAIR (Ad-HOTAIR) and miR-17-5p mimic, with Ad-GFP and Pre-NC acting as controls, respectively (B) for 48h, then incubated with osteoblastic-inductive BMP-2 for six days. The mRNA levels of RUNX2 and COL1A1 were detected by real-time PCR and ALP activity was measured. n = 3, each sample was repeated three times. ** <i>P</i><0.01.</p

HOTAIR regulates osteogenic differentiation markers through mediating SMAD7 expression.

<p>Human mesenchymal stem cells from bone marrow (hMSC-BM) cell line was treated with co-transfection of siRNA-HOTAIR (si-HOTAIR) and Adenovirus-SMAD7 (Ad-SMAD7), with si-control and Ad-GFP acting as controls, respectively (A) or co-transfection of Ad-HOTAIR and si-SMAD7, with Ad-GFP and si-control acting as control, respectively (B) for 48h, then incubated with osteoblastic-inductive BMP-2 for six days. The mRNA and protein levels of RUNX2 and COL1A1 were detected by real-time PCR and western blot, and ALP activity was measured. n = 3, each sample was repeated three times. ** <i>P</i><0.01.</p

The effects of HOTAIR downregulation on the expression levels of miR-17-5p and its target gene.

<p>The MSCs were isolated from patients with non-traumatic necrosis of femoral head (ONFH), and transfected with siRNA-HOTAIR (si-HOTAIR) or siRNA-control (si-control). The HOTAIR expression level was measured by real-time PCR (A). The DNA methylation level of miR-17-5p promoter was determined by bisulphite sequencing assay (B). The miR-17-5p expression level was measured by real-time PCR (C). The SMAD7 expression in mRNA and protein levels was measured by real-time PCR and western blot (D). n = 3, each sample was repeated three times. ** <i>P</i><0.01.</p

HOTAIR expression in BMP-2-induced osteoblastic differentiation.

<p>HOTAIR expression level was detected by real-time PCR in human mesenchymal stem cells from bone marrow (hMSC-BM) cell line after the treatment of osteoblastic-inductive BMP-2 for six days. n = 3, each sample was repeated three times. ** <i>P</i><0.01.</p

The effects of HOTAIR on osteogenic differentiation markers.

<p>Human mesenchymal stem cells from bone marrow (hMSC-BM) cell line was transfected with siRNA-HOTAIR (si-HOTAIR, with si-control acting as control, A) or Adenovirus-HOTAIR (Ad-HOTAIR, with Ad-GFP acting as control, B) for 48h, then incubated with osteoblastic-inductive BMP-2 for six days. The mRNA levels of RUNX2 and COL1A1 were detected by real-time PCR and ALP activity was measured by Kits. n = 3, each sample was repeated three times. ** <i>P</i><0.01.</p

Activation of GRP78 ATPase suppresses A549 lung cancer cell migration by promoting ITGB4 degradation

Hypochlorous acid (HOCl) is an essential signal molecule in cancer cells. Activated GRP78 ATPase by a HOCl probe named ZBM-H inhibits lung cancer cell growth. However, the role and underlying mechanism of GRP78 ATPase in lung cancer cell migration have not been established. Here, we reported that activation of GRP78 ATPase by ZBM-H suppressed A549 cell migration and inhibited EMT process. Notably, ZBM-H time-dependently decreased the protein level of integrin β4 (ITGB4) in A549 cells. Combinatorial treatment of 3BDO (an autophagy inhibitor) and ZBM-H partially rescued the protein level of ITGB4. Consistently, 3BDO partially reversed ZBM-H-inhibited cell migration. Furthermore, ZBM-H promoted the interaction between ANXA7 and Hsc70, which participated in the regulation of selective autophagy and degradation of ITGB4.</p