Effect ofPeriostinSilencing on the Autophagy of Osteoblasts

Abstract

The objective of the present work was to investigate the effect of Periostin (POSTN) silencing on autophagy in osteoblasts, and provide an experimental basis for studying the mechanism of dental eruption. The cells were divided into the following four groups according to their viral number: the NC group, pFU-GW-016PSC53349-1; group KD1, LVpFU-GW-016PSC66471-1; group KD2, LVpFU-GW-016PSC66472-1; and group KD3, LVpFU-GW-016PSC66473-1. The lentiviral vector was infected at MOI = 100 in the ENi.S medium containing 5 g/mL Polybrene. The target gene expression was observed by a Celigo Ò Image Cytometer at 72 hours after infection, and the positive rate of fluorescence was noted. A two-step method of quantitative real-time PCR (qRT-PCR) was used to detect the silencing effect of POSTN. Western blotting was then performed to assess the expression of autophagy-related proteins Beclin-1 and LC3 in the group showing the best gene silencing effects. The experimental results showed that there was strong green fluorescence in group KD3. As confirmed via qRT-PCR analysis, the POSTN silencing efficiency in group KD3 reached 92.1%. The Western blotting revealed that the expression of Beclin-1 protein in group KD3 was significantly higher than that in the NC group. However, the LC3 protein expression was not significantly different from that of the control group. The lentiviral vector targeting POSTN in osteoblasts was constructed successfully. In addition, the expression of autophagy protein in mouse osteoblasts increased after POSTN silencing. This finding may provide new approaches for understanding the molecular signal transduction of POSTN during the tooth eruption process.

Periostin regulates autophagy through integrin α5β1 or α6β4 and an AKT‐dependent pathway in colorectal cancer cell migration

“…In CRC, cell autonomy and non‐autonomous roles for autophagy are essential in growth and progression; however, the mechanisms downstream of autophagy to reduce or enhance tumour growth are not well known 15 . It was recently reported that autophagy protein expression levels in mouse osteoblasts were increased after PN silencing 16 . Up to present knowledge, the alterations of autophagy on CRC tumour promotion after exposure to PN are still unclear.…”

“…15 It was recently reported that autophagy protein expression levels in mouse osteoblasts were increased after PN silencing. 16 Up to present knowledge, the alterations of autophagy on CRC tumour promotion after exposure to PN are still unclear.…”

Autophagy mediates cementoblast mineralization through periostin/β-catenin signaling axis under compression

“…In addition, decreased levels of LC3 have been observed following exposure to Postn 21 . The autophagic function of osteoblasts is enhanced when Postn is knocked down 22 . Consistent with previous studies, we demonstrated that Postn knockdown enhanced autophagic activity in cementoblasts.…”

“…Collectively our data indicate that knockdown of Postn inhibits cementoblast mineralization in vitro. Growing evidence suggests that Postn has a regulatory effect on autophagy 21,22 ; thus, we examined the effects of Postn on autophagy in cementoblasts. Autophagy was upregulated with knockdown of Postn, as re ected by decreased levels of P62 and increased conversion of LC3-I to LC3-II (Fig.…”

Periostin: An Emerging Molecule With a Potential Role in Spinal Degenerative Diseases

“…The results showed that the expression of the autophagy-related protein beclin 1 in osteoblasts was increased, suggesting that the autophagy function of osteoblasts was enhanced. This also indicates that POSTN inhibits autophagy in osteoblasts, although the specific signaling pathway mediating this effect has not yet been identified (83). In a subsequent study, these authors demonstrated that the protein expression of RUNX2 and RANKL was significantly reduced after POSTN gene silencing, and speculated that this process was achieved through the NF-κB signaling pathway (84).…”