Department of Endocrinology, The First People's Hospital of Fuyang District, Hangzhou, Zhejiang, China.
PLoS One. 2021 May 27;16(5):e0252126. doi: 10.1371/journal.pone.0252126. eCollection 2021.
Osteoporosis affects the quality of life among middle-aged and elderly individuals. In addition, dysfunction of osteoblasts can lead to the progression of osteoporosis. Circular (circ)RNAs are involved in various types of diseases, including osteoporosis. Moreover, it has been reported that hsa_circ_0001275 expression is upregulated in osteoporosis. However, the effects of hsa_circ_0001275 on the growth of osteoblasts remain unclear.
In the present study, the gene and protein expression levels in hFOB1.19 cells were detected via reverse transcription-quantitative (RT-qPCR) and western blot analyses, respectively. In addition, alkaline phosphatase (ALP) activity and calcium nodules were examined by ALP and alizarin red staining, respectively. Cell proliferation was measured using the Cell Counting Kit-8 assay. Cell apoptosis and cell cycle were analyzed by flow cytometry. Furthermore, dual luciferase reporter and RNA pull-down assay were used to confirm the association among hsa_circ_0001275, microRNA (miR)-377 and CDKN1B.
DEX-induced hFOB1.19 cell growth inhibition was significantly reversed by silencing hsa_circ_0001275. Moreover, DEX significantly increased ALP activity and calcium nodules in hFOB1.19 cells, while this effect was significantly reversed in the presence of hsa_circ_0001275 small interfering RNA. In addition, miR-377 was sponged by hsa_circ_0001275 and CDKN1B was directly targeted by miR-377 in hFOB1.19 cells. Furthermore, the therapeutic effect of hsa_circ_0001275 knockdown on osteoporosis was notably reversed by miR-377 antagomir.
The data demonstrated that knockdown of hsa_circ_0001275 reversed DEX-induced osteoblast growth inhibition via activation of the miR-377/CDKN1B axis. Therefore, this study might shed new lights on the treatment of osteoporosis.
骨质疏松症会影响中老年人的生活质量。此外，成骨细胞功能障碍可导致骨质疏松症的进展。环状（circ）RNAs 参与多种疾病，包括骨质疏松症。此外，据报道 hsa_circ_0001275 在骨质疏松症中表达上调。然而，hsa_circ_0001275 对成骨细胞生长的影响尚不清楚。
在本研究中，通过逆转录定量（RT-qPCR）和 Western blot 分析分别检测 hFOB1.19 细胞中的基因和蛋白表达水平。此外，通过碱性磷酸酶（ALP）和茜素红染色分别检测 ALP 活性和钙结节。通过细胞计数试剂盒-8 测定法测量细胞增殖。通过流式细胞术分析细胞凋亡和细胞周期。此外，使用双荧光素酶报告和 RNA 下拉测定来验证 hsa_circ_0001275、微小 RNA（miR）-377 和 CDKN1B 之间的关联。
沉默 hsa_circ_0001275 显著逆转了 DEX 诱导的 hFOB1.19 细胞生长抑制。此外，DEX 显著增加了 hFOB1.19 细胞中的 ALP 活性和钙结节，而在存在 hsa_circ_0001275 小干扰 RNA 的情况下，这种作用显著逆转。此外，miR-377 被 hsa_circ_0001275 海绵吸附，CDKN1B 是 hFOB1.19 细胞中 miR-377 的直接靶标。此外，hsa_circ_0001275 敲低对骨质疏松症的治疗作用在 miR-377 拮抗剂存在的情况下明显逆转。
数据表明，沉默 hsa_circ_0001275 通过激活 miR-377/CDKN1B 轴逆转了 DEX 诱导的成骨细胞生长抑制。因此，本研究可能为骨质疏松症的治疗提供新的思路。
