Long Noncoding RNA HAGLROS Promotes Cell Invasion and Metastasis by Sponging miR-152 and Upregulating ROCK1 Expression in Osteosarcoma.

BACKGROUND: Long noncoding RNAs (lncRNAs) played a crucial role in a number of biological processes. lncRNA HAGLROS was demonstrated to facilitate cell proliferation and migration in various cancers. However, the functions and molecular mechanisms of HAGLROS in osteosarcoma remained to be elucidated. METHODS: qRT-PCR assay was used to detect the relative expression of HAGLROS in osteosarcoma tissue samples and cells. CCK-8 and Transwell assays were performed to assess the effects of HAGLROS on OS cells proliferation and invasion. Luciferase reporter assay verified the interaction between ROCK1 and miR-152. RESULTS: In our study, we found that the expression of HAGLROS increased osteosarcoma samples and cell lines compared with normal tissues and cells. HAGLROS knockdown inhibited certain functions of U2OS and SW1353 cells in vitro. Moreover, HAGLROS depletion inhibited tumor growth and metastasis in vivo. Mechanically, we found that HAGLROS sponged miR-152 to promote ROCK1 expression in U2OS and SW1353 cells. CONCLUSION: In summary, our study indicated that HAGLROS could promote osteosarcoma progression by sponging miR-152 to promote ROCK1 expression. The results showed HAGLROS/miR-152/ROCK1 axis might act as a novel therapeutic strategy for osteosarcoma.

LINC00511 Promotes Osteosarcoma Tumorigenesis and Invasiveness through the miR-185-3p/E2F1 Axis.

Osteosarcoma is a malignant tumor that seriously threatens human health. Numerous studies have pointed out the potential of long noncoding RNAs (lncRNAs) as new therapeutic targets for various human cancers. Therefore, we mainly investigate whether there is a new type of lncRNA pathway involved in regulating the development of osteosarcoma. The present study shows the higher expression levels of LINC00511 correlates to a shorter overall survival and disease-free survival time in patients with sarcoma. It is significantly higher in the clinical samples of osteosarcoma patients than in normal adjacent cancer tissues. We used U373 and SW1353 osteosarcoma cells to determine the effect of lncRNA on osteosarcoma proliferation and invasion by knocking down LINC00511 compared with controls. The results showed that the LINC00511 knockdown significantly suppressed osteosarcoma cell growth and metastasis. To explore the mechanisms of LINC00511 in osteosarcoma, we tested whether LINC00511 could competitively stimulate miR-185-3p and regulate E2F1 as a ceRNA. The results showed that LINC00511 knockdown induced the increased level of miR-185-3p levels; however, miR-185-3p overexpression suppressed LINC00511 levels. In addition, the results also demonstrated that LINC00511 knockdown or miR-185-3p overexpression could reduce E2F1 levels in osteosarcoma cells. The dual-luciferase reporter assay verified the direct interaction between miR-185-3p and LINC00511 or E2F1. These results may offer an explanation of how the lncRNA affects the progression of osteosarcoma, and our study shows that LINC00511 can be a novel biomarker in osteosarcoma.

Circular RNA circHIPK3 Promotes Cell Metastasis through miR-637/STAT3 Axis in Osteosarcoma.

Recent studies have suggested that circular RNAs play an important role in the progression of various cancers. However, few studies have revealed the great value of circRNAs in the diagnosis and prognosis prediction of osteosarcoma (OS). In this study, we performed experiments with the human OS cell lines and the results showed that the expression of circHIPK3 in OS cell lines was significantly upregulated compared to that in the normal cell line. In addition, the results showed that circHIPK3 could promote the migration, invasion, and growth of OS cells. Furthermore, miR-637 was identified as a target of circHIPK3, while STAT3 was targeted by miR-637. circHIPK3 could promote STAT3 expression via interacting with miR-637 in OS cells. In conclusion, our research uncovered an important role of the circHIPK3/miR-637/STAT3 pathway in the migration and invasion of OS cells and suggested that circHIPK3 may be a prognostic marker and a promising therapeutic target for OS.

Identification of differentially expressed microRNAs in the bone marrow of osteoporosis patients.

This study aimed to identify specific microRNAs (miRNAs) related to postmenopausal osteoporosis (OP) in human. A total of 67 conserved miRNAs, including 50 miRNAs significantly up-regulated and 17 miRNAs significantly downregulated, showed differential expression between OP group and control group. 180 hairpin structures were predicted and 199 potential novel miRNA candidates with 18 to 25 nt in length, which will greatly enrich the human miRBase. 4 miRNAs (miR-518b, miR-582-3p, miR-148a-3p and miRNA-223-3p) had upregulated expression and 4 (miR-7d-5p, miR-210-3p, miR-324-5p and miR-654-3p) showed down-regulated expression. Target genes of these miRNAs were involved in bone development, cell proliferation in bone marrow, osteoblast development, negative regulation of osteoblast differentiation, and negative regulation of osteoclast development, as well as several osteogenesis related pathways. Canonical Wnt signaling pathway was selected for verification and function analysis. The expression of Wnt1, FZD10, LRP5, DVL2 and LEF1 was down-regulated significantly, while that of SFRP1, DKK1, and CHD8 was up-regulated markedly. In conclusion, these genes play important roles in OP, which improves our understanding of pathogenesis of OP.

Identification and differential expression of microRNAs in 1, 25-dihydroxyvitamin D3-induced osteogenic differentiation of human adipose-derived mesenchymal stem cells.

The aim of this study was to identify specific microRNAs (miRNAs) and their regulatory roles in the process of 1, 25-dihydroxyvitamin D3-induced (VD3-induced) osteogenic differentiation of human adipose-derived Mesenchymal stem cells (hAMSCs). The differentially expressed miRNAs in VD3-induced hAMSCs was examined. The putative target genes of these miRNAs were predicted. A total of 76 conserved miRNAs, including 18 miRNAs were significantly up-regulated and 58 miRNAs were significantly downregulated, and significantly differentially expressed between the two samples. The expression of 4 upregulated miRNAs (miR-1-3p, miR-1247-5p, miR-217, and miRNA-483) and 5 downregulated miRNAs (miR-1284, miR-218, miR-582-3p, miR-187-3p, and miRNA-122-5p) were verified. The highly enriched GOs and KEGG pathway showed target genes of these miRNAs were significantly involved in multiple biological processes (signal transduction, cell differentiation, cell adhesion and cell proliferation), and several osteogenic pathways (MAPK signaling pathway, TGF-ß/BMP signaling pathway, and Wnt signaling pathway). Finally, TGF-ß/BMP signaling pathway was selected for target verification and function analysis. We observed that a number of osteo-genes in the TGF-ß/BMP superfamily, such as BMPRI, BMPRII, TGFBRI, TGFBRII, BMP4, TGFß, Smad2, 3, 8, were predicted to be target gene of the differentially expressed miRNAs. Among them, TGFB, BMP4, BMPRI, and Smad8, which are positive regulators in osteoblast differentiation, were confirmed to be significantly up-regulated in VD3-induced cells by qRT-PCR; while Smad6 and activinRI, which are negative regulators of the TGF-ß/BMP superfamily, were shown to be significantly down-regulated. These results will help to understand the role of miRNA in the regulation of the osteogenic differentiation of hAMSCs.