The aim of the present study was to investigate the role of microRNA (miRNA or miR)-140 in C3H10T1/2 mesenchymal stem cells (MSCs). Cluster analysis was used to evaluate the miRNA expression profile. The expression level of miRNA‑140 was validated by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). TargetScan and microRNA.org databases were used to predict target miRNAs and cartilage‑associated target genes. Binding sites between miR‑140 and the target gene were predicted by bioinformatics software. A dual‑luciferase reporter assay was performed to determine whether miR‑140 could target C‑X‑C motif chemokine ligand 12 (CXCL12). Following the promotion/inhibition of miR‑140, 1, 7 and 14 days following transforming growth factor‑β3 (TGF‑β3)‑induction, western blotting was utilized to evaluate CXCL12 protein levels. MTT assays and alcian blue staining were applied to assess C3H10T1/2 MSC viability and chondrogenic differentiation, respectively. In the TGF‑β3‑induced group, RT‑qPCR verified that the mRNA level of Mus musculus (mmu)‑miR‑140 was significantly elevated when compared with the control group. miR‑140 was predicted to recognize and interact with CXCL12‑3'UTR and the dual luciferase reporter assay further validated that miR‑140 targeted the predicted region of CXCL12. CXCL12 was markedly decreased following miR‑140 overexpression and visibly increased following miR‑140 inhibition. In addition, the level of CXCL12 expression declined as the duration of induction increased. Following the promotion/inhibition of miR‑140, at 1 and 7 days following TGF‑β3‑induction, C3H10T1/2 MSCs inhibited or promoted cell viability, respectively, when compared with the control groups. In addition, in pellets achieved by chondrogenic differentiation following the induction of C3H10T1/2 MSCs for 7 days, alcian blue staining revealed no significant difference in characteristic extracellular matrix glycosaminoglycans between the miR‑140 up and downregulated groups, and their respective control groups. The present study concludes that miRNA‑140 inhibition promoted C3H10T1/2 MSC viability however, not C3H10T1/2 MSC differentiation by targeting and reducing CXCL12 protein levels during the process of TGF‑β3‑induced chondrogenic differentiation. In conclusion, the present study provided a potential target for the treatment of cartilage defection.
Abstract. The present study aimed to investigate the mechanisms underlying microRNA (miRNA)-mediated regulation of chondrogenic differentiation. Mouse embryo-derived stem cells C3H10T1/2 were cultured and chondrogenic differentiation was induced using transforming growth factor-β3 (TGF-β3). In addition, miRNA expression profiles were detected via miRNA array analysis, and quantitative polymerase chain reaction was performed to verify the differentially expressed miRNAs. Furthermore, bioinformatics software was used to predict the putative targets and the prediction was validated by dual-luciferase reporter assays and western blot analysis. In addition, cell proliferation and glycosaminoglycans were measured by a direct cell count method and alcian blue staining, respectively. Compared with the control group, 86 miRNAs were identified as differentially expressed in TGF-β3-induced cells and the expression levels of 28 miRNAs were increased while the remaining 58 miRNAs exhibited a decline in expression. Amongst the differentially expressed miRNAs, miR-30b expression was observed to have significantly decreased during chondrogenic differentiation. SOX9 is a target gene of miR-30b, and miR-30b inhibits SOX9 expression during chondrogenic differentiation. Furthermore, the alcian blue staining results demonstrated that miR-30b inhibited early chondrogenic differentiation. However, the data of the present study indicated that miR-30b had no influence on C3H10T1/2 cell line proliferation. In conclusion, miR-30b is a key negative regulator of TGF-β3-induced C3H10T1/2 cell chondrogenic differentiation, which functions by directly targeting SOX9.
Eukaryotic translation initiation factor 3H subunit (EIF3H) is a member of the EIF3 family and exhibits a central role in translation initiation in higher eukaryotes. Although EIF3H expression is upregulated in numerous tumour types, its potential role in human osteosarcoma (OS) has not yet been investigated. In the present study, it was demonstrated that EIF3H mRNA expression was upregulated in the human OS cell lines Saos-2 and U2OS. A recombinant lentivirus harbouring short hairpin RNA targeting EIF3H was constructed and successfully infected human OS Saos-2 and U2OS cells, resulting in 95% downregulated EIF3H expression compared with the respective control groups. Knockdown of EIF3H significantly inhibited the proliferation and colony formation of OS cells in vitro, and tumour growth in nude mice in vivo. Flow cytometry analysis revealed cell cycle arrest and promotion of apoptosis in OS cells with EIF3H knocked down. In conclusion, the results strongly suggested that EIF3H is a critical factor mediating the growth of OS cells and may represent a novel therapeutic target.
Background Bone is one of the most common sites of advanced tumors. However, there is currently a lack of population-based surveys for the incidence and prognosis of bone metastases in common solid cancers.Methods Patients with 12 types of primary cancer and bone metastases at initial diagnosis between 2010 and 2015 were identified using the Surveillance, Epidemiology, and End Results (SEER) database. The Kaplan-Meier method and Cox logistic regression were conducted to analyze survival and the effect of bone metastases on different cancers.Results We included 89,782 patients with bone metastases at cancer diagnosis. Lung cancer had the highest incidence (17.61%) of bone metastases at diagnosis in any stage, followed by liver cancer (6.29%), nasopharyngeal carcinoma (6.22%) and renal cancer (5.19%). Among patients with breast and prostate cancer, only 3.4% and 4.39%, respectively, were identified as having bone metastases at diagnosis. Breast cancer (32.1%), prostate cancer (25.2%), thyroid cancer (46.8%) and nasopharyngeal carcinoma (24.8%) patients with only bone metastasis have an over 20% five-year survival rate. Compared with patients at a stage previous to metastasis, bone metastasis significantly increased the risk of mortality and reduced survival time, especially for patients with prostate cancer (HR: 19.64, 95% CI 18.36 to 21.02). Concomitant other organ metastases make patient survival worse. Regarding the metastases of prostate cancer, bone metastases are the main type, while for colorectal cancer, bone metastases and concomitant visceral metastases mainly occur.Conclusions The findings of this study provide estimates of the incidence and prognosis of patients with bone metastases during the initial diagnosis of common solid cancers. In addition, we also clarified the degree to which bone metastasis affects patient survival. Patient prognosis depends on the primary type of cancer. These results can be used as a reference for the screening of metastases, and the optimization of personalized treatment options to improve the quality of life and survival of patients.
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