ATF1 regulates MAL2 expression through inhibition of miR-630 to mediate the EMT process that promotes cervical cancer cell development and metastasis.

BACKGROUND: The existence of activating transcription factor 1 (ATF1) could be employed as a clinical marker in the context of cervical cancer development, although its specific mechanism has not been fully clarified. METHODS: To evaluate the presence of ATF1, miR-630, and myelin and lymphocyte protein 2 (MAL2) in cervical malignancies, we conducted quantitative reverse transcription polymerase chain reaction, immunohistochemistry, and Western blot assays; further studied the expansion, migration, invasion and epithelial-mesenchymal transition (EMT) of cervical carcinoma cells using colony formation assay, transwell, loss cytometry, Western blot. Chromatin immunoprecipitation (ChIP) and RNA immunoprecipitation (RIP) were used to verify that ATF1 could directly transcriptionally repress miR-630; dual luciferase reporter assay and RIP assay were employed to confirm that miR-630 targeted to repress MAL2. RESULTS: In cervical cancer cases, elevated ATF1 expression and reduced miR-630 expression were detected, displaying a negative relationship between them. Inhibition of ATF1 hindered the growth, migration, infiltration, and EMT in cervical carcinoma cells, while upregulation of miR-630 mitigated the aggressive characteristics of these cells. ATF1 was found to transcriptionally repress miR-630 by TransmiR and ALGGEN prediction and ChIP validation. MicroRNA modulates gene expression and affects cancer progression, and we discovered that miR-630 regulates cancer progression by targeting and inhibiting MAL2. CONCLUSION: ATF1, which modulates the miR-630/MAL2 pathway, affects the EMT process and cervical carcinoma cell growth and spread. Therefore, ATF1 may serve as a promising marker and treatment target for cervical malignancies intervention.

Sclerostin Transduced Bone Marrow Mesenchymal Stem Cells Promote Fracture Healing in Rats Through the Wnt/ß-Catenin Signal Pathway.

The prognosis of fracture is directly related to several factors. Due to the limitations of existing treatment strategies, there are still many fractures with poor healing. Bone marrow mesenchymal stem cells (BMSCs) have the potential to differentiate into osteoblasts and chondrocytes. Therefore, BMSC transplantation is promised as an effective method for treating bone fractures. We aim to explore whether silently expressing sclerostin gene (SOST) can promote bone formation through the SOST/Wnt/ß-catenin signal pathway. We isolated rat BMSCs and the target gene (SOST shRNA) was transduced into them for osteogenic induction. The results showed that SOST significantly inhibited the proliferation and osteogenic differentiation of BMSCs during osteogenic induction, whereas silently expressing SOST not only increased the number of surviving BMSCs but also promoted the expression of osteogenesis-related proteins RUNX2, osteoprotegerin, Collagen I (COL-I), and bone morphogenetic protein-2 during osteogenic induction. The results of imaging examination in rats show that downregulating the expression of SOST can promote the formation of bony callus and the transformation of cartilage tissue into normal bone tissue, and then accelerate the healing of osteoporotic fracture. In addition, we also found that SOST silencing can activate the Wnt/ß-catenin pathway to achieve these effects. In conclusion, SOST silencing can promote the proliferation and osteogenic differentiation of BMSCs in situ, and therefore may enhance the therapeutic efficiency of BMSC transplantation in OPF.

Icariin inhibits osteoblast ferroptosis via Nrf2/HO-1 signaling and enhances healing of osteoporotic fractures.

The incidence of osteoporotic fractures is increasing every year because of population aging around the world. The reduced osteoblast activity in osteoporotic fracture has been associated with ferroptosis. A recent study showed that the antioxidant icariin (ICA) reduced iron deposition in the bone marrow of osteoporotic mice, although the underlying regulatory mechanisms were not explored. The objective of present study was to assess the therapeutic effects of ICA in a rat osteoporotic fracture model, with particular focus on its impact on ferroptosis. Primary rat osteoblasts were exposed to the ferroptosis inducer erastin, and then treated with ICA or the ferroptosis inhibitor ferrostatin-1 (Fer-1) as the positive control group. The levels of Nrf2 signaling factors and osteogenesis-related factors were examined by RT-PCR and western blotting. An osteoporotic fracture model was established in rats, and the effect of ICA on bone formation was evaluated by X-ray, Micro CT analysis, histological examination and Safranin O staining. Furthermore, the levels of GPX4, Bax, Nrf2 and Runx2 proteins at the fracture site were examined by immunohistochemistry. ICA significantly reduced ROS levels in the erastin-treated osteoblasts, and downregulated glutathione peroxidase 4 (GPX4) and cystine glutamate antiporter (SLC7A11). Moreover, ICA also upregulated Nrf2, NQO-1, HO-1, Runx2, ALP, OPG and OCN in these cells, which was reversed by inhibitors of the Nrf2 signaling pathway and Nrf2 silencing. X-ray and Micro CT analysis showed that ICA increased the trabecular bone and promoted callus formation in the osteoporotic fracture model, and also enhanced the transition from fibrous to osseous callus. Furthermore, ICA upregulated GPX4, Nrf2 and Runx2 at the fracture site, and significantly reduced the expression of the apoptotic genes of Bax. Taken together, our findings indicate that ICA promotes osteoporotic fracture healing by inhibiting osteoblast ferroptosis via activation of the antioxidant Nrf2/HO-1 signaling pathway.

[Corrigendum] Icaritin promotes the osteogenesis of bone marrow mesenchymal stem cells via the regulation of sclerostin expression.

Following the publication of the above article, the authors have contacted the Editorial Office to explain that they had assembled the cellular morphological images in Fig. 1A on p. 819 incorrectly; essentially, the cell morphology of 2 passages of hBMSCs (centre panel) should have been shown as the data panel for 3 passages of hBMSCs (right-hand panel), and likewise, the cell morphology of 3 passages of hBMSCs should have been shown as the data panel for 2 passages of hBMSCs. The revised version of Fig. 1 is shown below. The authors confirm that the errors associated with this figure did not have any significant impact on either the results or the conclusions reported in this study, and are grateful to the Editor of International Journal of Molecular Medicine for allowing them the opportunity to publish this Corrigendum. Furthermore, they apologize to the readership of the Journal for any inconvenience caused. [International Journal of Molecular Medicine 45: 816-824, 2020; DOI: 10.3892/ijmm.2020.4470].

Curcumin Inhibits the Growth and Metastasis of Melanoma via miR-222-3p/SOX10/Notch Axis.

Background: The aim of this study was to investigate the effect of curcumin on melanoma and its mechanism. Methods: Curcumin (0, 0.125, 0.25, or 0.5 mg/ml) was utilized to treat A375 and HT144 cell lines. The MTT analysis was used to confirm the proliferation ability. Wound healing and transwell analysis showed the migration and invasion ability. Immunofluorescence assay was used to demonstrate the effect of curcumin on SOX10 expression. Multiple bioinformatic analysis to confirm the SOX10 associated miRNA. The correlation of miR-222-3p and SOX10 was detected by Luciferase reporter assays. qRT-PCR showed the miR-222-3p level. Western blot analyzed the expression of SOX10, Notch1, and HES1 in melanoma cell treated with or without miR-222-3p inhibitor. Results: Curcumin could inhibit the proliferation, migration, and invasion of melanoma cells. Furthermore, curcumin repress the expression of SOX10, Notch1, and HES-1, and increase the expression of miR-222-3p. And the miR-222-3p could directly target to SOX10 mRNA to inhibit its expression. In addition, inhibition of miR-222-3p expression reversed the inhibitory effect of curcumin the growth of melanoma cells. Conclusion: Curcumin enhances the miR-222-3p level to reduce SOX10 expression, and ultimately inactivates the Notch pathway in repressing melanoma proliferation, migration, and invasion.

Terahertz-Based Method for Accurate Characterization of Early Water Absorption Properties of Epoxy Resins and Rapid Detection of Water Absorption.

Moisture is detrimental to the performance of epoxy resin material for electrical equipment in long-term operation and insulation. Therefore, moisture absorption is one of the critical indicators for insulation of the material. However, some relevant test methods, e.g., the direct weighing method, are time-consuming, and it usually takes months to complete a test. For this, it is necessary to have some modification to save the test time. Firstly, the study analyzes the present prediction method (according to ISO 62:2008). Under the same accuracy, the time required is reduced from 104 days to 71 days. Subsequently, the Langmuir curve-fitting method for water absorption of epoxy resin is analyzed, and the initial values of diffusion coefficient, bonding coefficient, and de-bonding coefficient are determined based on the results of molecular simulation, relevant experiment, and literature review. With the optimized prediction model, it takes only 1.5 days (reduced by 98% as compared with the standard prediction method) to determine the moisture absorbability. Then, the factors influencing the prediction accuracy are discussed. The results have shown that the fluctuation of balance at the initial stage will affect the test precision significantly. Accordingly, this study proposes a quantitative characterization method for initial trace moisture based on the terahertz method, by which the trace moisture in epoxy resin is represented precisely through the established terahertz time-domain spectroscopy system. When this method is used to predict the moisture absorbability, the experimental time may be further shortened by 33% to 1 day. For the whole water absorption cycle curve, the error is less than 5%.

SOX10 Knockdown Inhibits Melanoma Cell Proliferation via Notch Signaling Pathway.

PURPOSE: Melanoma is a serious and malignant disease worldwide. Seeking diagnostic markers and potential therapeutic targets is urgent for melanoma treatment. SOX10, a member of the SoxE family of genes, is a transcription factor which can regulate the transcription of a wide variety of genes in multiple cellular processes. METHODS: The mRNA level and protein expression of SOX10 is confirmed by bioinformatic analysis and IHC staining. MTT, clone formation and EdU analysis showed that SOX10 knockdown (KD) could significantly inhibit melanoma cell proliferation. FACS analysis showed that SOX10 KD could markedly enhance the level of cell apoptosis. The downstream target signaling pathway is predicted by RNA-seq based on the public GEO database. The activation of Notch signaling mediated by SOX10 is tested by qPCR and Western blot. RESULTS: Ectopic upregulation of SOX10 was found in melanoma patient tissues compared to normal nevus tissues in mRNA and protein levels. Furthermore, both mRNA and protein level of SOX10 were negatively correlated with melanoma patient's prognosis. SOX10 knockdown could obviously suppress the proliferation ability of melanoma cells by inactivating Notch signaling pathway. CONCLUSION: Our study confirmed that SOX10 is an oncogene and activate Notch signaling pathway, which suggests the potential treatment for melanoma patients by target SOX10/Notch axis.

Acid-Etched Co3O4 Nanoparticles on Nickel Foam: The Highly Reactive (311) Facet and Enriched Defects for Boosting Methanol Oxidation Electrocatalysis.

The confirmation and regulation of active sites are particularly critical for the design of methanol oxidation reaction (MOR) catalysts. Here, an acid etching method for facet control combined with defect construction was utilized to synthesize Co3O4 nanoparticles on nickel foam for preferentially exposing the (311) facet with enriched oxygen vacancies (VO). The acid-leached oxides exhibited superior MOR activity with a mass activity of 710.94 mA mg-1 and an area-specific activity of 3.390 mA cm-2 as a result of (i) abundant active sites for MOR promoted by VO along with the highly active (311) facet being exposed and (ii) phase purification-reduced adsorption energy (Eads) of methanol molecules. Ex situ X-ray photoelectron spectroscopy proved that highly active CoOOH obtained via the activation of plentiful Co2+ effectively improved the MOR. Density functional theory calculations confirmed that the selective exposed (311) facet has the lowest Eads for CH3OH molecules. This work puts forward acid etching as the facet modification and defect engineer for nanostructured non-noble catalysts, which is expected to result in superior electrochemical performance required for advanced alkaline direct methanol fuel cells.

Rutin maintains redox balance to relieve oxidative stress induced by TBHP in nucleus pulposus cells.

Rutin is well known for its anti-inflammatory and antioxidant properties against oxidative stress. However, its protective function in nucleus pulposus cells (NPCs) remains unclear. This study was aimed to explore the effects of rutin on oxidative stress in NPCs. Primary NPCs were obtained from 1-mo-old SD rats. The NPCs were treated with tert-butyl hydrogen peroxide (TBHP) to obtain the oxidative stress, and different concentrations of rutin were used to observe its influence on the oxidative stress in NPCs. Fluorescent probe DCFH-DA was used to detect reactive oxide species (ROS). The antioxidant proteins and genes of heat shock protein 70 (HSP70), manganese superoxide dismutase (Mn-SOD), catalase, aggrecan and collagen II in NPCs were measured by western blot and real-time PCR. With the stimulation of TBHP, the content of ROS in NPCs increased significantly and showed solubility correlation. Rutin effectively reduced the accumulation of ROS in a dose-dependent manner. The antioxidant proteins of HSP70, Mn-SOD, and catalase and the matrix proteins of aggrecan and collagen II decreased remarkably with the stimulation of TBHP, while the matrix metalloproteinase-13 (MMP-13) significantly increased after TBHP intervention. Rutin boosted the expressions of the HSP70, Mn-SOD, and catalase, elevated the contents of aggrecan and collagen II, and inhibited the expression of MMP-13 in NPCs. The findings of this study suggested that rutin is able to reverse oxidative stress and maintain cellular function of NPCs, and it was indicated that rutin could be a possible therapeutic option for intervertebral disc degeneration.

Icariin Promotes the Osteogenesis of Bone Marrow Mesenchymal Stem Cells through Regulating Sclerostin and Activating the Wnt/ß-Catenin Signaling Pathway.

Osteoporosis (OP) is a metabolic disease characterized by decreased bone mass and increased risk of fragility fractures, which significantly reduces the quality of life. Stem cell-based therapies, especially using bone marrow mesenchymal stem cells (BMSCs), are a promising strategy for treating OP. Nevertheless, the survival and differentiation rates of the transplanted BMSCs are low, which limits their therapeutic efficiency. Icariin (ICA) is a traditional Chinese medicine formulation that is prescribed for tonifying the kidneys. It also promotes the proliferation and osteogenic differentiation of BMSCs, although the specific mechanism remains unclear. Based on our previous research, we hypothesized that ICA promotes bone formation via the sclerostin/Wnt/ß-catenin signaling pathway. We isolated rat BMSCs and transfected them with sclerostin gene (SOST) overexpressing or knockdown constructs and assessed osteogenic induction in the presence or absence of ICA. Sclerostin significantly inhibited BMSC proliferation and osteogenic differentiation, whereas the presence of ICA not only increased the number of viable BMSCs but also enhanced ALP activity and formation of calcium nodules during osteogenic induction. In addition, the osteogenic genes including Runx2, ß-catenin, and c-myc as well as antioxidant factors (Prdx1, Cata, and Nqo1) were downregulated by sclerostin and restored by ICA treatment. Mechanistically, ICA exerted these effects by activating the Wnt/ß-catenin pathway. In conclusion, ICA can promote the proliferation and osteogenic differentiation of BMSCs in situ and therefore may enhance the therapeutic efficiency of BMSC transplantation in OP.

Template-Directing Coupled with Chemical Activation Methodology-Derived Hexagon-like Porous Carbon Electrode with Outstanding Compatibility to Electrolytes and Low-Temperature Performance.

The conversion of asphalt into hexagon-like porous carbon (HPC) with a micro-mesoporous structure is realized by the coupling of template-directing and chemical activation methodologies. The specific surface area of HPC can reach up to 1356 m2 g-1 even at such a low-proportioned dosage of activator (0.5-fold) and is also larger than those of template-directed carbon and activation-derived carbon, as it benefited from the coupling merits of template-directing and chemical activation. Excellent capacitive-energy-storage behavior with respect to rate capability, capacitance retention, and durability are delivered by HPC//HPC symmetric supercapacitors assembled with aqueous and organic electrolytes. This great compatibility for different kinds of electrolytes and electrode properties is owed to the robust hexagon-like microarchitecture feature associated with hierarchical pore structure, which not only hinders the stacking between each other but also provides a buffer function for the volume variation and sufficient active sites for the storage of electrolyte ions. The drastic temperature variation has almost no influence on the diffusion and transfer rate of electrolyte ions, further evidencing the advanced feature of the hierarchical pore structure. Additionally, HPC//Li4Ti5O12 LIC assembled with the Li-based electrolyte also presents a superior Ragone performance. The coexistence of micro- and mesopores for the HPC makes it an attractive electrode material for various capacitive-energy-storage devices. This work provides a promising way to realize the plasticity of pore channels and mass production of high capacitive storage ability of electrode material via the combination of template-directing and chemical activation strategies.

Heterostructure Ni3S4-MoS2 with interfacial electron redistribution used for enhancing hydrogen evolution.

Developing highly effective and inexpensive electrocatalysts for hydrogen evolution reaction (HER), particularly in a water-alkaline electrolyzer, are crucial to large-scale industrialization. The earth-abundant molybdenum disulfide (MoS2) is an ideal electrocatalyst in acidic media but suffers from a high overpotential in alkaline solution. Herein, nanospherical heterostructure Ni3S4-MoS2 was obtained via a one-pot synthesis method, in which Ni3S4 was uniformly integrated with MoS2 ultrathin nanosheets. There were abundant heterojunctions in the as-synthesized catalyst, which were verified by X-ray photoelectron spectroscopy (XPS) and high-resolution transmission electron microscopy (HRTEM). The structure features with interfacial electron redistribution was proved by XPS and density functional theory (DFT) calculations, which offered several advantages to promote the HER activity of MoS2, including increased specific surface area, exposed abundant active edge sites and improved electron transfer. Ni3S4-MoS2 exhibited a low overpotential of 116 mV at 10 mA cm-2 in an alkaline solution with a corresponding Tafel slope of 81 mV dec-1 and long-term stability of over 20 h. DFT simulations indicated that the synergistic effects in the system with the chemisorption of H on the (002) plane of MoS2 and OH on the (311) plane of Ni3S4 accelerated the rate-determining water dissociation steps of HER. This study provides a valuable route for the design and synthesis of inexpensive and efficient HER electrocatalyst, heterostructure Ni3S4-MoS2.

MEG3 Induces Cervical Carcinoma Cells' Apoptosis Through Endoplasmic Reticulum Stress by miR-7-5p/STC1 Axis.

Background: Many patients with advanced cervical cancer (CC) have a poor prognosis and their mortality rank the first among women with malignant tumors. It's essential to explore the molecular mechanism of CC in clinical practice. Long noncoding RNA maternally expressed gene 3 (MEG3) has been reported to downregulate in CC tissues. However, the underlying mechanism of MEG3 in CC remains poorly elaborated. The current study aimed to explore the potential mechanism of MEG3 inducing endoplasmic reticulum stress (ERs)-mediated apoptosis of CC cells. Methods: The expression of MEG3 and miR-7-5p in CC tissues and cell lines was verified by quantitative reverse transcription/polymerase chain reaction (qRT-PCR). The vector of MEG3, miR-7-5p inhibitor, and sh-SCT1 were transfected into CC cell lines, and their expression was tested by qRT-PCR. Flow cytometry was used to detect apoptosis, and ERs-related protein expression was performed by Western blot. The regulatory relationship between MEG3/SCT1 and miR-7-5p was validated by Dual luciferase reporter assay. Results: CC tissues and cell lines showed downregulated MEG3 and STC1, and upregulated miR-7-5p. Overexpression of MEG3 or miR-7-5p inhibition induced ERs-triggered apoptosis of CC cells. In addition, sh-STC1 can reverse the effects of overexpressing MEG3 on CC cell apoptosis. In addition, dual luciferase reporter assay revealed that miR-7-5p can directly target to MEG3 and STC1. Conclusion: MEG3, act as a competing endogenous RNA of miR-7-5p, accelerates ERs-mediated apoptosis of CC cells through regulating SCT1 expression.

Trichostatin A Induces Autophagy in Cervical Cancer Cells by Regulating the PRMT5-STC1-TRPV6-JNK Pathway.

OBJECTIVE: The aim of this study was to investigate the effects of trichostatin A (TSA) on cervical cancer and the related mechanisms. METHODS: The HeLa and Caski cervical cancer cell lines were treated with different concentrations of TSA. Cell viability was measured by MTT assays. Cell apoptosis was analysed using flow cytometry. Expression of transient receptor potential cation channel, subfamily V, member 6 (TRPV6), protein arginine methyltransferase 5 (PRMT5), and stanniocalcin 1 (STC1) was determined by qRT-PCR and Western blotting. Protein levels of LC3 II/I, beclin1, p62, JNK, and p-JNK were detected by Western blotting. RESULTS: Treatment with TSA significantly decreased HeLa and Caski cell viability and enhanced the apoptosis rate in a dose-dependent manner. TSA markedly elevated beclin1 protein levels and the LC3 II/I ratio and significantly reduced p62 levels in a dose-dependent manner. In addition, TSA (1 µM) significantly suppressed PRMT5 and TRPV6 levels and enhanced STC1 and p-JNK levels. The lysosomal inhibitor bafilomycin-A1 synergistically enhanced the TSA-mediated increase in autophagic flux. Either the overexpression of TRPV6 or the inhibition of JNK signalling markedly enhanced cell viability, inhibited apoptosis, and autophagy and reduced p-JNK levels in TSA-treated cells. The inhibition of STC1 significantly increased TRPV6 protein levels and reduced p-JNK levels. Overexpression of PRMT5 dramatically decreased STC1 and p-JNK protein levels and increased TRPV6 levels. CONCLUSION: TSA suppresses cervical cancer cell proliferation and induces apoptosis and autophagy through regulation of the PRMT5/STC1/TRPV6/JNK axis.

The Hyperglycemia and Hyperketonemia Impaired Bone Microstructures: A Pilot Study in Rats.

Though diabetes mellitus (DM) is one of the known causes of osteoporosis, it is also realized that ketogenic diet (KD), an effective regimen for epilepsy, impairs bone microstructures. However, the similarities and differences of effects between these two factors are still unknown. The purpose of this study is to identify different effects between hyperglycemia and hyperketonemia, which are manifestations of DM and KD, on bone in rats. Thirty male Sprague-Dawley rats were randomly divided into three groups: the sham, DM, and KD groups. Hyperglycemia was achieved by intravenous injection of streptozotocin in DM group, while hyperketonemia was induced by application of ketogenic diet (carbohydrates-to-fat as 1:3) in KD group. The body weight, blood ketone body, and blood glucose were recorded, and the bone turnover markers, bone length, bone microstructures, bone biomechanics and histomorphology were measured after 12 weeks intervention. Compared with the control and KD groups, a significant body weight loss was found in the DM group, and the bone lengths of tibia and femur of the group were shortened. The blood glucose and blood ketone were noticeably increased in the DM and KD rats, respectively. Microstructures and properties of cancellous bone were significantly deteriorated in both the DM and KD groups compared with the sham group, as the bone volumes were decreased and the bone trabecula structures were disturbed. Meanwhile, the thickness and strength of cortical bone was reduced more in the DM group than those in the sham and KD groups. The HE staining showed that bone trabecula was significantly decreased in both the DM and KD groups, and more adipose tissue was observed in the KD rats. The activity of osteoblasts was decreased more in both the KD and DM groups than that in the sham group, while the activity of osteoclasts of the two groups was remarkably increased. The present study indicates that both hyperglycemia and hyperketonemia have adverse effects on bone. Therefore, it is worth paying more attention to the bone status of patients with hyperglycemia and hyperketonemia in clinic.

miR-19 Promotes Cell Proliferation, Invasion, Migration, and EMT by Inhibiting SPRED2-mediated Autophagy in Osteosarcoma Cells.

Osteosarcoma is an aggressive malignancy with rapid development and poor prognosis. microRNA-19 (miR-19) plays an important role in several biological processes. Sprouty-related EVH1 domain protein 2 (SPRED2) is a suppressor of extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) signaling to inhibit tumor development and progression by promoting autophagy. In this study, we investigated the roles of miR-19, SPRED2, and autophagy in osteosarcoma. We detected the expression of miR-19, SPRED2, epithelial-mesenchymal transition (EMT) markers, and autophagy-related proteins via quantitative real-time polymerase chain reaction or western blot. To evaluate the function of miR-19 and SPRED2, we used MTT and colony formation assays to detect cell proliferation, Transwell, and wound-healing assays to detect cell invasion and migration. Targetscan and luciferase reporter assays confirmed the relationship between SPRED2 and miR-19. The expression of miR-19 was significantly upregulated in osteosarcoma, while SPRED2 was downregulated. miR-19 inhibitor reduced cell proliferation, invasion, migration, and EMT, while its cell biological effects were partially reversed by addition of autophagy inhibitor 3-methyladenine (3-MA) or SPRED2 siRNA in osteosarcoma. SPRED2, a suppressor of ERK/MAPK pathway that is known to trigger autophagy, was identified as a direct target of miR-19. SPRED2 overexpression increased cell proliferation, invasion, migration, and EMT by promoting autophagy, and the effects could be inhibited by 3-MA. Collectively, these findings reveal an underlying mechanism for development of osteosarcoma. miR-19 was upregulated in osteosarcoma cells, and negatively regulated SPRED2, thus promoting the malignant transformation of osteosarcoma cells via inhibiting SPRED2-induced autophagy. Therefore, miR-19/SPRED2 may be a potential target for the treatment of osteosarcoma.

CircSAMD4A regulates cell progression and epithelial­mesenchymal transition by sponging miR­342­3p via the regulation of FZD7 expression in osteosarcoma.

Osteosarcoma (OS) is a primary malignant tumor with a complex etiology. Therefore, research into the pathogenesis of osteosarcoma is considered a priority. Circular RNAs play important roles in cell metabolism and in the immune response and are closely associated with cancer treatment. However, research into the association of circular RNAs with osteosarcoma is limited. In the present study, CircSAMD4A was validated by RT­qPCR and agarose gel electrophoresis. CircSAMD4A and miR­342­3p expression was detected by RT­qPCR. The relative protein expression levels were measured by western blot analysis. MTT assay and flow cytometry were used to detect cell cytotoxicity and apoptosis, respectively. Transwell assay was applied to assess cell migration and invasion. Dual­luciferase reporter assay was used to determine the association among CircSAMD4A, Frizzled­7 (FZD7) and miR­342­3p. In vivo, subcutaneous tumor formation assay was performed in an experiment with nude mice. The results revealed that the expression levels of CircSAMD4A and FZD7 were upregulated, while those of miR­342­3p were downregulated in OS tissues and cells. The inhibition of CircSAMD4A suppressed cell progression and epithelial­mesenchymal transition (EMT), and promoted cell apoptosis in OS. The reduction of miR­342­3p reversed the effects of CircSAMD4A downregulation on cell cytotoxicity, migration, invasion, apoptosis and EMT in OS, while FZD7 overexpression blocked the effect of miR­342­3p upregulation on OS progression. The suppressive effect of sh­CircSAMD4A on tumor growth was thus verified in OS. Overall, the present study demonstrated that CircSAMD4A affected cell cytotoxicity, invasion, apoptosis, migration and EMT by regulating the miR­342­3p/FDZ7 axis in OS, thereby providing a novel regulatory mechanism and a potential therapeutic target for OS.

Icaritin promotes the osteogenesis of bone marrow mesenchymal stem cells via the regulation of sclerostin expression.

Icaritin, a metabolite of icariin, is a potent promoter of bone marrow­derived mesenchymal stem cells (BMSCs) osteogenesis, but the underlying mechanisms remain unclear. To examine the effects of icaritin on osteogenic differentiation, BMSCs were exposed to osteogenic induction medium with or without icaritin pretreatment in the present study. It was identified that icaritin (0.01­1 µM) exhibited no cytotoxicity on the proliferative abilities of the BMSCs. Icaritin at 1 µM increased alkaline phosphatase activity, mineral deposition and osteoblast­specific gene expression. Treatment with 1 µM Icaritin upregulated osteocalcin, RUNX family transcription factor 2, tissue­nonspecific alkaline phosphatase and ß­catenin, and suppressed sclerostin (SOST) gene expression in different stages of osteogenic differentiation. It was also demonstrated that SOST overexpression inhibited icaritin­induced osteogenesis. The western blot analysis data suggested that ICI 182780, which causes estrogen receptor α (ERα) degradation, reversed the icaritin­induced decrease in SOST expression, which was inconsistent with the results of immunofluorescence analysis. In conclusion, icaritin was demonstrated to promote the osteogenesis of hBMSCs by downregulating SOST expression, and icaritin­induced suppression of SOST was regulated in part via the Wnt/ß­catenin/ERα axis.

The Ginsenoside Exhibits Antiosteoporosis Effects in Ketogenic-Diet-Induced Osteoporosis via Rebalancing Bone Turnover.

Ginsenoside is widely used in China for therapeutic and healthcare practice. Ginsenoside-Rb2 shows the antiosteoporosis effects in ovariectomized rodents. However, the protective effects on osteoporosis induced by ketogenic diet (KD) remain unknown. Therefore, this study aimed at evaluating the effects of ginsenoside-Rb2 on KD-induced osteoporosis. Thirty mice were randomly divided into three groups: sham, KD, and KD + Rb2. Bone microstructures, biomechanical properties, concentrations of serum bone alkaline phosphatase (BALP) and tartrate-resistant acid phosphatase (TRACP), and protein expression of osteocalcin (OCN), peroxisome proliferation-activated receptor γ (PPAR-γ), cathepsin K, and TRAP were evaluated after a 12-week intervention. The results show that KD induced significant bone loss and biomechanical impairment. Ginsenoside-Rb2 attenuated significant bone loss and maintained biomechanics in cancellous bone. The bone volume fraction increased from 2.3 to 6.0% in the KD + Rb2 group than that in the KD group. Meanwhile, ginsenoside-Rb2 effectively maintained biomechanical strengths in cancellous bone, increased serum BALP and decreased TRACP, and upregulated OCN and downregulated TRAP, PPAR-γ, and cathepsin K in the KD mice. This study demonstrated that ginsenoside-Rb2 retards bone loss and maintains biomechanics with KD. The underlying mechanism might be that ginsenoside-Rb2 inhibits bone resorption process and induces osteogenic differentiation, providing evidence for ginsenoside as being an alternative option for osteoporosis induced by KD.

Polydatin promotes the osteogenic differentiation of human bone mesenchymal stem cells by activating the BMP2-Wnt/ß-catenin signaling pathway.

Steroid-induced osteonecrosis of the femoral head (SONFH) is a refractory disease induced by glucocorticoids. Marrow mesenchymal stem cells (MSCs) differentiate into multiple bone matrix cells and have been used as cell-based therapies to treat ONFH. However, the osteogenesis of MSCs isolated from patients with SONFH is significantly decreased. Polydatin has been widely used in traditional Chinese remedies due to its multiple pharmacological actions. As shown in our previous study, Polydatin protects from oxidative stress and promotes BMSC migration. However, little is known about its role in BMSC (Bone marrow mesenchymal stem cells) osteogenesis; therefore, we further investigated the effect and mechanism of Polydatin in hBMSC osteogenesis. The ability of Polydatin to promote the proliferation and osteogenic differentiation of hBMSCs was determined using the MTT assay, ALP staining and the ALP activity assay. Next, qPCR and western blotting were performed to measure the levels of genes and proteins related to the osteogenesis of hBMSCs. Then, the effect of Polydatin on the nuclear translocation of ß-catenin was determined using immunofluorescence staining. Polydatin (30 µM) markedly enhanced the proliferation of hBMSCs and alkaline phosphatase (ALP) activity. Additionally, it also significantly upregulated the expression of osteogenic genes (Runx2, osteopontin, DLX5, osteocalcin, collagen type I and BMP2) and components of the Wnt signaling pathway (ß-catenin, Lef1, TCF7, c-jun, c-myc and cyclin D). These osteogenesis-potentiating effects of Polydatin were blocked by Noggin, an inhibitor of the BMP pathway, and DKK1, an inhibitor of the Wnt/ß-catenin pathway. However, DKK1 did not affect Polydatin-induced BMP2 expression. Based on our results, Polydatin promotes the proliferation and osteogenic differentiation of hBMSCs through the BMP2-Wnt/ß-catenin signaling pathway.