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Background: The accuracy of pedicle screw fixation is crucial for patient safety. Traditional navigation methods based on computed tomography (CT) imaging have several limitations. Therefore, this study aimed to investigate the ultrasonic propagation characteristics of bone tissue and their relationship with CT imaging results, as well as the potential application of ultrasound navigation in pedicle screw fixation. Methods: The study used three bovine spine specimens (BSSs) and five human vertebral allograft bones (HABs) to progressively decrease the thickness of the cancellous bone layer, simulating the process of pedicle screw perforation. Five unfocused miniature ultrasound probes with frequencies of 2.2, 2.5, 3, 12, and 30 MHz were employed for investigating the ultrasonic propagation characteristics of cancellous and cortical bone through ultrasound transmission and backscatter experiments. The CT features of the bone tissue was obtained with the Skyscan 1174 micro-CT scanner (Bruker, Billerica, MA, USA). Results: The experimental results demonstrated that low-frequency (2-3 MHz) ultrasound effectively penetrated the cancellous bone layer up to a depth of approximately 5 mm, with an attenuation coefficient below 10 dB/cm. Conversely, high-frequency (12 MHz) ultrasound exhibited significant signal attenuation in cancellous bone, reaching up to 55.8 dB/cm. The amplitude of the backscattered signal at the cancellous bone interface exhibited a negative correlation with the bone sample thickness (average r=-0.84), meaning that as the thickness of the cancellous bone layer on the cortical bone decreases, the backscattered signal amplitude gradually increases (P<0.05). Upon reaching the cortical bone interface, there was a rapid surge in echo signal amplitude, up to 8 times higher. Meanwhile, the statistical results indicated a significant correlation between the amplitude of the echo signal and the micro-CT scanning results of bone trabecular structure. Conclusions: Theoretically, using multiple ultrasonic probes (≥3) and regions of interest (ROIs) (≥5) has the potential to provide surgeons with early warning signals for pedicle perforation based on three or more successive increases in echo signal amplitude or a sudden substantial increase. The statistical results indicate a significant correlation between the amplitude of the echo signal and the micro-CT scanning results of bone trabeculae, suggesting the potential use of ultrasound as opposed to CT for real-time intraoperative bone navigation.
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BACKGROUND Arthritis of the hip caused by arteriovenous malformations (AVMs) has been rarely reported. Therefore, total hip replacement (THR) in patients with AVM-induced arthritis of the hip is challenging. CASE SUMMARY We report a 44-year-old woman with aggravated right hip pain during the past decade. The patient presented with severe pain and a functional disorder of the right hip. X-ray examination revealed severely narrowed right hip joint space and abnormal trabecular bone loss in the femoral neck and trochanter area. Doppler ultrasound, magnetic resonance imaging and computed tomography angiography revealed AVMs surrounding the right hip, along with erosion. To ensure the safety of THR, we performed vascular embolization and temporary balloon occlusion of the iliac artery three times during the operation. However, serious hemorrhage occurred, which was rescued by the multimodality blood conservation strategy. THR was successfully performed, and the patient was discharged 8 d later for rehabilitation. Postoperative pathological examination showed osteonecrosis of the femoral head with malformed thick-walled vessels and focal granulomatous inflammation of the surrounding soft tissues. The Harris Hip Scale score increased from 31 to 82 at 3 mo of follow-up. The patient was followed up for 1 year, and all her clinical symptoms were significantly alleviated. CONCLUSION Arthritis of the hip caused by AVMs is rare in clinical practice. The activity and function of the involved hip joint can be effectively treated with THR after comprehensive imaging and multidisciplinary consultation.
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Study design: Retrospective study. Objectives: The traditional PLIF is routinely utilized in severe lumbar spinal stenosis to relief the nerve compression. Nevertheless, the removal of posterior tension-band structure and the denervation and atrophy of the paraspinal muscle affect the clinical efficacy. Therefore, unilateral modified PLIF combined with contralateral fenestration was performed to overcome above-mentioned drawbacks. Methods: 32 modified PLIF and 33 traditional PLIF cases were retrospectively included. Operation time, length of stay (LOS) and blood loss were recorded. VAS of low back pain and leg pain, ODI and Sf-36 score including physical function and body pain were assessed. Fusion rate, lumbar lordosis (LL), intervertebral angle (IVA) and intervertebral height index (IHI) were evaluated radiologically. Results: Modified group possessed less blood loss, shorter operation time and less LOS. Compared with traditional group, the VAS of back pain was lower at 6 months postoperatively (P < .05) and the ODI score was lower at 3 months postoperatively (P < .05) in modified group. Modified group exhibited better physical function 3 months postoperatively and lower body pain 6 months postoperatively in Sf-36 score (P < .05). No statistic difference in LL, IVA, IHI and fusion rate were observed between both groups. Conclusions: Our modified PLIF combining with contralateral fenestration procedure exhibited particular advantages in comparison to traditional PLIF. The preservation of posterior tension-band structure facilitates to less low back pain, low complication rate and early functional recovery.
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Fusión Vertebral , Humanos , Estudios Retrospectivos , Constricción Patológica , Vértebras Lumbares/diagnóstico por imagen , Vértebras Lumbares/cirugía , Resultado del Tratamiento , DolorRESUMEN
BACKGROUND: The accessory bones are common bone variations around the feet and ankles, which usually originate from nonunion of the secondary ossification center adjacent to the main bone mass, and most of them remain asymptomatic. Os subcalcis is an accessory bone at the plantar aspect of the calcaneus, which is located just posterior to the insertion of the plantar fascia. Focal bone formation at the calcaneal plantar pole with heel pain has rarely been reported. CASE SUMMARY: A 55-year-old man presented to our clinic with left plantar heel pain and a progressive swelling for 8 years. X-ray, computer tomography and magnetic resonance imaging showed a large os subcalcison the plantar side of the calcaneus, located at the insertion of the plantar fascia. He underwent surgical excision of the lesion. Microscopically the bony trabeculae were intermingled with fat and covered with cartilage. CONCLUSION: This is a rare case with accessory os subcalcis leading to heel pain. It highlights the awareness of os subcalcis and helps avoid future misdiagnosis of heel pain.
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Signal transducer and activator of transcription 3 (STAT3) plays essential roles in cancer progression and has been considered as a promising target for cancer therapy. Here, we used a dual luciferase assay to identify that pectolinarigenin inhibited STAT3 transcriptional activity. Further, results showed pectolinarigenin inhibited constitutive and IL6 induced STAT3 signaling, diminished the accumulation of STAT3 in the nucleus, dimerization and blocked STAT3 DNA binding activity. Mechanism investigations indicated that pectolinarigenin disturbed the STAT3/DNMT1/HDAC1 complex formation in the promoter region of SHP-1, which reversely mediates STAT3 signaling, leading to the upregulation of SHP-1 expression in osteosarcoma. We also found pectolinarigenin significantly suppressed osteosarcoma growth, induced apoptosis. In addition, pectolinarigenin blocked tumor cells migration, invasion and reserved EMT phenotype. In spontaneous tibial injection and patient-derived xenograft models of osteosarcoma, we identified administration (i.p.) of pectolinarigenin (20 mg/kg/2 days and 50 mg/kg/2 days) blocked STAT3 activation and disturbed tumor growth and metastasis with superior pharmacodynamic properties. Taken together, our findings demonstrate that pectolinarigenin may be a candidate for osteosarcoma intervention linked to its STAT3 signaling inhibitory activity.
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Neoplasias Óseas , Osteosarcoma , Apoptosis , Neoplasias Óseas/tratamiento farmacológico , Neoplasias Óseas/metabolismo , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Cromonas , Humanos , Janus Quinasa 2/metabolismo , Osteosarcoma/tratamiento farmacológico , Osteosarcoma/patología , Proteína Tirosina Fosfatasa no Receptora Tipo 6 , Factor de Transcripción STAT3/metabolismoRESUMEN
Increased adipocyte and decreased osteoblast differentiation, combined with the ectopic proliferation of bone marrow mesenchymal stem cells (BM-MSCs), represent the primary causes of osteoporosis. The dysregulation of numerous intracellular bioactive factors is responsible for the aberrant differentiation and growth of BM-MSCs. In this study, we focused on a new stimulative factor, integrator complex subunit 7 (INTS7), and its cooperative protein ATP-binding cassette subfamily D member 3 (ABCD3)/high-density lipoprotein-binding protein (HDLBP) in mouse BM-MSCs. We aimed to uncover the effects of the INTS7-ABCD3/HDLBP interaction on BM-MSC biological behaviors and the potential mechanism underlying these effects. Functional in vitro experiments showed that the suppression of the INTS7-ABCD3 interaction rather than HDLBP could impair BM-MSC proliferation and induce cell apoptosis. Moreover, Alizarin Red S and Oil Red O staining, respectively, revealed that INTS7 and ABCD3 knockdown but not HDLBP knockdown could decrease osteoblastic differentiation and accelerate the adipogenic differentiation of BM-MSCs. Mechanistically, reactive oxygen species (ROS) and histone γ-H2AX quantities significantly increased, whereas the levels of antioxidants declined due to INTS7 and ABCD3 inhibition in BM-MSCs. These findings indicated that the suppression of oxidative stress could be involved in the INTS7/ABCD3 co-regulatory mechanisms for BM-MSC proliferation and differentiation, identifying new potential candidates for osteoporosis therapy.
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BACKGROUND: The most commonly ingested foreign body in Asians is fish bone. The vast majority of patients have obvious symptoms and can be timely diagnosed and treated. Cases of pyogenic cervical spondylitis and diskitis with retropharyngeal and epidural abscess resulting in incomplete quadriplegia due to foreign body ingestion have been rarely reported. The absence of pharyngeal or esophageal discomfort and negative computed tomography (CT) findings of fish bone have not been reported. We report the case of an elderly female patient with delayed cervical infection and incomplete quadriplegia who had a history of fish bone ingestion. CASE SUMMARY: A 73-year-old woman presented with right neck pain and weakness of four limbs for a week, and had a history of fish bone ingestion and negative findings on laryngoscopic examination one month previously. She did not complain of any pharyngeal or esophageal discomfort. Cervical magnetic resonance imaging showed C4/C5 spondylitis and diskitis along with retropharyngeal and ventral epidural abscesses. No sign of fish bone was detected on lateral cervical radiography and CT scans. The muscle strength of the patient's right lower limb receded to grade 1 and other limbs to grade 2 suddenly on the 10th day of hospitalization. Emergency surgery was performed to drain the abscess and decompress the spinal cord by removing the anterior inflammatory necrotic tissue. Simultaneously, flexible esophagogastroduodenoscopy was carried out and a hole in the posterior pharyngeal wall was found. The motor weakness of the right lower limb improved to grade 3 and the other limbs to grade 4 within 2 d postoperatively. CONCLUSION: This rare case highlights the awareness of the posterior pharyngeal or esophageal wall perforation in patients with cervical pyogenic spondylitis along with a history of fish bone ingestion, even though local discomfort symptoms are absent and the radiological examinations are negative.
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Transcription factor brachyury, with a DNA-binding T-domain, regulates posterior mesoderm formation and notochord development through binding with highly conserved palindromic consensus sequence in a variety of organisms. The absence of brachyury expression in majority of adult normal tissues and exclusive tumor-specific expression provides the potential to be developed into a novel and promising diagnostic and therapeutic target in cancer. As a sensitive and specific marker in the diagnosis of chordoma, brachyury protein has been verified to involve in the process of carcinogenesis and progression of chordoma and several epithelial carcinomas in various studies, but the mechanism by which brachyury promotes tumor cells migrate, invade and metastasis still remains less clear. To this end, we attempt to summarize the literature on the upstream regulatory pathway of brachyury transcription and downstream controlling network by brachyury activation, all of which involve in both the embryonic development and tumor progression. We present the respective correlation of brachyury expression with tumor progression, distant metastasis, survival rate and prognosis in several types of tumor samples (including chordoma, lung cancer, breast carcinoma, and prostate cancer), and various brachyury gain-of-function and loss-of-function experiments are summarized to explore its specific role in respective tumor cell line in vitro. In addition, we also discuss another two programs relating to brachyury function: epithelial-to-mesenchymal transition (EMT) and cell cycle control, both of which implicate in the regulation of brachyury on biological behavior of tumor cells. This review will provide an overview of the function of master transcriptional factor brachyury, compare the similarities and differences of its role between embryonic development and carcinogenesis, and list the evidence on which brachyury-target therapies have the potential to help control advanced cancer populations.
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[This corrects the article DOI: 10.7150/thno.18299.].
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The bone marrow has been long known to host a unique environment amenable to colonization by metastasizing tumor cells. Yet, the underlying molecular interactions which give rise to the high incidence of bone metastasis (BM) in breast cancer patients have long remained uncharacterized. In our study, in vitro and in vivo assays demonstrated that Brachyury (Bry) could promote breast cancer BM. Bry drives epithelial-mesenchymal transition (EMT) and promotes breast cancer aggressiveness. As an EMT driver, SOX5 involves in breast cancer metastasis and the specific function in BM. Chromatin immunoprecipitation (ChIP) assays revealed SOX5 is a direct downstream target gene of Bry. ChIP analysis and reporter assays identified two Bry-binding motifs; one consistent with the classic conserved binding sequence and the other a new motif sequence. This study demonstrates for the first time that Bry promotes breast cancer cells BM through activating SOX5. In clinical practice, targeting the Bry-Sox5-EMT pathway is evolving into a promising avenue for the prevention of bone metastatic relapse, therapeutic resistance and other aspects of breast cancer progression. Brachyury directly regulates the expression of SOX5 by binding to two motifs in its promoter region. The Bry-SOX5-EMT pathway may represent a potential target to develop treatments to prevent and treat bone metastasis from breast cancer.
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Biomarcadores de Tumor/metabolismo , Neoplasias Óseas/secundario , Neoplasias de la Mama/patología , Proteínas Fetales/metabolismo , Regulación Neoplásica de la Expresión Génica , Factores de Transcripción SOXD/genética , Proteínas de Dominio T Box/metabolismo , Activación Transcripcional , Animales , Apoptosis , Biomarcadores de Tumor/genética , Neoplasias Óseas/genética , Neoplasias Óseas/metabolismo , Neoplasias de la Mama/genética , Neoplasias de la Mama/metabolismo , Adhesión Celular , Movimiento Celular , Proliferación Celular , Progresión de la Enfermedad , Transición Epitelial-Mesenquimal , Femenino , Proteínas Fetales/genética , Humanos , Ratones , Ratones Desnudos , Recurrencia Local de Neoplasia/genética , Recurrencia Local de Neoplasia/metabolismo , Recurrencia Local de Neoplasia/patología , Pronóstico , Factores de Transcripción SOXD/metabolismo , Tasa de Supervivencia , Proteínas de Dominio T Box/genética , Células Tumorales Cultivadas , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
The aim of this study was to evaluate the safety and efficacy of percutaneous vertebroplasty (PVP) in Kambin triangle approach for the treatment of osteoporotic vertebral compression fractures (OVCFs).Between November 2017 and September 2018, 109 patients (144 vertebral bodies) with OVCFs, with a mean age of 76.7â±â9.9 years (55-96 years), underwent PVP in Kambin triangle approach. The time of operation, the volume of bone cement, the incidence of complication, the Visual Analog Scale (VAS) and Oswestry Disability Index (ODI) score, the position of puncture needles, and the spread of polymethylmethacrylate (PMMA) in vertebral body (VB) were recorded.All patients had been completed the operation successfully and were followed up 9.1â±â2.9 months. The average operation time of each VB was 24.0â±â3.5âminutes. The average volume of cement was 4.8â±â0.6âml. The mean VAS scores were 8.4â±â0.7 preoperatively, 1.6â±â0.6 at the first day postoperatively, and 1.2â±â0.6 at the last follow-up. The mean ODI scores were 70.97â±â7.73 preoperatively, 27.99â±â4.12 at the first day postoperatively, and 19.65â±â3.49 at the last follow-up. The position of puncture needles in the VB was: 119 vertebral puncture needles reached the midline, 15 were close to the midline, and 10 exceeded the midline. The spread of PMMA in the VB was: type 1 in 81 levels (56.3%), type 2 in 37 (25.7%), type 3 in 18 (12.5%), type 5 in 8 (5.5%), and no case in type 4. One case developed pneumothorax after operation. No other complications (hematoma, cement embolism, spinal cord, and nerve injury) occurred.Kambin triangle approach in PVP, which can deliver the puncture needle to the midline of VB easily and with excellent cement distribution, is a safe and effective method.
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Fracturas por Compresión/cirugía , Fracturas Osteoporóticas/cirugía , Fracturas de la Columna Vertebral/cirugía , Vertebroplastia/métodos , Anciano , Anciano de 80 o más Años , Cementos para Huesos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Tempo Operativo , Polimetil Metacrilato , Complicaciones Posoperatorias , Estudios Retrospectivos , Vertebroplastia/efectos adversosRESUMEN
Fullerene C60 nanocrystals (nano-C60) possess various attractive bioactivities, including autophagy induction and calcium/calmodulin-dependent protein kinase IIα (CaMKIIα) activation. CaMKIIα is a multifunctional protein kinase involved in many cellular processes including tumor progression; however, the biological effects of CaMKIIα activity modulated by nano-C60 in tumors have not been reported, and the relationship between CaMKIIα activity and autophagic degradation remains unclear. Herein, nano-C60 is demonstrated to elicit reactive oxygen species (ROS)-dependent cytotoxicity and persistent activation of CaMKIIα in osteosarcoma (OS) cells. CaMKIIα activation, in turn, produces a protective effect against cytotoxicity from nano-C60 itself. Inhibition of CaMKIIα activity by either the chemical inhibitor KN-93 or CaMKIIα knockdown dramatically promotes the anti-OS effect of nano-C60. Moreover, inhibition of CaMKIIα activity causes lysosomal alkalinization and enlargement, and impairs the degradation function of lysosomes, leading to autophagosome accumulation. Importantly, excessive autophagosome accumulation and autophagic degradation blocking are shown to play an important role in KN-93-enhanced-OS cell death. The synergistic anti-OS efficacy of KN-93 and nano-C60 is further revealed in an OS-xenografted murine model. The results demonstrate that CaMKIIα inhibition, along with the suppression of autophagic degradation, presents a promising strategy for improving the antitumor efficacy of nano-C60.
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Early aggressive metastasis of osteosarcoma (OS) leads to rapid progression and poor prognosis. Increasing evidence has demonstrated that long non-coding RNAs (lncRNAs) could serve as crucial regulators to modulate tumour metastasis. In this study, we reported the critical role of lncRNA TUG1 in determining OS metastasis. TUG1 was significantly upregulated in OS tissues and associated with tumour size, distant metastasis, TNM stage, and overall and recurrence-free survival, which further indicated poor prognosis. Furthermore, CAFs-derived TGF-ß could upregulate TUG1 expression, and the crosstalk between CAFs and OS cells induced TUG1 to promote OS cell metastasis. Dysregulated TUG1 expression could act as an miRNA "sponge" to competitively protect the HIF-1α mRNA 3'UTR from miR-143-5p. Our study emphasised the effects of TUG1 in OS and demonstrated a novel axis by which TUG1 regulated OS cell metastasis, angiogenesis, and proliferation in vivo and in vitro. Collectively, TUG1 might be a prognostic indicator for OS and could be a therapeutic target for OS.
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Neoplasias Óseas/metabolismo , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , MicroARNs/metabolismo , Osteosarcoma/metabolismo , ARN Largo no Codificante/metabolismo , Adolescente , Animales , Neoplasias Óseas/mortalidad , Neoplasias Óseas/patología , Línea Celular Tumoral , Femenino , Xenoinjertos , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Masculino , Ratones , Ratones Desnudos , Metástasis de la Neoplasia/genética , Osteosarcoma/mortalidad , Osteosarcoma/patología , Pronóstico , Interferencia de ARN , ARN Largo no Codificante/genética , Tasa de Supervivencia , Carga Tumoral/genética , Adulto JovenRESUMEN
Since publication of this article, the authors have noticed errors in Fig. 1e (the merge image of control group) and Fig. 5e (Pec. 50 mg/kg group). As a result of the misfiling of the data, incorrect images were inadvertently inserted in Figs. 1e and 5e during figure preparation. The correct figures are given below.
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PURPOSE: Osteosarcoma is the most common primary bone cancer and is notorious for pulmonary metastasis, representing a major threat to pediatric patients. An effective drug targeting osteosarcoma and its lung metastasis is urgently needed. DESIGN: In this study, a sarcoma-targeting peptide-decorated disulfide-crosslinked polypeptide nanogel (STP-NG) was exploited for enhanced intracellular delivery of shikonin (SHK), an extract of a medicinal herb, to inhibit osteosarcoma progression with minimal systemic toxicity. RESULTS: The targeted, loaded nanogel, STP-NG/SHK, killed osteosarcoma cells by inducing RIP1- and RIP3-dependent necroptosis in vitro. Necroptosis is a novel cell death form that could be well adapted as an efficient antitumor strategy, the main obstacle of which is its high toxicity. After intravenous injection, STP-NG/SHK efficiently suppressed tumor growth and reduced pulmonary metastasis, offering greater tumor necrosis and higher RIP1 and RIP3 upregulation compared to free SHK or untargeted NG/SHK in vivo. Additionally, the treatment with NG/SHK or STP-NG/SHK showed minimal toxicity to normal organs, suggesting low systemic toxicity compared to free SHK. CONCLUSION: The STP-guided intracellular drug delivery system using the necroptosis mechanism showed profound anti-osteosarcoma activity, especially eliminated lung metastasis in vivo. This drug formulation may have great potential for treatment of osteosarcoma.
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Apoptosis , Neoplasias Pulmonares/secundario , Naftoquinonas/administración & dosificación , Osteosarcoma/patología , Péptidos/química , Polietilenglicoles/química , Polietileneimina/química , Sarcoma/patología , Regulación hacia Arriba , Animales , Línea Celular Tumoral , Sistemas de Liberación de Medicamentos , Femenino , Humanos , Espacio Intracelular/metabolismo , Neoplasias Pulmonares/tratamiento farmacológico , Ratones Desnudos , Nanogeles , Naftoquinonas/efectos adversos , Naftoquinonas/uso terapéutico , Necrosis , Osteosarcoma/tratamiento farmacológico , Polietilenglicoles/efectos adversos , Polietileneimina/efectos adversos , Distribución TisularRESUMEN
Osteosarcoma is the most common primary malignant bone tumor. Cancer cells employ a host of mechanisms to develop resistance to adriamycin (ADM) or other chemotherapeutic drugs. Shikonin (SK), an active constituent extracted from a Chinese medicinal herb, has been shown to cooperate with ADM in the treatment of osteosarcoma and certain other types of cancer by contributing to the response rate of chemotherapy and the side effects. The aim of the present study was to investigate the role and underlying mechanism of SK in chemotherapy for osteosarcoma. In the present study, a CCK-8 assay was performed to assess cell survival rate in vitro. Western blot analysis was performed to determine the expression levels of Bcell lymphoma 2associated X protein (Bax), caspase3, caspase8, and poly (ADPribose) polymerase (PARP). Flow cytometry was used to analyze cell cycle and cell death. The survival rate of cells decreased significantly in a dose and timedependent manner when treated with a combination of SK and ADM. Western blot analysis revealed increased expression levels of Bax, caspase3, caspase8 and PARP in U2OS and MG63 cells 48 h following treatment with SK and ADM. Flow cytometric analysis showed that the combined treatment of SK and ADM significantly induced apoptosis in the osteosarcoma cells. Taken together SK cooperated with ADM to promote apoptosis, possibly by inducing caspase3 and caspase8dependent apoptosis. SK may be a potential enhancer in the treatment of drugresistant primary osteosarcoma.
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Antibióticos Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Neoplasias Óseas/metabolismo , Caspasa 3/metabolismo , Caspasa 8/metabolismo , Doxorrubicina/farmacología , Naftoquinonas/farmacología , Osteosarcoma/metabolismo , Neoplasias Óseas/genética , Caspasa 3/genética , Caspasa 8/genética , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Sinergismo Farmacológico , Humanos , Osteosarcoma/genéticaRESUMEN
Osteosarcoma (OS) is the most frequent primary malignant bone tumour. Alternol, a novel compound purified from microbial fermentation products exerts anti-tumour effects across several cancer types. The effect of alternol on human OS remains to be elucidated. We first evaluated the anti-tumour effect of alternol in several human OS cell lines in vitro and investigated its underlying mechanism. Alternol inhibited OS cell proliferation, migration and induced caspase-dependent apoptosis, G2/M cell cycle arrest in a dose and time-dependent manner. Moreover, alternol treatment inhibited signal transducer and activator of transcription-3 (STAT3) phosphorylation in 143B and MG63 human OS cells, as evaluated using a STAT3-dependent dual luciferase reporter system. Exposure to alternol resulted in excessive reactive oxygen species (ROS) generation and Jun amino-terminal kinases (JNK), extracellular signal-regulated kinases (ERK1/2) and p38 activation. Furthermore, alternol-induced cell death was significantly restored in the presence of the ROS scavenger, N-acetyl-l-cysteine (NAC) or a caspase inhibitor Z-VAD-FMK. NAC also prevented G2/M phase arrest and phosphorylation of mitogen-activated protein kinases (MAPK), but did not reverse STAT3 inactivation. Finally, alternol suppressed tumour growth in vivo in the nude mouse OS tibia orthotopic model. Immunohistochemistry revealed that alternol treatment resulted in down-regulation of phosph-STAT3 Tyr705 and up-regulation of cleaved caspase-3 and phosph-SAPK (Stress-activated protein kinases)/JNK expression. Taken together, our results reveal that alternol suppresses cell proliferation, migration and induces apoptosis, cell cycle arrest by modulating of ROS-dependent MAPK and STAT3 signalling pathways in human OS cells. Therefore, alternol is a promising candidate for developing anti-tumour drugs target OS.
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Antineoplásicos/uso terapéutico , Productos Biológicos/uso terapéutico , Compuestos Heterocíclicos de 4 o más Anillos/uso terapéutico , Sistema de Señalización de MAP Quinasas , Osteosarcoma/tratamiento farmacológico , Especies Reactivas de Oxígeno/metabolismo , Factor de Transcripción STAT3/metabolismo , Animales , Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Productos Biológicos/farmacología , Caspasas/metabolismo , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Puntos de Control de la Fase G2 del Ciclo Celular/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Compuestos Heterocíclicos de 4 o más Anillos/química , Compuestos Heterocíclicos de 4 o más Anillos/farmacología , Humanos , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Ratones Endogámicos BALB C , Ratones Desnudos , Invasividad Neoplásica , Osteosarcoma/enzimología , Osteosarcoma/patología , Factores de TiempoRESUMEN
Signal transducer and activator of transcription 3 (STAT3) has important roles in cancer aggressiveness and has been confirmed as an attractive target for cancer therapy. In this study, we used a dual-luciferase assay to identify that pectolinarigenin inhibited STAT3 activity. Further studies showed pectolinarigenin inhibited constitutive and interleukin-6-induced STAT3 signaling, diminished the accumulation of STAT3 in the nucleus and blocked STAT3 DNA-binding activity in osteosarcoma cells. Mechanism investigations indicated that pectolinarigenin disturbed the STAT3/DNA methyltransferase 1/HDAC1 histone deacetylase 1 complex formation in the promoter region of SHP-1, which reversely mediates STAT3 signaling, leading to the upregulation of SHP-1 expression in osteosarcoma. We also found pectolinarigenin significantly suppressed osteosarcoma cell proliferation, induced apoptosis and reduced the level of STAT3 downstream proteins cyclin D1, Survivin, B-cell lymphoma 2 (Bcl-2), B-cell lymphoma extra-large (Bcl-xl) and myeloid cell leukemia 1 (Mcl-1). In addition, pectolinarigenin inhibited migration, invasion and reserved epithelial-mesenchymal transition (EMT) phenotype in osteosarcoma cells. In spontaneous and patient-derived xenograft models of osteosarcoma, we identified administration (intraperitoneal) of pectolinarigenin (20 mg/kg/2 days and 50 mg/kg/2 days) blocked STAT3 activation and impaired tumor growth and metastasis with superior pharmacodynamic properties. Taken together, our findings demonstrate that pectolinarigenin may be a candidate for osteosarcoma intervention linked to its STAT3 signaling inhibitory activity.
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Productos Biológicos/farmacología , Cromonas/farmacología , Osteosarcoma/enzimología , Osteosarcoma/patología , Proteína Tirosina Fosfatasa no Receptora Tipo 6/metabolismo , Factor de Transcripción STAT3/metabolismo , Transducción de Señal/efectos de los fármacos , Apoptosis/efectos de los fármacos , Productos Biológicos/toxicidad , Adhesión Celular/efectos de los fármacos , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Cromonas/toxicidad , Transición Epitelial-Mesenquimal/efectos de los fármacos , Humanos , Invasividad Neoplásica , Fenotipo , Fosforilación/efectos de los fármacos , Fosfotirosina/metabolismo , Análisis de Supervivencia , Ensayo de Tumor de Célula Madre , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
CD151, a tetraspanin family protein involved in cell-cell and cell-extracellular matrix interaction, is differentially expressed in osteosarcoma cell membranes. Thus, this study aimed to investigate the role of CD151 in osteosarcoma metastasis. We analyzed CD151 expression in patient tissue samples using immunohistochemistry. CD151 expression was also silenced with shRNA in osteosarcoma cells of high metastatic potential, and cell adhesion, migration and invasion were evaluated in vitro and pulmonary metastasis was investigated in vivo. Mediators of cell signaling pathways were also examined following suppression of CD151 expression. Overall survival for patients with low versus high CD151 expression level was 94 vs. 41 months (p=0.0451). CD151 expression in osteosarcoma cells with high metastatic potential was significantly higher than in those with low metastatic potential (p<0.001). shRNA-mediated silencing of CD151 did not influence cell viability or proliferation; however, cell adhesion, migration and invasion were all inhibited (all p<0.001). In mice inoculated with shRNA-transduced osteosarcoma cells, the number and size of lung metastatic lesions were reduced compared to the mice inoculated with control-shRNA transduced cells (p<0.001). In addition, CD151 knockdown significantly reduced Akt, p38, and p65 phosphorylation as well as focal adhesion kinase, integrin ß1, p70s6, and p-mTOR levels. Taken together, CD151 induced osteosarcoma metastasis likely by regulating cell function through adhesion signaling. Further studies are necessary to fully explore the diagnostic and prognostic value of determining CD151 expression in osteosarcoma patients.