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1.
World Neurosurg ; 2023 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-37343670

RESUMO

OBJECTIVE: To identify the risk factors for developing postoperative pulmonary infection in patients with acute cervical spinal cord injury (CSCI), and to develop a nomogram prediction model. METHODS: Patients with CSCI who were admitted to 3 different medical centers between July 2011 and July 2021 were included in this study. All patients underwent cervical spine surgery. Data for patients admitted to the first 2 centers were included in a training set to establish the nomogram prediction model, and data for patients admitted to the third center were included in a validation set to externally verify the efficacy of the prediction model. For the training set, patients were divided into an infected group and a noninfected group (control group). Independent risk factors for postoperative pulmonary infection in patients with CSCI were identified by univariate and multivariate logistic regression analyses. Additionally, a nomogram prediction model was developed and validated based on the risk factors. RESULTS: A total of 689 patients were enrolled, including 574 for the training set and 115 for the validation set. Of the patients included for the training set, 144 developed pulmonary infection, with an incidence of 25.09%; 40 patients included for the validation set developed pulmonary infection (34.78%). Multivariate logistic regression analysis showed that age, American Spinal Injury Association grade, steroid pulse, high-level injury, smoking, multistage surgery, and operation duration were risk factors for the development of postoperative pulmonary infection in patients with CSCI. The area under the curve of the receiver operating characteristic curve of the model built by the training set was 0.905, and that of the receiver operating characteristic curve of the verification set was 0.917. The decision curve indicated that the model was in the range 1%-100%, and the predicted net benefit value of the model was high. CONCLUSIONS: Age, American Spinal Injury Association grade, steroid pulse, CSCI site, smoking history, number of surgical levels, and surgical duration are correlated with the development of postoperative pulmonary infection in patients with CSCI. The risk prediction model of postoperative pulmonary infection has a good prediction efficiency and accuracy.

2.
Spinal Cord ; 61(6): 323-329, 2023 06.
Artigo em Inglês | MEDLINE | ID: mdl-36894765

RESUMO

STUDY DESIGN: A retrospective study. OBJECTIVE: Traumatic cervical spinal cord injury (TSCI) is often associated with disc rupture. It was reported that high signal of disc and anterior longitudinal ligament (ALL) rupture on magnetic resonance imaging (MRI) were the typical signs of ruptured disc. However, for TSCI with no fracture or dislocation, there is still difficult to diagnose disc rupture. The purpose of this study was to investigate the diagnostic efficiency and localization method of different MRI features for cervical disc rupture in patient with TSCI but no any signs of fracture or dislocation. SETTING: Affiliated hospital of University in Nanchang, China. METHODS: Patients who had TSCI and underwent anterior cervical surgery between June 2016 and December 2021 in our hospital were included. All patients received X-ray, CT scan, and MRI examinations before surgery. MRI findings such as prevertebral hematoma, high-signal SCI, high-signal posterior ligamentous complex (PLC), were recorded. The correlation between preoperative MRI features and intraoperative findings was analyzed. Also, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of these MRI features in diagnosing the disc rupture were calculated. RESULTS: A total of 140 consecutive patients, 120 males and 20 females with an average age of 53 years were included in this study. Of these patients, 98 (134 cervical discs) were intraoperatively confirmed with cervical disc rupture, but 59.1% (58 patients) of them had no definite evidence of an injured disc on preoperative MRI (high-signal disc or ALL rupture signal). For these patients, the high-signal PLC on preoperative MRI had the highest diagnostic rate for disc rupture based on intraoperative findings, with a sensitivity of 97%, specificity of 72%, PPV of 84% and NPV of 93%. Combined high-signal SCI with high-signal PLC had higher specificity (97%) and PPV (98%), and a lower FPR (3%) and FNR (9%) for the diagnosis of disc rupture. And combination of three MRI features (prevertebral hematoma, high-signal SCI and PLC) had the highest accuracy in diagnosing traumatic disc rupture. For the localization of the ruptured disc, the level of the high-signal SCI had the highest consistency with the segment of the ruptured disc. CONCLUSION: MRI features, such as prevertebral hematoma, high-signal SCI and PLC, demonstrated high sensitivities for diagnosing cervical disc rupture. High-signal SCI on preoperative MRI could be used to locate the segment of ruptured disc.


Assuntos
Medula Cervical , Fraturas Ósseas , Luxações Articulares , Traumatismos da Medula Espinal , Traumatismos da Coluna Vertebral , Masculino , Feminino , Humanos , Pessoa de Meia-Idade , Traumatismos da Medula Espinal/complicações , Traumatismos da Medula Espinal/diagnóstico por imagem , Traumatismos da Medula Espinal/cirurgia , Estudos Retrospectivos , Medula Cervical/lesões , Imageamento por Ressonância Magnética , Fraturas Ósseas/complicações , Vértebras Cervicais/diagnóstico por imagem , Vértebras Cervicais/cirurgia , Vértebras Cervicais/lesões
3.
Orthop Surg ; 14(6): 1203-1216, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35470579

RESUMO

OBJECTIVE: To investigate the changes in proinflammatory cytokines and chemokines, namely, C-C motif ligand (CCL) 2 and CCL7, in postmenopausal osteoporosis (PMOP) and to develop a new drug, bindarit (Bnd), for PMOP in an ovariectomized (OVX) mouse model. METHODS: Bone marrow macrophages (BMMs) from the femurs of five women with PMOP and five premenopausal women without osteoporosis were detected by RNA sequencing. BMMs from mice were differentiated into osteoclasts and treated with a synthetic inhibitor of CCL2 and CCL7, Bnd, or 17 beta estradiol (E2 ). Mouse BMMs were differentiated into osteoclasts with or without Bnd for 7 days and analyzed by RNA sequencing. Osteoblasts of mice were induced to undergo osteoblastogenesis and treated with Bnd. OVX mice were treated with E2 or Bnd after surgery. The protein and mRNA expression of CCL2 and CCL7 was detected using immunostaining and qPCR, respectively, in OVX and aged mice and in cells cultured in vitro. Osteoclast formation was detected using a tartrate-resistant acid phosphatase (TRAP) assay in vitro and in vivo. Alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2) and osteocalcin (OCN) were detected using immunostaining to evaluate osteogenesis. Microcomputed tomography was conducted to analyze trabecular bone parameters, the structure model index, bone mineral density and other variables. Nuclear factor-κB (NF-κB) signaling pathway-related protein phosphorylation of IKKα/ß (p-IKKα/ß) and p-NFκB p65 was examined using western blotting. RESULTS: CCL2, CCL7 and their receptor of C-C chemokine receptor-2 (CCR2), and the NF-κB signaling pathway, were significantly increased in women with PMOP. CCL2 and CCL7 protein and mRNA expression was increased in OVX mice and aged female mice, but the increases were attenuated by E2 and Bnd. E2 and Bnd effectively inhibited osteoclastogenesis and the protein expression of CCL2 and CCL7 both in vitro and in vivo and reduced bone loss in OVX mice. Bnd did not affect the mineralization of osteoblasts directly in vitro but reduced bone turnover in vivo. p-IKKα/ß and p-NFκB p65 levels were increased in BMMs of mice after differentiation into osteoclasts but were significantly decreased by Bnd. CONCLUSION: The proinflammatory cytokines and chemokines CCL2, CCL7 and CCR2 were correlated with PMOP. Bnd attenuated the increases in CCL2 and CCL7 levels to affect osteoporosis in OVX mice via the NFκB signaling pathway. Thus, Bnd may be useful as a new therapeutic for the prevention of PMOP.


Assuntos
Doenças Ósseas Metabólicas , Reabsorção Óssea , Osteoporose Pós-Menopausa , Osteoporose , Animais , Diferenciação Celular , Quimiocina CCL2 , Quimiocina CCL7 , Citocinas/metabolismo , Feminino , Humanos , Quinase I-kappa B/metabolismo , Quinase I-kappa B/farmacologia , Indazóis , Camundongos , NF-kappa B/metabolismo , Osteoclastos , Osteogênese , Osteoporose/tratamento farmacológico , Osteoporose/metabolismo , Osteoporose Pós-Menopausa/metabolismo , Ovariectomia , Propionatos , RNA Mensageiro/metabolismo , Transdução de Sinais , Microtomografia por Raio-X
4.
World Neurosurg ; 123: e371-e378, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30500586

RESUMO

OBJECTIVE: To retrospectively analyze the outcomes and complications of patients with metastatic thoracic spinal tumors (MTTs) who underwent posterior corpectomies. METHODS: Ninety patients with MTTs who underwent posterior corpectomies were retrospectively analyzed. Characteristics evaluated included number of MTTs per year, location, involved vertebrae numbers, sex, histology, pre- and postoperative American Spinal Injury Association (ASIA) grade, visual analog scale (VAS) pain scores, operative time, blood loss, and length of hospital stay. RESULTS: The average follow-up was 20.8 ± 27.9 months (range, 0.5-139.4 months). Of the patients, 76.67% had a single metastasis and 23.33% had multiple metastases. For histology, 16.67% were breast, 15.56% were lung, 12.22% were prostate, and 12.22% were renal cell carcinoma. Of the patients with paraplegia and paraparesis, 74% improved. One patient improved from ASIA grade A to D, 3 patients improved from grade B to C, 8 patients improved from grade C to D or E, and 25 patients improved from grade D to E. Three patients (6%) with ASIA grade A and 1 patient (2%) with ASIA grade B had no improvement. One patient with ASIA grade C and 8 patients (16%) with grade D had no improvement. After surgery, VAS pain scores decreased from 8.45 ± 1.57 to 1.211 ± 1.81. In terms of complications, 2 patients (2.22%) had deep vein thrombosis and 1 patient had pulmonary embolism (1.11%). Other complications included wound infection (4.44%), cerebrospinal fluid leak (4.44%), pleural effusion (3.33%), wound dehiscence (2.22%), cellulitis (1.11%), epidural hematoma (1.11%), and pneumothorax (1.11%). Of the patients, 2.22% had implant failure and pseudoarthrosis, with 1 patient needing revision surgery. One patient (1.11%) had tumor recurrence. CONCLUSIONS: Our results suggest that posterior thoracic corpectomies for MTTs have a reasonable complication rate with favorable outcomes.


Assuntos
Neoplasias da Coluna Vertebral/cirurgia , Vértebras Torácicas/cirurgia , Adulto , Idoso , Idoso de 80 Anos ou mais , Perda Sanguínea Cirúrgica/estatística & dados numéricos , Descompressão Cirúrgica/métodos , Feminino , Humanos , Tempo de Internação/estatística & dados numéricos , Masculino , Metastasectomia/métodos , Pessoa de Meia-Idade , Duração da Cirurgia , Dor Pós-Operatória , Paraparesia/etiologia , Paraparesia/cirurgia , Paraplegia/etiologia , Paraplegia/cirurgia , Complicações Pós-Operatórias/etiologia , Estudos Retrospectivos , Neoplasias da Coluna Vertebral/secundário , Resultado do Tratamento , Adulto Jovem
5.
PLoS One ; 11(12): e0166751, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-28033335

RESUMO

Cortex Eucommiae is used worldwide in traditional medicine, various constituents of Cortex Eucommiae, such as chlorogenic acid (CGA), has been reported to exert anti-osteoporosis activity in China, but the mechanism about their contribution to the overall activity is limited. The aims of this study were to determine whether chlorogenic acid can prevent estrogen deficiency-induced osteoporosis and to analyze the mechanism of CGA bioactivity. The effect of CGA on estrogen deficiency-induced osteoporosis was performed in vivo. Sixty female Sprague-Dawley rats were divided randomly among a sham-operated group and five ovariectomy (OVX) plus treatment subgroups: saline vehicle, 17α-ethinylestradiol (E2), or CGA at 9, 27, or 45 mg/kg/d. The rats' femoral metaphyses were evaluated by micro-computed tomography (µCT). The mechanism of CGA bioactivity was investigated in vitro. Bone mesenchymal stem cells (BMSCs) were treated with CGA, with or without phosphoinositide 3-kinase (PI3K) inhibitor LY294002. BMSCs proliferation and osteoblast differentiation were assessed with 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and alkaline phosphatase, with or without Shp2 interfering RNA (RNAi). The results display that CGA at 27 and 45 mg/kg/day inhibited the decrease of bone mineral density (BMD) that induced by OVX in femur (p< 0.01), significantly promoted the levels of bone turnover markers, and prevented bone volume fraction (BV/TV), connectivity density (CoonD), trabecular number (Tb.N), trabecular thickness (Tb.Th) (all p< 0.01) to decrease and prevented the trabecular separation (Tb.Sp), structure model index (SMI)(both p< 0.01) to increase. CGA at 1 or 10 µM enhanced BMSC proliferation in a dose-dependent manner. CGA at 0.1 to 10 µM increased phosphorylated Akt (p-Akt) and cyclin D1. These effects were reversed by LY294002. CGA at 1 or 10 µM increased BMSC differentiation to osteoblasts (p< 0.01), Shp2 RNAi suppressed CGA-induced osteoblast differentiation by decreasing Shp2, p-Akt, and cyclin D1. This study found that CGA improved the BMD and trabecular micro-architecture for the OVX-induced osteoporosis. Therefore, CGA might be an effective alternative treatment for postmenopausal osteoporosis. CGA promoted proliferation of osteoblast precursors and osteoblastic differentiation of BMSCs via the Shp2/PI3K/Akt/cyclin D1 pathway.


Assuntos
Densidade Óssea/efeitos dos fármacos , Ácido Clorogênico/farmacologia , Osteoblastos/citologia , Osteogênese/efeitos dos fármacos , Osteoporose Pós-Menopausa/prevenção & controle , Animais , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Cromonas/farmacologia , Ciclina D1/metabolismo , Feminino , Humanos , Células-Tronco Mesenquimais/citologia , Morfolinas/farmacologia , Osteoblastos/metabolismo , Ovariectomia , Fosfatidilinositol 3-Quinases/metabolismo , Inibidores de Fosfoinositídeo-3 Quinase , Proteína Tirosina Fosfatase não Receptora Tipo 11/genética , Proteína Tirosina Fosfatase não Receptora Tipo 11/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Interferência de RNA , RNA Interferente Pequeno/genética , Ratos , Ratos Sprague-Dawley
6.
Mol Med Rep ; 11(6): 4489-95, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25634525

RESUMO

Chlorogenic acid (CGA) exhibits various biological properties, including the inhibition of oxidation, obesity, apoptosis and tumorigenesis. CGA is also able to promote cell survival and proliferation. The aim of the present study was to determine the effects and underlying molecular mechanisms of CGA on the adipogenesis of bone marrow­derived mesenchymal stem cells (BMSCs). Treatment with CGA had a marginal effect on cell proliferation, by promoting the expression levels of phosphorylated Akt and cyclin D1. Furthermore, treatment with CGA also upregulated the phosphorylation of extracellular signal­regulated kinase (Erk) and inhibited the adipocyte differentiation of BMSCs by inhibiting the expression of peroxisome proliferator­activated receptor (PPAR)γ and CCAAT/enhancer binding protein (C/EBP)α. However, knockdown of the expression of Shp2 attenuated CGA­induced proliferation and inhibited the phosphorylation of Akt and expression of cyclin D1. Furthermore, CGA treatment upregulated Erk phosphorylation and decreased the expression levels of PPARγ and CEBPα, which was inhibited by treatment with the Shp2 PTPase activity inhibitor, NSC­87877. The results of the present study suggested that CGA­induced Akt and Erk pathways regulate proliferation and differentiation and that Shp2 is important in the proliferation and differentiation of BMSCs.


Assuntos
Adipogenia/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Ácido Clorogênico/farmacologia , Células-Tronco Mesenquimais/metabolismo , Proteína Tirosina Fosfatase não Receptora Tipo 11/metabolismo , Adulto , Células da Medula Óssea/citologia , Proteínas Estimuladoras de Ligação a CCAAT/metabolismo , Ciclina D1/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Feminino , Humanos , Masculino , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , PPAR gama/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação/efeitos dos fármacos , Proteína Tirosina Fosfatase não Receptora Tipo 11/antagonistas & inibidores , Proteína Tirosina Fosfatase não Receptora Tipo 11/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Quinolinas/farmacologia , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Transdução de Sinais/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacos
7.
Mol Med Rep ; 11(2): 1352-7, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25351625

RESUMO

Increasing evidence suggests that fatty acid synthase (FASN) is crucial in the carcinogenesis of various types of tumor. In addition, the phosphatidylinositol 3­kinase (PI3K)/Akt signaling pathway, which is closely associated with cellular metabolism, affects cancer biology. However, whether the malignant phenotype of osteosarcoma (OS) cells is regulated by the PI3K/Akt/FASN signaling pathway and how the PI3K family specific inhibitor, 2­(4­morpholinyl)­8­phenyl­chromone (LY294002) affects the malignant phenotype of OS cells remains to be elucidated. In the present study, U2­OS and MG­63 cells were treated with LY294002 and subsequently western blot analysis was used to examine Akt, p­Akt and FASN protein expression. Additionally, FASN mRNA was detected by reverse transcription quantitative polymerase chain reaction. MTT and fluorescence­activated cell sorting assays were used to assess proliferation and apoptosis. Migration and invasion were investigated using wound healing and transwell invasion assays. The results demonstrated that LY294002 suppressed the PI3K/Akt/FASN signaling pathway. However, the malignant phenotypes of OS cells mentioned above were significantly inhibited. The present results indicated that LY294002 inhibits the malignant phenotype of OS cells via modulation of the PI3K/Akt/FASN signaling pathway in vitro and may be a new therapeutic strategy for the management of OS.


Assuntos
Cromonas/farmacologia , Ácido Graxo Sintases/metabolismo , Morfolinas/farmacologia , Fosfatidilinositol 3-Quinase/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/efeitos dos fármacos , Neoplasias Ósseas/metabolismo , Neoplasias Ósseas/patologia , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Ácido Graxo Sintases/genética , Humanos , Osteossarcoma/metabolismo , Osteossarcoma/patologia , Fenótipo , RNA Mensageiro/metabolismo
8.
Int J Mol Sci ; 15(10): 18525-39, 2014 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-25318055

RESUMO

Chalcone derivatives (E)-3-(4-hydroxy-3-methoxyphenyl)-1-(4-methoxyphenyl) prop-2-en-1-one and (E)-3-(4-hydroxyphenyl)-1-(4-methoxyphenyl) prop-2-en-1-one (Compounds 1 and 2) have been demonstrated to be potent anti-inflammatory agents in our previous study. In light of the relationship of intracellular mechanisms between anti-inflammatories and antioxidants, we further designed and synthesized a series of chalcone derivatives based on 1 and 2, to explore their antioxidant efficacy. The majority of the derivatives exhibited strong protective effects on PC12 (PC12 rat pheochromocytoma) cells exposed to H2O2, and all compounds were nontoxic. A preliminary structure-activity relationship was proposed. Compounds 1 and 1d ((E)-2-methoxy-4-(3-(4-methoxyphenyl)-3-oxoprop-1-en-1-yl) phenyl acrylate) exerted the action in a good dose-dependent manner. Quantitative RT-PCR (qRT-PCR) and western blot analysis showed that 1 and 1d significantly improve the expression of nuclear factor erythroid 2 p45-related factor 2 (Nrf2)-dependent antioxidant genes g-Glutamylcysteine Ligase Catalytic Subunit (GCLC) and heme oxygenase-1 (HO-1) and their corresponding proteins (γ-glutamyl cysteine synthase (γ-GCS) and HO-1) in PC12 cells. Inhibition of GCLC and HO-1 by specific inhibitors, L-buthionine-S-sulfoximine (BSO) and zinc protoporphyrin (ZnPP), respectively, partially reduce the protective effect of 1 and 1d. These data present a series of novel chalcone analogs, especially compounds 1 and 1d, as candidates for treating oxidative stress-related disease by activating the Nrf2-antioxidant responsive element (ARE) pathway.


Assuntos
Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Chalconas/farmacologia , Neurônios/citologia , Neurônios/efeitos dos fármacos , Animais , Antioxidantes/química , Chalconas/química , Peróxido de Hidrogênio/metabolismo , Neurônios/metabolismo , Células PC12 , Ratos
9.
Asian Pac J Cancer Prev ; 15(3): 1241-5, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24606447

RESUMO

Cinobufacin is used clinically to treat patients with many solid malignant tumors. However, the mechanisms underlying action remain to be detailed. Our study focused on miRNAs involved in cinobufacin inhibition of GC cell proliferation. miRNA microarray analysis and real time PCR identified miR-494 as a significant cinobufacin- associated miRNA. In vivo, ectopic expression of miR-494 inhibited the proliferation and induced apoptosis of BGC-823 cells on CCK-8 and flow cytometry analysis. Further study verified BAG-1 (anti-apoptosis gene) to bea target of miR-494 by luciferase reporter assay and Western blotting. In summary, our study demonstrated that cinobufacin may inhibit the proliferation and promote the apoptosis of BGC-823 cells. Cinobufacin-associated miR-494 may indirectly be involved in cell proliferation and apoptosis by targeting BAG-1, pointing to use as a potential molecular target of cinobufacin in gastric cancer therapy.


Assuntos
Venenos de Anfíbios/farmacologia , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Proteínas de Ligação a DNA/biossíntese , MicroRNAs/biossíntese , Fatores de Transcrição/biossíntese , Regiões 3' não Traduzidas/genética , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Proteínas de Ligação a DNA/genética , Humanos , Medicina Tradicional Chinesa , MicroRNAs/genética , Sincalida , Neoplasias Gástricas/tratamento farmacológico , Fatores de Transcrição/genética , Transfecção
10.
Int J Mol Med ; 33(3): 633-9, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24366211

RESUMO

The activation of PI3K/Akt and the overexpression of fatty acid synthase (FASN) are frequently observed in human osteosarcoma (OS). In the present study, in order to investigate the possible association between the phosphorylation of Akt and FASN expression, immunohistochemical staining was conducted on 24 OS specimens from patients with pulmonary metastasis, which revealed a significant positive correlation between phosphorylated Akt (p-Akt) and the expression of FASN (R=0.469, P=0.04). To investigate the association between p-Akt and FASN in vitro, human U2-OS OS cells were treated with FASN-specific RNAi plasmid or LY294002 (an inhibitor of PI3k/Akt). The mRNA levels of Akt and FASN were measured by real-time PCR. Western blot analysis was also performed to detect the protein experession of PI3K, Akt, p-Akt and FASN. The results demonstrated that the PI3K/Akt signaling pathway modulates FASN expression; the inhibition of FASN resulted in the downregulation of p-Akt in the U2-OS cells. Furthermore, the effects induced by the inhibition of the activity of p-Akt or FASN on the malignant phenotype of U2-OS cells were investigated, demonstrating that the malignant phenotype was inhibited by suppressing the activity of PI3K/Akt or FASN in the U2-OS cells. The findings from our study suggest the existence of a positive feedback regulation between Akt phosphorylation and FASN expression and that this loop may play an important role in the malignant phenotype of OS cells.


Assuntos
Ácido Graxo Sintase Tipo I/metabolismo , Osteossarcoma/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/genética , Apoptose/genética , Proliferação de Células , Ácido Graxo Sintase Tipo I/biossíntese , Retroalimentação Fisiológica , Regulação Neoplásica da Expressão Gênica , Humanos , Osteossarcoma/genética , Osteossarcoma/patologia , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação , Proteínas Proto-Oncogênicas c-akt/biossíntese
11.
Oncol Rep ; 31(1): 328-34, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24172910

RESUMO

Lapatinib, an inhibitor of human epidermal growth factor receptor 2 (HER2) phosphorylation, has been reported to inhibit several types of tumors such as HER2-overexpressing breast cancer. However, the effect of lapatinib on the malignant phenotype of human osteosarcoma (OS) cells and the potential molecular mechanisms remain unclear. To elucidate the effect of lapatinib on OS, two OS cell lines, U2-OS and MG-63, were utilized in the present study. Various concentrations of lapatinib were used to treat OS cells for different time durations. Cell proliferation was evaluated by MTT and colony formation assays. Flow cytometry (FCM) was used to evaluate cell apoptosis. Wound healing and Transwell invasion assays were performed to examine the migratory and invasive abilities of the cells. To investigate the possible molecular mechanisms involved, the expression of p-HER2, phosphatidylinositol 3-kinase (PI3K), p-AKT, AKT and fatty acid synthase (FASN) protein was detected by western blotting. MTT assays showed that lapatinib inhibited the proliferation of U2-OS and MG-63 cells in a dose- and time-dependent manner, and the rate of colony formation of the lapatinib-treated cells was significantly lower when compared to those cells not treated with lapatinib in both cell lines. FCM assay revealed a significantly higher apoptotic rate in the lapatinib-treated OS cells. Wound healing and Transwell invasion assays revealed that the migratory and invasive abilities of OS cells were significantly inhibited by lapatinib (P<0.05). Western blotting showed that lapatinib suppressed the activity of HER2-PI3K/AKT-FASN in U2-OS and MG-63 cells in vitro. These results suggest that lapatinib may alter the malignant phenotype of OS cells via downregulation of the activity of the HER2-PI3K/AKT-FASN signaling pathway in vitro. Thus, lapatinib may be an effective chemotherapeutic agent for the treatment of osteosarcoma.


Assuntos
Antineoplásicos/farmacologia , Neoplasias Ósseas/patologia , Osteossarcoma/patologia , Inibidores de Proteínas Quinases/farmacologia , Quinazolinas/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias Ósseas/metabolismo , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Ácido Graxo Sintase Tipo I/biossíntese , Humanos , Lapatinib , Invasividade Neoplásica , Osteossarcoma/metabolismo , Fosfatidilinositol 3-Quinases/biossíntese , Proteínas Proto-Oncogênicas c-akt/biossíntese , Receptor ErbB-2/antagonistas & inibidores , Receptor ErbB-2/biossíntese , Transdução de Sinais/efeitos dos fármacos
12.
Int J Clin Exp Pathol ; 7(12): 8777-83, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25674245

RESUMO

Elevated expression of fatty acid synthase (FASN) and human epidermal growth factor receptor-2 (HER2) are observed in human osteosarcoma (OS). The aim in this study is to investigate a possible connection between FASN expression and the activity of HER2. The immunohistochemistry staining was conducted on 24 OS specimens from patients, which revealed a significant positive correlation between FASN and HER2 as well as p-HER2 protein expression. Furthermore, the human OS cell lines MG-63 and U2-OS were treated with FASN-specific RNAi Plasmid or Lapatinib (an inhibitor of HER2). The mRNA of HER2 and FASN was measured using RT-PCR. Western blot was performed to detect the protein expression of HER2, p-HER2 and FASN. The results demonstrated that HER2 modulates FASN expression, inhibition of FASN resulted in down-regulation of HER2 and p-HER2 protein in OS cells. Our findings suggested that there was positive feedback regulation between FASN and HER2 expression and phosphorylation in OS cells.


Assuntos
Ácido Graxo Sintase Tipo I/metabolismo , Regulação Neoplásica da Expressão Gênica/fisiologia , Osteossarcoma/metabolismo , Receptor ErbB-2/metabolismo , Western Blotting , Humanos , Imuno-Histoquímica , Reação em Cadeia da Polimerase em Tempo Real
13.
Biochem Biophys Res Commun ; 440(2): 229-34, 2013 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-24041695

RESUMO

FASN plays an important role in the malignant phenotype of various tumors. Our previous studies show that inhibition FASN could induce apoptosis and inhibit proliferation in human osteosarcoma (OS) cell in vivo and vitro. The aim in this study was to investigate the effect of inhibition FASN on the activity of HER2/PI3K/AKT axis and invasion and migration of OS cell. The expression of FASN, HER2 and p-HER2(Y1248) proteins was detected by immunohistochemistry in OS tissues from 24 patients with pulmonary metastatic disease, and the relationship between FASN and p-HER2 as well as HER2 was investigated. The results showed that there was a positive correlation between FASN and HER2 as well as p-HER2 protein expression. The U-2 OS cells were transfected with either the FASN specific RNAi plasmid or the negative control RNAi plasmid. FASN mRNA was measured by RT-PCR. Western blot assays was performed to examine the protein expression of FASN, HER2, p-HER2(Y1248), PI3K, Akt and p-Akt (Ser473). Migration and invasion of cells were investigated by wound healing and transwell invasion assays. The results showed that the activity of HER2/PI3K/AKT signaling pathway was suppressed by inhibiting FASN. Meanwhile, the U-2OS cells migration and invasion were also impaired by inhibiting the activity of FASN/HER2/PI3K/AKT. Our results indicated that inhibition of FASN suppresses OS cell invasion and migration via down-regulation of the "HER2/PI3K/AKT" axis in vitro. FASN blocker may be a new therapeutic strategy in OS management.


Assuntos
Ácido Graxo Sintases/antagonistas & inibidores , Osteossarcoma/tratamento farmacológico , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Regulação para Baixo , Humanos , Invasividade Neoplásica/prevenção & controle , Fosfatidilinositol 3-Quinases , Proteínas Proto-Oncogênicas c-akt , Interferência de RNA , Receptor ErbB-2/antagonistas & inibidores , Transdução de Sinais/efeitos dos fármacos
14.
Asian Pac J Cancer Prev ; 14(6): 3871-5, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23886199

RESUMO

PURPOSE: MicroRNAs (miRNAs) are small endogenous, non-coding, single-stranded RNAs (approximately 22 nt). Accumulating evidence has shown that aberrant miRNA expression is pronounced and correlated with gastric cancer genesis and progression. MATERIALS AND METHODS: Expression levels of miR-181a-5p in GC tissues and cell lines were assessed by qRT-PCR and tested for correlation with clinical features. In addition, effects of miR-181a-5p on GC cell growth were investigated. RESULTS: Our findings indicate that miR-181a-5p is upregulated in GC, in correlation with lymph node invasion, nerve invasion and vascular invasion (P<0.05). Enforced expression of miR-181a -5p promoted cell proliferation ability. CONCLUSIONS: This study suggested that increased miR-181a-5p is related to GC progression. MiR-181a-5p may represent a potential therapeutic target for GC.


Assuntos
Adenoma/patologia , Biomarcadores Tumorais/genética , Proliferação de Células , Mucosa Gástrica/metabolismo , MicroRNAs/genética , Neoplasias Gástricas/patologia , Adenoma/genética , Células Cultivadas , Perfilação da Expressão Gênica , Humanos , Metástase Linfática , Invasividade Neoplásica , Estadiamento de Neoplasias , Análise de Sequência com Séries de Oligonucleotídeos , Prognóstico , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estômago/patologia , Neoplasias Gástricas/genética
15.
Indian J Orthop ; 47(6): 553-8, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24379459

RESUMO

BACKGROUND: Anterior cervical interbody grafts/cages combined with a plate were frequently used in multilevel discectomies/corpectomies. In order to avoid additional posterior stabilization in patients who undergo anterior reconstructive surgery, an anterior cervical transpedicular screw fixation, which offers higher stability is desirable. We investigated in this study the anatomical (morphologic) characters for cervical anterior transpedicular screw fixation. MATERIALS AND METHODS: Left pedicle parameters were measured on computed tomography (CT) images based on 36 cervical spine CT scans from healthy subjects. The parameters included outer pedicle width (Distance from lateral to medial pedicle surface in the coronal plane), outer pedicle height (OPH) (Distance from upper to lower pedicle surface in the sagittal plane), maximal pedicle axis length (MPAL), distance transverse insertion point (DIP), distance of the insertion point to the upper end plate (DIUP), pedicle sagittal transverse angle (PSTA) and pedicle transverse angle (PTA) at C3 to C7. RESULTS: The values of outer pedicle width and MPAL in males were larger than in females from C3 to C7. The OPH in males was larger than in females at C3 to C6, but there was no difference at C7. The DIP and PTA were significantly greater in males than in females at C3, but there was no difference in the angle at C4-7. The PSTA was not statistically different between genders at C3, 4, 7, but this value in males was larger than females at C5, 6. The DIUP was significantly greater in males at C3, 4, 6, 7 but was non significant at C5. CONCLUSIONS: The placement of cervical anterior transpedicular screws should be individualized for each patient and based on a detailed preoperative planning.

16.
Oncol Lett ; 4(5): 1125-1129, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23162665

RESUMO

microRNAs are involved in different cancer-related processes. miR-195, one of the miR-16/15/195/424/497 family members, has been shown to act as a tumor suppressor during tumorigenesis. However, the function of miR-195 in osteosarcoma is still unclear. In our study, the miR-195 expression level was upregulated in osteosarcoma cells, by transfection with miR-195, and the fatty acid synthase (FASN) mRNA and protein expression levels were measured by RT-PCR and western blotting. Cell migration and invasion was measured using wound healing migration and Transwell invasion assays. We found that the upregulation of miR-195 greatly decreased cell invasion and the migration of U2OS. We also identified that FASN may be a direct target of miR-195 by the luciferase activity assay. These findings provide evidence that miR-195 plays a key role in inhibiting osteosarcoma cell migration and invasion through targeting FASN, and strongly suggest that exogenous miR-195 may have therapeutic value in treating osteosarcoma.

17.
J Cell Biochem ; 112(10): 2902-9, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21618594

RESUMO

Anti-diabetic drug metformin has been shown to enhance osteoblasts differentiation and inhibit osteoclast differentiation in vitro and prevent bone loss in ovariectomized (OVX) rats. But the mechanisms through which metformin regulates osteoclastogensis are not known. Osteoprotegerin (OPG) and receptor activator of nuclear factor κB ligand (RANKL) are cytokines predominantly secreted by osteoblasts and play critical roles in the differentiation and function of osteoclasts. In this study, we demonstrated that metformin dose-dependently stimulated OPG and reduced RANKL mRNA and protein expression in mouse calvarial osteoblasts and osteoblastic cell line MC3T3-E1. Inhibition of AMP-activated protein kinase (AMPK) and CaM kinase kinase (CaMKK), two targets of metformin, suppressed endogenous and metformin-induced OPG secretion in osteoblasts. Moreover, supernatant of osteoblasts treated with metformin reduced formation of tartrate resistant acid phosphatase (TRAP)-positive multi-nucleated cells in Raw264.7 cells. Most importantly, metformin significantly increased total body bone mineral density, prevented bone loss and decreased TRAP-positive cells in OVX rats proximal tibiae, accompanied with an increase of OPG and decrease of RANKL expression. These in vivo and in vitro studies suggest that metformin reduces RANKL and stimulates OPG expression in osteoblasts, further inhibits osteoclast differentiation and prevents bone loss in OVX rats.


Assuntos
Metformina/farmacologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Osteoprotegerina/metabolismo , Ligante RANK/metabolismo , Animais , Western Blotting , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Ensaio de Imunoadsorção Enzimática , Feminino , Camundongos , Osteoporose/prevenção & controle , Ovariectomia , Ligante RANK/genética , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais/efeitos dos fármacos , Crânio/citologia , Tíbia/metabolismo
18.
Nan Fang Yi Ke Da Xue Xue Bao ; 30(11): 2436-9, 2010 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-21097398

RESUMO

OBJECTIVE: To observe the effects of lithium chloride combined with human umbilical cord blood mesenchymal stem cell (hUCB-SCs) transplantation in the treatment of spinal cord injury in rats. METHODS: Eighty female SD rats with complete T9 spinal cord transaction were randomized into 4 groups (n=20), namely the control group (group A), lithium chloride group (group B), hUCB-SCs group (group C) and hUCB-SCs(+) lithium chloride group (group D). On days 1 and 3 and the last days of the following weeks postoperatively, the motor function of the hindlimb of the rats were evaluated according to the BBB scores. At 8 weeks, all the rats were sacrificed and the spinal cords were taken for morphological observation. The spinal cord tissues at the injury site were observed with Brdu nuclear labeling to identify the survival and migration of the transplanted SCs. The regeneration and distribution of the spinal nerve fibers were observed with fluorescent-gold (FG) spinal cord retrograde tracing. RESULTS: Brdu labeling showed that the transplanted hUCB-SCs survived and migrated in the spinal cord 8 weeks postoperatively in groups C and D. FG retrograde tracing identified a small amount of pyramidal cells that migrated across the injury site in groups C and D. The BBB scores of the hindlimb motor function 8 weeks postoperatively were 4.11∓0.14, 4.50∓0.15, 8.31∓0.11 and 11.15∓0.18 in groups A, B, C and D, respectively. CONCLUSION: Lithium chloride can promote the survival and differentiation of hUCB-SCs into neural cells at the injury site. Lithium chloride combined with hUCB-SCs transplantation may accelerate functional recovery of the hindlimbs in rats with complete transection of the spinal cord.


Assuntos
Transplante de Células-Tronco de Sangue do Cordão Umbilical , Cloreto de Lítio/uso terapêutico , Traumatismos da Medula Espinal/terapia , Animais , Feminino , Humanos , Ratos
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