Identification of key genes associated with cervical cancer based on bioinformatics analysis.

BACKGROUND: Cervical cancer has extremely high morbidity and mortality, and its pathogenesis is still in the exploratory stage. This study aimed to screen and identify differentially expressed genes (DEGs) related to cervical cancer through bioinformatics analysis. METHODS: GSE63514 and GSE67522 were selected from the GEO database to screen DEGs. Then GO and KEGG analysis were performed on DEGs. PPI network of DEGs was constructed through STRING website, and the hub genes were found through 12 algorithms of Cytoscape software. Meanwhile, GSE30656 was selected from the GEO database to screen DEMs. Target genes of DEMs were screened through TagetScan, miRTarBase and miRDB. Next, the hub genes screened from DEGs were merged with the target genes screened from DEMs. Finally, ROC curve and nomogram analysis were performed to assess the predictive capabilities of the hub genes. The expression of these hub genes were verified through TCGA, GEPIA, qRT-PCR, and immunohistochemistry. RESULTS: Six hub genes, TOP2A, AURKA, CCNA2, IVL, KRT1, and IGFBP5, were mined through the protein-protein interaction network. The expression of these hub genes were verified through TCGA, GEPIA, qRT-PCR, and immunohistochemistry, and it was found that TOP2A, AURKA as well as CCNA2 were overexpressed and IGFBP5 was low expression in cervical cancer. CONCLUSIONS: This study showed that TOP2A, AURKA, CCNA2 and IGFBP5 screened through bioinformatics analysis were significantly differentially expressed in cervical cancer samples compared with normal samples, which might be biomarkers of cervical cancer.

Frizzled-7-targeting antibody (SHH002-hu1) potently suppresses non-small-cell lung cancer via Wnt/ß-catenin signaling.

Non-small-cell lung cancer (NSCLC) is one of the deadliest cancers worldwide, and metastasis is considered one of the leading causes of treatment failure in NSCLC. Wnt/ß-catenin signaling is crucially involved in epithelial-mesenchymal transition (EMT), a crucial factor in promoting metastasis, and also contributes to resistance developed by NSCLC to targeted agents. Frizzled-7 (Fzd7), a critical receptor of Wnt/ß-catenin signaling, is aberrantly expressed in NSCLC and has been confirmed to be positively correlated with poor clinical outcomes. SHH002-hu1, a humanized antibody targeting Fzd7, was previously successfully generated by our group. Here, we studied the anti-tumor effects of SHH002-hu1 against NSCLC and revealed the underlying mechanism. First, immunofluorescence (IF) and near-infrared (NIR) imaging assays showed that SHH002-hu1 specifically binds Fzd7+ NSCLC cells and targets NSCLC tissues. Wound healing and transwell invasion assays indicated that SHH002-hu1 significantly inhibits the migration and invasion of NSCLC cells. Subsequently, TOP-FLASH/FOP-FLASH luciferase reporter, IF, and western blot assays validated that SHH002-hu1 effectively suppresses the activation of Wnt/ß-catenin signaling, and further attenuates the EMT of NSCLC cells. Finally, the subcutaneous xenotransplanted tumor model of A549/H1975, as well as the popliteal lymph node (LN) metastasis model, was established, and SHH002-hu1 was demonstrated to inhibit the growth of NSCLC xenografts and suppress LN metastasis of NSCLC. Above all, SHH002-hu1 with selectivity toward Fzd7+ NSCLC and the potential of inhibiting invasion and metastasis of NSCLC via disrupting Wnt/ß-catenin signaling, is indicated as a good candidate for the targeted therapy of NSCLC.

Rapid thermal evaporation for cadmium selenide thin-film solar cells.

Cadmium selenide (CdSe) belongs to the binary II-VI group semiconductor with a direct bandgap of ∼1.7 eV. The suitable bandgap, high stability, and low manufacturing cost make CdSe an extraordinary candidate as the top cell material in silicon-based tandem solar cells. However, only a few studies have focused on CdSe thin-film solar cells in the past decades. With the advantages of a high deposition rate (∼2 °m/min) and high uniformity, rapid thermal evaporation (RTE) was used to maximize the use efficiency of CdSe source material. A stable and pure hexagonal phase CdSe thin film with a large grain size was achieved. The CdSe film demonstrated a 1.72 eV bandgap, narrow photoluminescence peak, and fast photoresponse. With the optimal device structure and film thickness, we finally achieved a preliminary efficiency of 1.88% for CdSe thin-film solar cells, suggesting the applicability of CdSe thin-film solar cells.

Short-term clinical effect of 3D printing techniques on the correction of complex malformations.

BACKGROUND: To explore the clinical effects of 3D printing techniques on the correction of complex malformation. METHOD: A computed tomography (CT) scan was used to collect data on malformations of patients and the orthopedic plan was made by virtual manipulation of the reality before surgery. The results of the virtual orthopedics were compared with the expected results. A guide plate for osteotomy was also utilized when necessary. The actual operation was carried out according to the plan. RESULTS: The average age of the 11 patients was 19.09 years (19.09 ± 6.93 years) and the average follow-up was 16 months (16 ± 15.11 months). The symptoms were obviously improved. The preoperative World Health Organization Disability Assessment Schedule (WHODAS 2.0) score, modified Barthel index and Functional Independence Measure (FIM) score in patients were 70.45 ± 15.75, 96.55 ± 3.78 and 121.36 ± 4.15, respectively and correspondingly 53 ± 12.75, 98.82 ± 1.66 and 123.82 ± 4.60 after surgery, respectively. There were significant differences before and after surgery (P < 0.05). CONCLUSION: The use of 3D printing technology can provide intuitive and accurate help for the correction of complex limb malformations and greatly facilitates the communication between doctors and patients. The FIM score is suitable for the evaluation of the curative effect before and after the treatment of patients with complex malformations.

Androgen Receptor Is Inactivated and Degraded in Bladder Cancer Cells by Phenyl Glucosamine via miR-449a Restoration.

BACKGROUND Bladder cancer caused by exposure to aniline dyes, chronic cystitis, and smoking is detected in approximately 70 000 new cases annually. In the USA alone, it leads to 15 000 deaths every year. In the present study, we investigated the role of 3-((4'-amino-[1,1'-biphenyl]-4-yl)amino)-4-bromo-5-oxo-2,5-dihydrofuran-2-yl acetate (ABDHFA) in the inhibition of bladder cancer cell viability. MATERIAL AND METHODS Viability of cells was examined using MTT assay and distribution of cell cycle was assessed by flow cytometry. Expression of cyclin D1, androgen, prostate-specific antigen (PSA), and miR-449a was analyzed using Western blot and quantitative real-time polymerase chain reaction assays. RESULTS The results demonstrated that ABDHFA treatment inhibited viability of UMUC3 and TCCSUP AR-positive bladder cancer cells. ABDHFA treatment led to break-down of AR in UMUC3 and TCCSUP cells after 48 h in a dose-dependent manner. Up-regulation of miR-449a by lentivirus transfection down-regulated the AR signalling pathway. In UMUC3 and TCCSUP cells, ABDHFA treatment led to inhibition of mRNA and protein expression corresponding to AR. CONCLUSIONS In summary, the present study demonstrates that proliferation of AR-positive bladder carcinoma cells is markedly reduced by ABDHFA treatment through arrest of cell cycle and degradation of AR protein. Thus, ABDHFA, a novel compound, can be used for the treatment of bladder cancer.

Role of RANK and Akt1 activation in human osteosarcoma progression: A clinicopathological study.

The receptor activator of nuclear factor κB (RANK) axis is the fundamental signaling pathway in bone formation as well as bone tumor pathophysiology. The aim of the present study was to evaluate the impact of the expression of RANK and its downstream signaling molecule Akt1 on tumor progression in patients with osteosarcoma. Expression of RANK and Akt1 was examined in 78 human osteosarcoma samples by immunohistochemistry using formalin-fixed samples. Following this, each graded immunohistochemistry result was correlated with clinicopathological parameters and patient survival. In total, 60 osteosarcomas (76.9%) expressed RANK and 58 cases (74.4%) showed expression of Akt1. In addition, expression of RANK was negatively correlated with disease-free survival by Kaplan-Meier analysis. A resistance was observed to chemotherapy in RANK-expressing cases, which was statistically significant (P<0.05). In addition, chemotherapy and staging of the tumor were found to independent factors that have an effect on patient survival (P<0.05). Thus, RANK was identified as a negative prognostic factor of osteosarcoma survival.

A Three-Dimensional Finite Element Analysis of Displaced Intra-Articular Calcaneal Fractures.

A better understanding of displaced intra-articular calcaneal fractures, their effect on joint mechanics, and the relationship between altered mechanics and osteoarthritis could aid in the development or refinement of treatment methods. Finite element modeling is accepted as the reference standard for evaluating joint contact stresses. The objective of the present study was to analyze the in vivo joint mechanical data from finite element modeling for normal and injured subtalar joints. A 3-dimensional model of the ankle-hindfoot was developed and validated. Both height loss and width increases in the calcaneus were simulated. Next, they were used to investigate the relationship between calcaneal height or width and the contact mechanics of the posterior facet of the subtalar joint. The contact area/joint area ratio increased in the subtalar joint with injury when the calcaneal width increased. Moreover, the peak contact pressure and the proportion of the area under high contact pressure (>6 MPa) increased. The contact area/joint area ratio decreased with reduced calcaneal height, but the peak contact pressure remained almost constant. The width increases of the calcaneus somewhat limited the subtalar joint motion, especially for eversion; however, the height loss mostly resulted in subtalar rotatory instability. The height loss diminished the subtalar joint's stability in eversion, internal rotation, and external rotation. The results of the present study support the advisability of surgery for these complex injuries. Reestablishing the calcaneal height and width could restore the normal kinematics and contact stress distribution in the subtalar joint, improve the tibiotalar position, and diminish long-term degeneration in the ankle.

Effect of Posterior Cruciate Ligament Rupture on Biomechanical and Histological Features of Lateral Femoral Condyle.

BACKGROUND The aim of this study was to investigate bone mineral density (BMD) and the biomechanical and histological effects of posterior cruciate ligament (PCL) rupture on the lateral femoral condyle. MATERIAL AND METHODS Strain on different parts of the lateral femoral condyle from specimens of normal adult knee joints, including 12 intact PCLs, 6 ruptures of the anterolateral bundle, 6 ruptures of the postmedial bundle, and 12 complete ruptures, was tested when loaded with different loads on the knee at various flexion angles. Lateral femoral condyles were also collected randomly from both the experimental side in which the PCLs were transected and the control side from 4 sets of 12 matched-mode pairs of rabbits at 4, 8, 16, and 24 weeks after surgery, and their BMD and morphological and histological changes were observed. RESULTS Partial and complete rupture of the PCL may cause an abnormal load on all parts of the lateral femoral condyle with any axial loading at all positions. Noticeable time-dependent degenerative histological changes of the lateral femoral condyle were observed in the rabbit model of PCL rupture. All of the PCL rupture groups had a higher expression of matrix metalloproteinase-7 (MMP-7) and collagen type II than the control group at all time points (P<0.05), but no significant difference in BMD (P>0.05). CONCLUSIONS Rupture of the PCL may trigger a coordinated response of lateral femoral condyle degeneration in a time-dependent manner, to which the high level of expression of MMP-7 and collagen type II could contribute.

Visfatin triggers the in vitro migration of osteosarcoma cells via activation of NF-κB/IL-6 signals.

Pulmonary metastasis is the major challenge for clinical treatment of osteosarcoma patients. Recent studies indicated that visfatin, a 52kDa adipocytokine, can trigger the cell motility of various cancers, while its role in the progression of osteosarcoma remains not clear. Our present study revealed that visfatin can significantly promote the in vitro migration and invasion of osteosarcoma MG-63 and HOS cells and up regulate the expression of matrix metalloproteinase-2 (MMP-2) and fibronectin (FN). Furthermore, visfatin treatment also increased the expression of IL-6 in both MG-63 and HOS cells via a time dependent manner, while anti-IL-6 antibody can significantly attenuate visfatin induced cell invasion and up regulation of MMP-2 and FN. It suggested that up regulation of IL-6 mediated visfatin induced in vitro motility of osteosarcoma cells. Visfatin treatment can increase the phosphorylation of both p65 and ERK1/2 in MG-63 and HOS cells, while only the inhibitor of NF-κB, BAY 11-7082, can abolish visfatin induced up regulation of IL-6. BAY 11-7082 also attenuated visfatin induced upregulation of MMP-2 and FN in MG-63 cells. Western blot analysis revealed that visfatin treatment can significantly increase the phosphorylation of IκBα and IKKß in MG-63 cells. ACHP, the inhibitor of IKK-ß, blocked visfatin induced expression of IL-6 mRNA in both MG-63 and HOS cells. Collectively, our data suggested that visfatin can increase the motility of osteosarcoma cells via up regulation of NF-κB/IL-6 signals. It indicated that visfatin might be a potential therapeutic target of osteosarcoma treatment.

miR-145 promotes osteosarcoma growth by reducing expression of the transcription factor friend leukemia virus integration 1.

Osteosarcoma (OS) is the most common malignant bone tumor in children and young adults. miR-145 is a microRNA highly expressed in vascularized tissues and has been widely studied in cancers. In this study, we explored the expression and function of miR-145 in OS. We found that miR-145 was consistently under-expressed in OS tissues and cell lines as compared to normal bone tissues and osteoblast cells. Ectopic expression of miR-145 in OS cells inhibited their proliferation and migration and induced apoptosis. miR-145 targets a putative microRNA regulatory element (MRE) in the 3'-UTR of friend leukemia virus integration 1 gene (FLI-1), and its abundance was inversely related to FLI-1 expression in OS tissues and cell lines. miR-145 decreased expression FLI-1 protein and mRNA, but mutation of the miR-145 MRE sequence in the FLI-1 3'-UTR abolished the activity of miR-145 in a reporter assay. Restored expression of FLI-1 diminished miR-145-mediated suppression of tumor progression. These results suggest that miR-145 acts as a tumor suppressor by directly reducing expression of FLI-1, and that the miR-145/FLI-1 pathway is important for tumor progression in OS.

Reconstruction of foot and ankle defects with a superthin innervated anterolateral thigh perforator flap.

BACKGROUND: An anterolateral thigh (ALT) perforator flap can be thinned to an extent to which it is vascularised only by the subdermal plexus. This study presents an innervated flap thinning technique and its application for dorsal foot and ankle resurfacing. METHODS: A superthin innervated ALT perforator flap was used to repair the dorsal foot and ankle of 12 patients. The perforators were classified according to their variations in the adipose layer, and the corresponding microdissection technique was then applied. The branch of the lateral femoral cutaneous nerve and its accompanying vessels were adopted to construct a sensory flap. RESULTS: The flap thickness before defatting, which was measured immediately after flap elevation, ranged from 25-45 mm. The average flap thickness after defatting was 4.55 mm (range = 3-6 mm). A total of 11 flaps completely survived, and one flap presented superficial necrosis within a small area (2 cm ×2 cm) in the distal part of the flap. No further flap revision or defatting procedures were required for these patients during an average follow-up period of 16.5 months (range = 10-24 months). In the transferred flap, protective sensibility existed in all cases, and the static two-point discrimination was 13-16 mm. CONCLUSIONS: The superthin innervated ALT perforator flap may be considered as an ideal strategy for foot and ankle reconstruction.

MiR-125b inhibits stromal cell proliferation in giant cell tumor of bone by targeting parathyroid hormone 1 receptor.

OBJECTIVES: miR-125b has been identified as a tumor suppressor in many tumors, but its role in giant cell tumor (GCT) of bone remains poorly understood. The current study aimed to investigate the potential role and mechanism of miR-125b in GCT. MATERIALS AND METHODS: Expression levels of miR-125b in GCT tissues were determined using RT-PCR. The cell proliferation was surveyed by direct cell counting, MTS and CCK-8, and the apoptotic cells were evaluated by Annexin V-FITC and propidium iodine staining assay. The target gene expression was determined using RT-PCR and western blot. Parathyroid hormone 1 receptor (PTH1R) 3'-UTR was cloned into luciferase reporter plasmid to confirm direct targeting. RESULTS: We found that miR-125b was significantly down-regulated in GCT tissues. Using both gain- and loss-of-function analyses, we further revealed that miR-125b suppressed GCT stromal cell proliferation and induced cell apoptosis. Furthermore, we revealed that PTH/PTHrP type 1 receptor is a direct and functional target of miR-125b. CONCLUSION: Our results suggest that miR-125b acts as a tumor suppressor through suppression of the PTH1R/RANKL signaling pathway. These findings contribute to our understanding of the functions of miR-125b in GCT.

Tenuigenin inhibits RANKL-induced osteoclastogenesis by down-regulating NF-κB activation and suppresses bone loss in vivo.

Tenuigenin, a major active component of polygala tenuifolia root, has been used to treat patients with insomnia, dementia, and neurosis. In this study, we aimed to investigate the effects of tenuigenin on osteoclastogenesis and clarify the possible mechanism. We showed that tenuigenin inhibited receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation and bone resorption without cytotoxicity, which was further demonstrated by reduced osteoclast specific gene expression such as TRAP, c-Src, ATP6v0d2, etc. Moreover, the inhibitory effect of tenuigenin was associated with impaired NF-κB activity owing to delayed degradation/regeneration of IkBa and inhibition of p65 nuclear translocation. Consistent with the in vitro results, micro-ct scanning and analysis data showed that tenuigenin suppressed RANKL-induced bone loss in an animal model. Taken together, our data demonstrate that tenuigenin inhibit osteoclast formation and bone resorption both in vitro and in vivo, and comprise a potential therapeutic alternative for osteoclast-related disorders such as osteoporosis and cancer-induced bone destruction.

MicroRNA-498 Inhibition Enhances the Differentiation of Human Adipose-Derived Mesenchymal Stem Cells into Podocyte-Like Cells.

Podocyte depletion is a key event in the progression of end-stage kidney disease (ESKD) resulting in nephrotic proteinuria and renal failure, but the treatment options are limited to dialysis and renal transplantation. So there is an urgent need for renal regenerative therapies. Generation of podocytes from human stem cells is regarded as a promising therapeutic strategy to repair or regenerate the damaged kidneys; however, the reliable induction system remains a challenge. In this study, we established a two-stage induction protocol for podocyte generation from human adipose-derived mesenchymal stem cells (hAD-MSCs). We initially established a condition that induces hAD-MSCs toward intermediate mesoderm cells with activin A and high concentration of retinoic acid (RA). Subsequently, by using the combination of activin A and low concentration of RA and BMP7, we generated podocyte-like cells expressing multiple podocyte-specific markers and able to integrate into a developing nephron of embryonic kidney explant culture and ameliorate proteinuria and kidney injure in adriamycin-treated mice. Furthermore, we identified that miRNA-498 inhibitor has potential to improve the differentiation of hAD-MSCs into podocyte-like cells and established a robust induction protocol. Thereby, our study advocated an efficient method for the induction of kidney podocyte-like (iPod) cells from hAD-MSCs and provided an ideal candidate for regenerative therapies of the kidney.

Downregulation of miR-22 acts as an unfavorable prognostic biomarker in osteosarcoma.

miRNA-22 (miR-22) has been showed to involve in a variety of cancers; however, the association between miR-22 expression level and the prognosis of osteosarcoma is also poorly unknown. Fifty-two patients with surgically resected paired osteosarcoma and non-neoplastic disease between 2008 and 2014 were involved in this study. Real-time PCR was performed to examine the expression level of miR-22 in osteosarcoma tissues and noncancerous bone tissues. Then the association between miR-22 expression and clinical-pathological parameters were further evaluated. Kaplan-Meier analysis and Cox proportional hazards regression models were explored to reveal the correlations of miR-22 expression with survival of patients. The results indicated that miR-22 was downregulated in osteosarcoma tissues in comparison with noncancerous bone tissues. In addition, there is statistically significance between miR-22 expression level and recurrence, metastasis, and chemotherapy response. The patients with lower miR-22 expression level had both poorer overall survival and disease-free survival. The multivariant analysis revealed that the miR-22 expression level and metastasis status are independent prognosis factors for osteosarcoma. In conclusion, miR-22 was downregulated in osteosarcoma and its expression level was correlated with a variety of important clinical-pathological parameters. Moreover, miR-22 may serve as a promising biomarker for predicting the prognosis of osteosarcoma.

Benefit of degenerative posterior longitudinal ligament removal during anterior decompression in cervical spondylotic myelopathy.

It remains controversial whether degenerative posterior longitudinal ligaments should be removed during anterior decompression procedures for cervical spondylotic myelopathy. Few data are available from studies that have compared removing and retaining the degenerative posterior longitudinal ligament. The goal of this retrospective study was to evaluate the benefit of degenerative posterior longitudinal ligament removal during such operations. Clinical data on 130 patients with confirmed degenerative posterior longitudinal ligament who underwent anterior cervical decompression surgery were retrospectively reviewed. All procedures were performed by the same senior orthopedic surgeon at the authors' spinal surgery center. The degenerative posterior longitudinal ligament was removed in 62 patients (group A) and retained in 68 patients (group B). The 130 patients were followed for 36 months. The Japanese Orthopedic Association score improved from 9.0±2.7 to 14.7±1.5 in group A and from 9.4±2.6 to 14.1±1.7 in group B (P=.028). The recovery rate for spinal cord neurologic function was 66.7% in group A and 61.3% in group B (P=.031). Operating time was longer (P=.002) and the sagittal median diameter of the vertebral canal was enhanced in group A (P<.001). Narrowing of previously enlarged canals occurred at a significantly higher rate in group B (P=.044). No significant difference was found in the rates of common complications between groups. Removal of the degenerative posterior longitudinal ligament in anterior decompression procedures for cervical spondylotic myelopathy appeared to be beneficial and provided more complete decompression and better postoperative outcomes than surgery without removal of the ligament. Although this procedure was generally safe, it required longer operating times, was more technically challenging, and required more experienced surgeons than surgery without removal of the ligament.

RANK pathway in giant cell tumor of bone: pathogenesis and therapeutic aspects.

Giant cell tumor is a relatively uncommon but painful tumor of bone, which can metastasize to the lungs. The RANK pathway is often reported to be involved in the pathogenesis of giant cell tumor of bone (GCTB). This pathway is a key signaling pathway of bone remodeling that plays a critical role in differentiation of precursors into multinucleated osteoclasts, and activation of osteoclasts leading to bone resorption. Dysregulation of RANK ligand (RANKL)-RANK-osteoprotegerin (OPG) signaling cascade induces the imbalance between bone formation and bone resorption, which leads to the changes in bone mass, increases osteoclast-mediated bone destruction, bone metastasis, and the progression of existing skeletal tumors. Recent evidences have shown that targeting the components of RANKL-RANK-OPG signaling pathway is a promising approach in the treatment of GCTB. This review study has focused on the association of RANKL-RANK-OPG pathway in the pathogenesis and progression of GCTB as well as discussed the possible therapeutic strategies by targeting this pathway.

Morphological changes and expression of MMPs and TIMPs in rabbit degenerated lateral meniscus after PCL-transection.

INTRODUCTION: Whether the expression level of MMP-1, MMP-13 and TIMP-1 has association with the degeneration of lateral meniscus after posterior cruciate ligament (PCL) fracture is poorly understood. The aim of this study was to investigate the influence of PCL fracture on lateral meniscus, including morphological changes, histological changes and roles of matrix metalloproteinase-1 (MMP-1), matrix metalloproteinase-13 (MMP-13) and tissue inhibitor of metalloproteinase-1 (TIMP-1) expression level in the secondary injury. MATERIALS AND METHODS: Sixty male rabbits were used as PCL transection models and randomized into the PCL-transection side, which underwent PCL transection surgery, and the control side, which underwent PCL exposure without transection. On 4, 8, 12, 16 and 24 weeks after PCL-transection, 12 rabbits were randomly killed for H&E staining to determine the histological changes of lateral meniscus. Immunohistochemical staining was undertaken to evaluate the expression level of MMP-1, MMP-13 and TIMP-1 in lateral meniscus. The results were statistically analyzed using SPSS 15.0. RESULTS: The lateral meniscus of PCL-transection side presented abnormal morphology. Histological evaluation score of meniscal degeneration in PCL-transection group was higher than that in the control group with statistical difference (P < 0.05). The expression levels of MMP-1, MMP-13 and TIMP-1 were significantly elevated in meniscus of the PCL-transection group with statistical difference (P < 0.05). MMP-1 expression displayed an increasing trend firstly then kept stable after PCL transection; MMP-13 and TIMP-1 expression displayed high level firstly then decreased in advanced stage after PCL transection. CONCLUSIONS: PCL transaction may induce a coordinated response of degeneration of lateral meniscus in a time-dependent manner. The high expression level of MMP-1, MMP-13 and TIMP-1 would contribute to the degeneration of lateral meniscus after PCL transection.

Inhibition of osteosarcoma cell progression by MacroH2A via the downregulation of cyclin D and cyclin­dependent kinase genes.

MacroH2A is a histone modification factor the activity of which has been acutely studied in cancer progression, and a number of studies have shown that the progression of certain types of cancer is under regulation by MacroH2A. However, information regarding the underlying molecular mechanisms of MacroH2A inhibition on the cell cycle remains elusive, and elucidating this process may aid in the production of novel treatment strategies. The aim of the current study was to investigate the inhibitory effects of MacroH2A on osteosarcoma cell progression, and the possible molecular mechanisms of this process. MacroH2A overexpression and interference vectors were designed and transfected into U2­OS osteosarcoma cells. The cells underwent reverse transcription­quantitative polymerase chain reaction (RT­qPCR), western blot analysis and immunofluorescence assays. The apoptosis rate and cell cycle stage were assayed using flow cytometry. The results revealed that the overexpression of MacroH2A inhibited the progression of U2­OS osteosarcoma cells, and the cells were arrested at the G2/M stage of the cell cycle. The molecular mechanism by which MacroH2A suppresses the cell progression involves the inhibition of the expression of cyclin D and cyclin­dependent kinase (CDK) genes, including cyclin D1, cyclin D2, CDK4, CDK6 and CDK8. Taken together, the present results revealed that MacroH2A is an important modifier of chromatin that downregulates the progression of osteosarcoma cells and triggers disturbance of the cell cycle via the downregulation of cyclin D and CDK genes.

Role of aldolase A in osteosarcoma progression and metastasis: in vitro and in vivo evidence.

Aldolase A (ALDOA) has been reported to be negative survival marker of osteosarcoma (OS) and may be implicated in OS development and progression. In the present study, we assessed for the first time the functional role of ALDOA in OS cell invasion and survival in vitro and in vivo, using human OS cell lines and an orthotopic xenograft nude mouse model. Overexpression and knockdown of ALDOA were respectively performed in MG-63 and U-2 OS cells, which showed relatively low and high constitutive ALDOA expression levels, respectively. Overexpression of ALDOA in MG-63 cells significantly increased in vitro cell invasion, matrix metalloproteinase (MMP)-2 expression, and cell survival against cisplatin-induced apoptosis. On the other hand, knockdown of ALDOA in U-2 cells markedly decreased in vitro cell invasion, MMP-2 expression, and cell survival against cisplatin-induced apoptosis. In an orthotopic xenograft nude mouse model, intra-tibial injection of MG-63 cells overexpressing ALDOA led to significantly increased primary tumor volume and pulmonary metastasis as well as decreased cell apoptosis in the primary tumors, compared with the controls. In contrast, intra-tibial injection of U-2 cells with knockdown of ALDOA led to markedly decreased primary tumor volume and pulmonary metastasis as well as increased cell apoptosis in the primary tumors, compared with the controls. In conclusion, our in vitro data indicate that ALDOA promotes OS cell invasion and survival, and our in vivo data demonstrate an important role of ALDOA in promoting OS tumor growth and metastasis. The present study provides the first in vitro and in vivo evidence supporting a critical functional role of ALDOA in OS progression and metastasis, suggesting that ALDOA could serve as a novel therapeutic target in OS. Additionally, our results suggest that ALDOA is involved in the development of OS chemoresistance.