Chen Xiantao, Li Jing, Liang Dawei, Zhang Leilei, Wang Qingfeng
Department of Osteonecrosis of The Femoral Head, Luoyang Orthopedic Hospital of Henan Province, Luoyang, Henan 471000, P.R. China.
Exp Ther Med. 2020 Jan;19(1):153-159. doi: 10.3892/etm.2019.8185. Epub 2019 Nov 12.
AWPPH is a newly discovered long noncoding (lnc)RNA that plays an oncogenic role in development of several types of malignancies, whiles its involvement in non-traumatic osteonecrosis of femoral head (ONFH) is unknown. Therefore, the present study aimed to investigate the functionality of AWPPH in non-traumatic ONFH. Blood and mesenchymal stem cells (MSCs) were obtained from both non-traumatic ONFH patients and healthy controls, and expression of AWPPH in those tissues was detected by RT-qPCR. Receiver operating characteristic curve analysis was performed to investigate the diagnostic value of lncRNA AWPPH expression for non-traumatic ONFH. Bone morphogenic protein (BMP-2) was used to treat MSCs to induce osteogenic differentiation and the effects on lncRNA AWPPH expression was detected by RT-qPCR. LncRNA AWPPH overexpression and short hairpin (sh)RNA silencing cell lines were established and the effects on runt-related transcription factor 2 (Runx2) expression were detected by western blotting. It was demonstrated that AWPPH was significantly downregulated in non-traumatic ONFH patients compared with in healthy controls in both MSCs and serum. Expression of AWPPH in MSCs and serum is a sensitive diagnostic marker for non-traumatic ONFH. Expression of AWPPH exhibited no significant correlation with patients' age, gender and living habits, but was significantly correlated with course of disease. BMP-2 treatment significantly increased the expression level of AWPPH in human MSCs from bone marrow (hMSC-BM). AWPPH overexpression promoted, while AWPPH short hairpin RNA silencing inhibited the expression of Runx2 expression in hMSC-BM cells. Therefore, it was concluded that lncRNA AWPPH may participate in the development of ONFH by upregulating Runx2.
AWPPH是一种新发现的长链非编码（lnc）RNA，在多种恶性肿瘤的发生发展中发挥致癌作用，但其在非创伤性股骨头坏死（ONFH）中的作用尚不清楚。因此，本研究旨在探讨AWPPH在非创伤性ONFH中的功能。从非创伤性ONFH患者和健康对照者中获取血液和间充质干细胞（MSC），通过RT-qPCR检测这些组织中AWPPH的表达。进行受试者工作特征曲线分析，以研究lncRNA AWPPH表达对非创伤性ONFH的诊断价值。用骨形态发生蛋白（BMP-2）处理MSC以诱导成骨分化，并通过RT-qPCR检测对lncRNA AWPPH表达的影响。建立lncRNA AWPPH过表达和短发夹（sh）RNA沉默细胞系，并通过蛋白质免疫印迹法检测对 runt相关转录因子2（Runx2）表达的影响。结果表明，与健康对照者相比，非创伤性ONFH患者的MSC和血清中AWPPH均显著下调。MSC和血清中AWPPH的表达是诊断非创伤性ONFH的敏感标志物。AWPPH的表达与患者的年龄、性别和生活习惯无显著相关性，但与病程显著相关。BMP-2处理显著提高了人骨髓间充质干细胞（hMSC-BM）中AWPPH的表达水平。AWPPH过表达促进了hMSC-BM细胞中Runx2的表达，而AWPPH短发夹RNA沉默则抑制了Runx2的表达。因此，得出结论：lncRNA AWPPH可能通过上调Runx2参与ONFH的发生发展。
