Journal Article

PAX8-AS1 knockdown facilitates cell growth and inactivates autophagy in osteoblasts via the miR-1252-5p/GNB1 axis in osteoporosis

Ding, Rui, Yang, Changsheng, Huang, Caiqiang, Zhang, Rongkai, Qiu, Yiyan, Li, Runguang, Xie, Denghui, Li, Qingchu

2021

Overview

Osteoporosis (OP) is the most common systematic bone disorder among elderly individuals worldwide. Long noncoding RNAs (lncRNAs) are involved in biological processes in various human diseases. It has been previously revealed that PAX8 antisense RNA 1 (PAX8-AS1) is upregulated in OP. However, its molecular mechanism in OP remains unclear. Therefore, we specifically designed this study to determine the specific role of PAX8-AS1 in OP. We first established a rat model of OP and then detected PAX8-AS1 expression in the rats with RT-qPCR. Next, to explore the biological function of PAX8-AS1 in osteoblasts, in vitro experiments, such as Cell Counting Kit-8 (CCK-8) assays, flow cytometry, western blotting and immunofluorescence (IF) staining, were conducted. Subsequently, we performed bioinformatic analysis and luciferase reporter assays to predict and identify the relationships between microRNA 1252-5p (miR-1252-5p) and both PAX8-AS1 and G protein subunit beta 1 (GNB1). Additionally, rescue assays in osteoblasts clarified the regulatory network of the PAX8-AS1/miR-1252-5p/GNB1 axis. Finally, in vivo loss-of-function studies verified the role of PAX8-AS1 in OP progression. The results illustrated that PAX8-AS1 was upregulated in the proximal tibia of OP rats. PAX8-AS1 silencing promoted the viability and inhibited the apoptosis and autophagy of osteoblasts. PAX8-AS1 interacted with miR-1252-5p. GNB1 was negatively regulated by miR-1252-5p. In addition, the impacts of PAX8-AS1 knockdown on osteoblasts were counteracted by GNB1 overexpression. PAX8-AS1 depletion suppressed OP progression by inhibiting apoptosis and autophagy in osteoblasts. In summary, PAX8-AS1 suppressed the viability and activated the autophagy of osteoblasts via the miR-1252-5p/GNB1 axis in OP. Osteoporosis: An RNA molecule that suppresses bone-building A recently identified regulatory RNA molecule contributes to osteoporosis by suppressing production of a protein that promotes survival of bone-building osteoblast cells. These regulatory RNA molecules do not encode proteins, but instead regulate gene expression by binding and inactivating complementary RNA strands. Researchers led by Yiyan Qiu at Southern Medical University in Guangzhou, China, have now learned how one such RNA facilitates bone degeneration in osteoporosis. Qiu and colleagues show that this RNA interferes with the production of a protein that is essential to the metabolic health and survival of osteoblasts, promoting bone loss in a rat model of osteoporosis. Experimental techniques that selectively deplete this RNA help preserve the viability of the osteoblast population. This depletion could provide a useful approach for future drug development efforts.

• Publisher: Nature Publishing Group UK,Springer Nature B.V,생화학분자생물학회
• Subject: 13/1 / 631/80/84 / 692/699/2768 / 82/80 / Animals / Antisense RNA / Antisense therapy / Apoptosis / Apoptosis - genetics / Autophagy / Autophagy - genetics / Biomedical and Life Sciences / Biomedicine / Bone loss / Cell Line, Tumor / Cholecystokinin / Degeneration / Disease Models, Animal / Drug development / Female / Flow cytometry / Gene expression / Gene Knockdown Techniques / Gene Silencing / GTP-Binding Protein beta Subunits - genetics / Immunofluorescence / Medical Biochemistry / MicroRNAs - genetics / miRNA / Models, Biological / Molecular Medicine / Osteoblasts / Osteoblasts - cytology / Osteoblasts - metabolism / Osteoporosis / Osteoporosis - etiology / Osteoporosis - metabolism / Osteoporosis - pathology / Pax8 protein / PAX8 Transcription Factor - genetics / Phagocytosis / Proteins / Rats / Ribonucleic acid / RNA / RNA Interference / RNA, Antisense - genetics / RNA, Long Noncoding - genetics / Stem Cells / Tibia / Viability / Western blotting / 생화학