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. 2020 Jan;19(1):153-159.
doi: 10.3892/etm.2019.8185. Epub 2019 Nov 12.

LncRNA AWPPH participates in the development of non-traumatic osteonecrosis of femoral head by upregulating Runx2

Xiantao Chen  1 , Jing Li  1 , Dawei Liang  1 , Leilei Zhang  1 , Qingfeng Wang  1
Affiliations

LncRNA AWPPH participates in the development of non-traumatic osteonecrosis of femoral head by upregulating Runx2

Xiantao Chen et al. Exp Ther Med. 2020 Jan.

Abstract

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.

Keywords: Runx2; long noncoding RNA AWPPH; mesenchymal stem cells; non-traumatic osteonecrosis of femoral head.

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Figures

Figure 1.
Figure 1.
Expression of lncRNA AWPPH and Runx2 mRNA in MSCs and serum derived from non-traumatic ONFH patients and healthy people. This figure presents the expression of lncRNA AWPPH in (A) MSCs and (B) serum and the expression of Runx2 mRNA in (C) MSCs and (D) serum collected from patients with non-traumatic ONFH and healthy people. *P<0.05. MSCs, mesenchymal stem cells; Runx2, runt-related transcription factor 2; lnc, long noncoding; ONFH, osteonecrosis of femoral head.
Figure 2.
Figure 2.
Diagnostic value of lncRNA AWPPH expression in MSCs and serum for patients with non-traumatic ONFH. This figure demonstrated the receiver operating characteristics curve of the uses of AWPPH expression in (A) MSCs and (B) serum for the diagnosis of non-traumatic ONFH. MSCs, mesenchymal stem cells; lnc, long noncoding; ONFH, osteonecrosis of femoral head.
Figure 3.
Figure 3.
Expression of lncRNA AWPPH in hMSC-BM cells under treatment of different concentrations of BMP-2 (0, 25, 100 and 200 ng/ml). *P<0.05. hMSCs-BM, human mesenchymal stem cells-bone marrow; lnc, long noncoding; BMP-2 bone morphogenic protein-2.
Figure 4.
Figure 4.
Effects of AWPPH overexpression and shRNA silencing on expression of Runx2 in hMSC-BM cells. *P<0.05; Negative control cells transfected with empty vector (left) or negative control shRNA (right). This figure exhibits the effects of AWPPH overexpression and shRNA silencing on expression of (A) AWPPH and (B) Runx2 in hMSC-BM cells. Sh, short harpin; hMSCs-BM, human mesenchymal stem cells-bone marrow; Runx2, runt-related transcription factor 2.
See this image and copyright information in PMC

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