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1.
World Neurosurg ; 133: e165-e172, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31476465

RESUMO

BACKGROUND: Cartilaginous endplate (CEP), a thin layer of hyaline cartilage located between the vertebral endplate and nucleus pulposus, transports the nutrient into the disc. The objective of this study was to evaluate the influence of T140 (polyphemusin II-derived peptide) on the CEP cell growth, apoptosis, and the matrix formation via the stromal cell-derived factor-1 (SDF-1)/cysteine X cysteine (CXC) receptor-4 (CXCR4) signaling pathway. METHODS: Sprague-Dawley rats were euthanized by cervical dislocation and dissected for the isolation and the appraisal of CEP cells that were extracted from the endplate in rat intervertebral discs and were then added with different concentrations of reagents (SDF-1 and T140). The effect of T140 on CEP cell proliferation and apoptosis were analyzed. The messenger RNA (mRNA) and protein expressions of CXCR4, prominin-1, proteoglycans, type II collagen, B-cell lymphoma-2 (Bcl-2), and Bcl-2 associated X protein were analyzed by reverse transcription quantitative polymerase chain reaction and Western blot analysis. RESULTS: T140 promoted the proliferation of CEP cells and inhibited the apoptosis of CEP cells. Additionally, T140 suppressed the mRNA and protein expression of CXCR4, prominin-1, and Bcl-2 associated X protein, and increased the mRNA and protein expression of proteoglycans, type II collagen, and Bcl-2. CONCLUSIONS: T140 promotes the proliferation and matrix formation and inhibits the apoptosis of CEP cells by blocking the SDF-1/CXCR4 signaling pathway in vitro, which provides a certain therapeutic effect on the degeneration of intervertebral discs.


Assuntos
Apoptose/efeitos dos fármacos , Quimiocina CXCL12/fisiologia , Condrócitos/efeitos dos fármacos , Matriz Extracelular/efeitos dos fármacos , Cartilagem Hialina/citologia , Disco Intervertebral/citologia , Oligopeptídeos/farmacologia , Receptores CXCR4/fisiologia , Transdução de Sinais/efeitos dos fármacos , Animais , Divisão Celular/efeitos dos fármacos , Proteínas da Matriz Extracelular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Disco Intervertebral/efeitos dos fármacos , Masculino , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley
2.
Mol Med Rep ; 20(2): 1203-1211, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31173219

RESUMO

Low back pain (LBP) is a ubiquitous disease affecting quality of life. The ingrowth of new blood vessels is an important pathological feature of LBP, but its underlying mechanisms are poorly understood. The present study aimed to investigate the influence and relative mechanism of stromal cell derived factor 1 (SDF1) on the angiogenesis of degenerated intervertebral discs. The expression of SDF1 in nucleus pulposus cells (NPCs) was upregulated and downregulated by virus transfection, and the NPCs were allocated to either the downregulation (Down), degeneration (D) or upregulation (Up) group according to the expression of SDF1. The different groups of NPCs or NPC conditioned media were co­cultured with vascular endothelial cells (VECs) under different conditions. A Cell Counting Kit­8 (CCK­8) assay, a Transwell migration assay and a tube formation assay were conducted to evaluate the influence on angiogenesis. The results showed that SDF1 was significantly up­ and downregulated in the Up and Down groups, respectively. Each group of NPCs or their conditioned medium was co­cultured with VECs; the CCK­8, Transwell migration and tube formation assays showed that cell viability, chemotactic migration and the tube formation ability of VECs increased with the rise in SDF1. The aforementioned results were significantly different between each group. After adding the CXCR4 inhibitor, AMD3100, the viability, migration and tube formation of VECs were suppressed in the D and Up groups, and there was a significant difference compared with the prior to the addition of the inhibitor, while there was a declining tendency in the Down group and no significant difference following addition of the inhibitor. The results demonstrated that SDF1 is expressed in human NPCs, and the SDF1/CXCR4 axis can influence the viability, migration and tube formation of VECs and may play an important role in the angiogenesis of human degenerated discs.


Assuntos
Quimiocina CXCL12/metabolismo , Degeneração do Disco Intervertebral/metabolismo , Neovascularização Patológica , Núcleo Pulposo/metabolismo , Transdução de Sinais , Idoso , Células Cultivadas , Quimiocina CXCL12/fisiologia , Técnicas de Cocultura , Feminino , Células Endoteliais da Veia Umbilical Humana , Humanos , Degeneração do Disco Intervertebral/fisiopatologia , Masculino , Pessoa de Meia-Idade , Receptores CXCR4/metabolismo
3.
Biosci Biotechnol Biochem ; 83(6): 1072-1076, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30870116

RESUMO

Diabetes induced a serious of complications including diabetic retinopathy. Our study aimed to investigate the role of Stromal cell-derived factor 1 (SDF-1) and its receptor CXCR4 in diabetic retinopathy. A mice model of diabetic retinopathy was established, and expression of SDF-1 and CXCR4 in retina was examined by Real-time quantitative PCR (qRT-PCR). Cells of human retinal pigment epithelial cell line ARPE-19 were treated with CXCR4 siRNAs and expression vector, and cell viability was detected by MTT assay. We found that expression of SDF-1 and CXCR4 in retina was significantly downregulated in mice with diabetic retinopathy than in normal healthy mice. High glucose treatment downregulated the expression of SDF-1 and CXCR4 in ARPE-19 cells at both mRNA and protein levels. Transfection with CXCR4 siRNAs decreased, while transfection with CXCR4 expression vector increased cell viability under high glucose treatment. We concluded that SDF-1/CXCR4 pathway improved diabetic retinopathy possibly by increasing cell viability. Abbreviations: SDF-1: Stromal cell-derived factor 1; CXCL12: C-X-C motif chemokine 12; qRT-PCR: Real-time quantitative PCR.


Assuntos
Quimiocina CXCL12/fisiologia , Retinopatia Diabética/fisiopatologia , Receptores CXCR4/fisiologia , Animais , Linhagem Celular , Sobrevivência Celular , Quimiocina CXCL12/genética , Retinopatia Diabética/metabolismo , Modelos Animais de Doenças , Regulação para Baixo , Inativação Gênica , Glucose/administração & dosagem , Humanos , Masculino , Camundongos Endogâmicos C57BL , RNA Mensageiro/genética , RNA Interferente Pequeno/genética , Reação em Cadeia da Polimerase em Tempo Real , Receptores CXCR4/genética , Epitélio Pigmentado da Retina/citologia
4.
Biomed Pharmacother ; 112: 108649, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30784930

RESUMO

Titanium surface modification is widely established and has been proven to improve the osseointegration, but the molecular mechanism remains to be fully elucidated. MicroRNAs serve vital roles in the process of regulating osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs). In this study, we report that miR-23a was significantly down-regulated in the osteogenic differentiation process of BMSCs on nanostructured titanium surfaces. Elevated miR-23a inhibited osteogenic differentiation of BMSCs, and decreased miR-23a enhanced this process. In addition, we also observed that CXCL12 was a direct target of miR-23a. Knockdown of CXCL12 inhibited nanotube Ti induced-osteogenic differentiation of BMSCs, similar to the effect of upregulation of miR-23a. Finally, p53 was decreased and it regulated miR-23a/CXCL12 axis during nanotube Ti induced-osteogenic differentiation of BMSCs. Therefore, our findings suggest that by targeting CXCL12, miR-23a serves a vital role in osteogenic differentiation of BMSCs cultured on nanostructured titanium surfaces, which may provide novel clinical treatments for osseointegration.


Assuntos
Quimiocina CXCL12/fisiologia , Células-Tronco Mesenquimais/fisiologia , MicroRNAs/fisiologia , Nanoestruturas , Osteogênese/fisiologia , Proteína Supressora de Tumor p53/fisiologia , Células da Medula Óssea/fisiologia , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/fisiologia , Células Cultivadas , Humanos , Células-Tronco Mesenquimais/efeitos dos fármacos , Nanoestruturas/administração & dosagem , Osteogênese/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia , Propriedades de Superfície/efeitos dos fármacos , Titânio/administração & dosagem
5.
J Orthop Res ; 37(6): 1294-1302, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30345545

RESUMO

A significant number of fractures develop non-union. Mesenchymal stem cell (MSC) therapy may be beneficial, however, this requires cell acquisition, culture and delivery. Endogenous mobilization of stem cells offers a non-invasive alternative. The hypothesis was administration of VEGF and the CXCR4 antagonist AMD3100 would increase the circulating pool of available MSCs and improve fracture healing. Ex-breeder female wistar rats received VEGF followed by AMD3100, or sham PBS. Blood prepared for culture and colonies were counted. P3 cells were analyzed by flow cytometry, bi-differentiation. The effect of mobilization on fracture healing was evaluated with 1.5 mm femoral osteotomy stabilized with an external fixator in 12-14 week old female Wistars. The mobilized group had significantly greater number of cfus/ml compared to controls, p = 0.029. The isolated cells expressed 1.8% CD34, 35% CD45, 61% CD29, 78% CD90, and differentiated into osteoblasts but not into adipocytes. The fracture gap in animals treated with VEGF and AMD3100 showed increased bone volume; 5.22 ± 1.7 µm3 and trabecular thickness 0.05 ± 0.01 µm compared with control animals (4.3 ± 3.1 µm3 , 0.04 ± 0.01 µm, respectively). Radiographic scores quantifying fracture healing (RUST) showed that the animals in the mobilization group had a higher healing score compared to controls (9.6 vs. 7.7). Histologically, mobilization resulted in significantly lower group variability in bone formation (p = 0.032) and greater amounts of bone and less fibrous tissue than the control group. Clinical significance: This pre-clinical study demonstrates a beneficial effect of endogenous MSC mobilization on fracture healing, which may have translation potential to prevent or treat clinical fractures at risk of delayed or non-union fractures. © 2018 The Authors. Journal of Orthopaedic Research® Published by Wiley Periodicals, Inc. on behalf of Orthopaedic Research Society. J Orthop Res 37:1294-1302, 2019.


Assuntos
Fraturas do Fêmur/terapia , Consolidação da Fratura/efeitos dos fármacos , Compostos Heterocíclicos/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/farmacologia , Animais , Movimento Celular/efeitos dos fármacos , Quimiocina CXCL12/fisiologia , Feminino , Fraturas do Fêmur/diagnóstico por imagem , Fraturas do Fêmur/fisiopatologia , Células-Tronco Mesenquimais/fisiologia , Ratos , Ratos Wistar , Receptores CXCR4/antagonistas & inibidores , Receptores CXCR4/fisiologia , Microtomografia por Raio-X
6.
Biochim Biophys Acta Mol Cell Res ; 1866(2): 296-304, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30502369

RESUMO

Bone marrow (BM) is a source of mesenchymal stromal cells (MSCs) and endothelial progenitor cells (EPCs). MSCs provide a specific niche in the BM and biological features of EPCs may be changed with this niche. Stromal cell-derived factor 1 (SDF-1) secreted from primary BM-MSCs and biological features of this niche on EPC development are still yet to be understood. The aim of this study was to evaluate the role of SDF-1 produced by MSCs on EPC development. We applied the CRISPR/Cas9 system for the knock-out of the SDF-1 gene in BM-derived MSCs. BM-derived EPCs were then cocultured with MSCsSDF-1-/- or MSCsSDF-1+/+ to identify the role of MSC-derived SDF-1α on proliferation, migration and angiogenic activity of EPCs. Next, pre-expanded EPCs were harvested and co-transplanted with MSCsSDF-1-/- or MSCsSDF-1+/+ into sublethally irradiated mice to analyze the potency of these cells for marrow reconstitution. Our results revealed that proliferation, colony formation, migration and angiogenic activity of EPCs was significantly increased after coculture with MSCsSDF-1+/+. We also found that co-transplantation of EPCs with MSCsSDF-1+/+, in contrast to MSCsSDF-1-/-, into irradiated mice resulted in marrow repopulation and hematologic recovery, leading to improved survival of transplanted mice. In conclusions, MSC-derived SDF-1 niche plays an important role in the development of EPCs and this niche is essential for bone marrow repopulation by these cells and can enhance the efficiency of EPC therapy for ischemic diseases.


Assuntos
Plasticidade Celular/fisiologia , Quimiocina CXCL12/metabolismo , Quimiocina CXCL12/fisiologia , Animais , Células da Medula Óssea/metabolismo , Transplante de Medula Óssea/métodos , Movimento Celular , Proliferação de Células , Células Progenitoras Endoteliais/metabolismo , Masculino , Células-Tronco Mesenquimais/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Invasividade Neoplásica , Neovascularização Fisiológica
7.
BMB Rep ; 52(1): 42-46, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30526772

RESUMO

Cellular senescence, a process of cell proliferation arrest in response to various stressors, has been considered to be important factor in age-related disease. Identification of senescent cells in tissues is limited and the role of senescent cells is poorly understood. Recently however, several studies showed the characterization of senescent cells in various pathologic conditions and the role of senescent cells in disease progression is becoming important. Senescent cells are growth-arrested cells, however, the senescence associated secretory phenotype (SASP) of senescent cells could modify the tissues' microenvironment. Here, we discuss the progress and understanding of the role of senescent cells in tissues of pathologic conditions and discuss the development of new therapeutic paradigms, such as senescent cells-targeted therapy. [BMB Reports 2019; 52(1): 42-46].


Assuntos
Senescência Celular/fisiologia , Neoplasias/fisiopatologia , Animais , Carcinogênese/metabolismo , Proliferação de Células , Transformação Celular Neoplásica , Quimiocina CXCL12/fisiologia , Humanos , Fenótipo
8.
Oncogene ; 38(1): 73-87, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30082913

RESUMO

Primary glioblastoma is the most frequent human brain tumor in adults and is generally fatal due to tumor recurrence. We previously demonstrated that glioblastoma-initiating cells invade the subventricular zones and promote their radio-resistance in response to the local release of the CXCL12 chemokine. In this work, we show that the mitotic Aurora A kinase (AurA) is activated through the CXCL12-CXCR4 pathway in an ERK1/2-dependent manner. Moreover, the CXCL12-ERK1/2 signaling induces the expression of Ajuba, the main cofactor of AurA, which allows the auto-phosphorylation of AurA.We show that AurA contributes to glioblastoma cell survival, radio-resistance, self-renewal, and proliferation regardless of the exogenous stimulation with CXCL12. On the other hand, AurA triggers the CXCL12-mediated migration of glioblastoma cells in vitro as well as the invasion of the subventricular zone in xenograft experiments. Moreover, AurA regulates cytoskeletal proteins (i.e., Actin and Vimentin) and favors the pro-migratory activity of the Rho-GTPase CDC42 in response to CXCL12. Altogether, these results show that AurA, a well-known kinase of the mitotic machinery, may play alternative roles in human glioblastoma according to the CXCL12 concentration.


Assuntos
Aurora Quinase A/fisiologia , Neoplasias Encefálicas/enzimologia , Quimiocina CXCL12/fisiologia , Glioblastoma/enzimologia , Proteínas de Neoplasias/fisiologia , Animais , Neoplasias Encefálicas/patologia , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Sobrevivência Celular , Quimiocina CXCL12/farmacologia , Ativação Enzimática , Glioblastoma/patologia , Xenoenxertos , Humanos , Proteínas com Domínio LIM/biossíntese , Proteínas com Domínio LIM/genética , Ventrículos Laterais/patologia , Sistema de Sinalização das MAP Quinases , Camundongos , Invasividade Neoplásica , Fosforilação , Processamento de Proteína Pós-Traducional , Receptores CXCR4/fisiologia , Transdução de Sinais
9.
Invest Ophthalmol Vis Sci ; 59(12): 5201-5209, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-30372748

RESUMO

Purpose: Proper control of eye movements is critical to vision, but relatively little is known about the molecular mechanisms that regulate development and axon guidance in the ocular motor system or cause the abnormal innervation patterns (oculomotor synkinesis) seen in developmental disorders and after oculomotor nerve palsy. We developed an ex vivo slice assay that allows for live imaging and molecular manipulation of the growing oculomotor nerve, which we used to identify axon guidance cues that affect the oculomotor nerve. Methods: Ex vivo slices were generated from E10.5 IslMN-GFP embryos and grown for 24 to 72 hours. To assess for CXCR4 function, the specific inhibitor AMD3100 was added to the culture media. Cxcr4cko/cko:Isl-Cre:ISLMN-GFP and Cxcl12KO/KO:ISLMN-GFP embryos were cleared and imaged on a confocal microscope. Results: When AMD3100 was added to the slice cultures, oculomotor axons grew dorsally (away from the eye) rather than ventrally (toward the eye). Axons that had already exited the midbrain continued toward the eye. Loss of Cxcr4 or Cxcl12 in vivo caused misrouting of the oculomotor nerve dorsally and motor axons from the trigeminal motor nerve, which normally innervate the muscles of mastication, aberrantly innervated extraocular muscles in the orbit. This represents the first mouse model of trigeminal-oculomotor synkinesis. Conclusions: CXCR4/CXCL12 signaling is critical for the initial pathfinding decisions of oculomotor axons and their proper exit from the midbrain. Failure of the oculomotor nerve to innervate its extraocular muscle targets leads to aberrant innervation by other motor neurons, indicating that muscles lacking innervation may secrete cues that attract motor axons.


Assuntos
Quimiocina CXCL12/fisiologia , Doenças do Nervo Oculomotor/fisiopatologia , Nervo Oculomotor/anormalidades , Receptores CXCR4/fisiologia , Transdução de Sinais/fisiologia , Sincinesia/fisiopatologia , Núcleo Motor do Nervo Trigêmeo/fisiopatologia , Animais , Fármacos Anti-HIV/farmacologia , Axônios/patologia , Proteínas de Fluorescência Verde/metabolismo , Compostos Heterocíclicos/farmacologia , Imuno-Histoquímica , Hibridização In Situ , Camundongos , Camundongos Endogâmicos C57BL , Microscopia Confocal , Músculos Oculomotores/inervação , Nervo Oculomotor/efeitos dos fármacos , Técnicas de Cultura de Órgãos
10.
Adv Exp Med Biol ; 1099: 125-139, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30306520

RESUMO

Opioid analgesics remain the most effective and widely used analgesics for the management of moderate to severe pain, including cancer pain and chronic non-cancer pain. However, the efficacy of long-term opioid analgesics is attenuated by tolerance and/or hyperalgesia after long-term use, preventing adequate pain relief under stable opioid dosages for chronic pain patients. Classical neuron-centered concepts about tolerance, such as internalization of opioid receptors, upregulation of N-methyl-D-aspartate receptor function, or downregulation of glutamate transporter activity, can only partially explain the phenomenon of tolerance. Recent evidence revealing glial activation and upregulation of inflammatory mediators in the rodent central nervous system has confirmed the pivotal role of neuroinflammation in neuropathic pain or opioid tolerance, or both. However, human evidence is still sparse.Based on our clinical practice, we conducted translational research by investigating the cerebrospinal fluid (CSF) cytokine and chemokine profiles of opioid-tolerant patients after research ethic committee approval. CSF samples from opioid-tolerant patients and opioid-naive subjects were compared. We found CXCL1, CXCL12, and leukemia inhibitory factor (LIF) were significantly upregulated among the opioid-tolerant patients and positively correlated with the opioid dosage.We translated these findings back to lab animal experiment; after induction of tolerance by morphine infusion, the spinal cord expression of CXCL1, CXCL12, and LIF were all upregulated. Although CXCL1 and CXCL12 infusion alone did not affect baseline tail-flick latency, morphine analgesic efficacy dropped significantly after intrathecal infusion of CXCL1 and CXCL12. After establishing tolerance by intrathecal continuous infusion of morphine, tolerance development was accelerated by co-administration of CXCL1 and CXCL12. In parallel, the effect was attenuated by co-administration of CXCL1- or CXCL12-neutralizing antibody or concordant receptor antagonists.On the contrary, although chronic morphine administration still induced LIF upregulation in rat spinal cords, intrathecal injection of LIF potentiated the analgesic action of morphine and delayed the development of morphine tolerance. Upregulation of endogenously released LIF by long-term use of opioids might counterbalance the tolerance induction effects of other pro-inflammatory cytokines.CXCL1, CXCL12, and LIF are upregulated in both opioid-tolerant patients and rodents. The onset and extent of opioid tolerance were affected by modulating the intrathecal CXCL1/CXCR2, CXCL12/CXCR4, and LIF signaling and could be novel drug targets for the treatment of opioid tolerance.


Assuntos
Analgésicos Opioides/farmacologia , Quimiocina CXCL12/fisiologia , Quimiocina CXCL1/fisiologia , Tolerância a Medicamentos , Inflamação/fisiopatologia , Fator Inibidor de Leucemia/fisiologia , Animais , Humanos , Ratos , Medula Espinal/efeitos dos fármacos
11.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 36(4): 398-403, 2018 Aug 01.
Artigo em Chinês | MEDLINE | ID: mdl-30182568

RESUMO

OBJECTIVE: To observe the influence of LM609/AMD3100/CCX754 on chemotactic capability, cytoskeleton, and expression of integrin ανß3 protein of squamous cell carcinoma of head and neck (SCCHN) cell line PCI-13 induced by stromal cell-derived factor-1 (SDF-1) in vitro. METHODS: Migration assays, flow cytometry and immunofluorescence were used to observe the effects of SDF-1, LM609, AMD3100 and CCX754 on the migration, cytoskeleton and the expression of integrin ανß3 protein in PCI-13 cell lines. RESULTS: SDF-1 favored PCI-13 cell migration, pseudopod formation, and activities of integrin ανß3 phosphorylation. LM609, AMD3100, and CCX754 blocked all these effects. CONCLUSIONS: SDF-1 can induce metastatic SCCHN by integrin ανß3-CXC chemokine receptor (CXCR) 4/CXCR7 axi. LM609, AMD3100, and CCX754 and can reduce the regulation of SDF-1 on SCCHN activity.


Assuntos
Carcinoma de Células Escamosas , Quimiocina CXCL12 , Neoplasias de Cabeça e Pescoço , Metástase Neoplásica , Carcinoma de Células Escamosas de Cabeça e Pescoço , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patologia , Movimento Celular , Quimiocina CXCL12/fisiologia , Neoplasias de Cabeça e Pescoço/metabolismo , Neoplasias de Cabeça e Pescoço/patologia , Humanos , Integrinas , Receptores CXCR/metabolismo , Receptores CXCR4 , Carcinoma de Células Escamosas de Cabeça e Pescoço/metabolismo , Carcinoma de Células Escamosas de Cabeça e Pescoço/patologia , Células Estromais
12.
BMC Cancer ; 18(1): 741, 2018 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-30012106

RESUMO

BACKGROUND: The receptor CXCR4 and its ligand CXCL12 play crucial roles in breast cancer. Despite the fact that the spontaneous feline mammary carcinoma (FMC) is considered a suitable model for breast cancer studies, the importance of the CXCR4/CXCL12 axis in FMC is completely unknown. Therefore, this work aims to elucidate the role of CXCR4 and its ligand in the progression of FMC and metastatic disease. METHODS: CXCR4 and CXCL12 expression was analyzed by immunohistochemistry and immunofluorescence on primary tumors (PT), regional and distant metastases of female cats with mammary carcinoma and correlated with serum CXCL12 levels, tumor molecular subtypes and clinicopathological features. RESULTS: CXCR4 was more expressed in PT than in metastases (p = 0.0067), whereas CXCL12 was highly expressed in metastatic lesions located in liver and lung (p < 0.0001), as reported for human breast cancer. Moreover, cats with CXCR4 positive PT exhibited significantly lower serum CXCL12 levels than cats with CXCR4 negative mammary carcinomas (p = 0.0324). At metastatic lesions, HER2-overexpressing tumors presented higher CXCR4 expression than the other molecular tumor subtypes (p = 0.012) and significant differences in overall (p = 0.0147) and disease-free survival (p = 0.0279) curves between the cats with CXCL12 positive and CXCL12 negative tumors were found. Indeed, CXCL12 negative PT were associated with unfavorable prognosis in cats with HER2-overexpressing tumors. CONCLUSIONS: This work exposes part of the complex interaction between CXCR4 and CXCL12 in PT, but also in metastases of a breast cancer model. These findings could uncover novel therapeutic tools to be used in cats and humans.


Assuntos
Doenças do Gato/etiologia , Quimiocina CXCL12/fisiologia , Neoplasias Mamárias Animais/etiologia , Receptor ErbB-2/análise , Receptores CXCR4/fisiologia , Animais , Doenças do Gato/imunologia , Doenças do Gato/patologia , Gatos , Quimiocina CXCL12/análise , Feminino , Neoplasias Mamárias Animais/imunologia , Neoplasias Mamárias Animais/patologia , Metástase Neoplásica , Prognóstico , Receptores CXCR4/análise
13.
Mol Biol Rep ; 45(5): 741-750, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29926386

RESUMO

CXCL12/CXCR4 signaling has been implicated in breast carcinogenesis, and genetic polymorphisms in these molecules have been associated with different types of cancer. The present study analyzed genetic polymorphisms in CXCL12 (rs1801157, G > A) and CXCR4 (rs2228014, C > T) and CXCR4 immunostaining in tumor tissues from patients with triple negative breast cancer (TNBC) aiming to evaluate their possible role in its' susceptibility and prognosis. Genetic polymorphisms were analyzed in 59 TNBC patients and 150 control women; age-adjusted logistic regression showed no association when variants were considered in isolation; however, a statistically significant interaction was noted for heterozygosis for both allelic variants increasing the odds for TNBC (CXCL12-GA by CXCR4-CT: OR 7.23; 95% CI 1.15-45.41; p = 0.035). CXCL12 polymorphism was correlated negatively with proliferation index (Ki67) (Tau-b = - 0.406; p = 0.006). CXCR4 immunostaining was evaluated in 37 TNBC patients (22 with paired tumor-normal adjacent tissue). CXCR4 was detected more intensely in cell cytoplasm than in membrane, and was more expressed in tumor than in normal adjacent tissues, although not statistically significant. CXCR4 expression on the membrane of tumor cells was correlated positively with histopathological grade (Tau-b = 0.271; p = 0.036) and negatively with lymph node metastasis (Tau-b = - 0.478; p = 0.036). The present study indicates that CXCL12 and CXCR4 polymorphisms and CXCR4 immunostaining might have susceptibility and prognostic roles in TNBC pathogenesis.


Assuntos
Quimiocina CXCL12/genética , Quimiocina CXCL12/fisiologia , Receptores CXCR4/genética , Neoplasias de Mama Triplo Negativas/genética , Adulto , Idoso , Alelos , Biomarcadores Tumorais/genética , Feminino , Frequência do Gene/genética , Estudos de Associação Genética , Predisposição Genética para Doença/genética , Humanos , Metástase Linfática , Pessoa de Meia-Idade , Polimorfismo Genético , Polimorfismo de Nucleotídeo Único/genética , Prognóstico , Receptores CXCR4/fisiologia , Neoplasias de Mama Triplo Negativas/fisiopatologia
14.
Gastroenterology ; 155(3): 880-891.e8, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29909021

RESUMO

BACKGROUND & AIMS: Immunotherapies are ineffective against pancreatic cancer. We investigated whether the activity of nuclear factor (NF)κB in pancreatic stromal cells contributes to an environment that suppresses antitumor immune response. METHODS: Pancreata of C57BL/6 or Rag1-/- mice were given pancreatic injections of a combination of KrasG12D/+; Trp53 R172H/+; Pdx-1cre (KPC) pancreatic cancer cells and pancreatic stellate cells (PSCs) extracted from C57BL/6 (control) or mice with disruption of the gene encoding the NFκB p50 subunit (Nfkb1 or p50-/- mice). Tumor growth was measured as an endpoint. Other mice were given injections of Lewis lung carcinoma (LLC) lung cancer cells or B16-F10 melanoma cells with control or p50-/- fibroblasts. Cytotoxic T cells were depleted from C57BL/6 mice by administration of antibodies against CD8 (anti-CD8), and growth of tumors from KPC cells, with or without control or p50-/- PSCs, was measured. Some mice were given an inhibitor of CXCL12 (AMD3100) and tumor growth was measured. T-cell migration toward cancer cells was measured using the Boyden chamber assay. RESULTS: C57BL/6 mice coinjected with KPC cells (or LLC or B16-F10 cells) and p50-/- PSCs developed smaller tumors than mice given injections of the cancer cells along with control PSCs. Tumors that formed when KPC cells were injected along with p50-/- PSCs had increased infiltration by activated cytotoxic T cells along with decreased levels of CXCL12, compared with tumors grown from KPC cells injected along with control PSCs. KPC cells, when coinjected with control or p50-/- PSCs, developed the same-size tumors when CD8+ T cells were depleted from C57BL/6 mice or in Rag1-/- mice. The CXCL12 inhibitor slowed tumor growth and increased tumor infiltration by cytotoxic T cells. In vitro expression of p50 by PSCs reduced T-cell migration toward and killing of cancer cells. When cultured with cancer cells, control PSCs expressed 10-fold higher levels of CXCL12 than p50-/- PSCs. The CXCL12 inhibitor increased migration of T cells toward KPC cells in culture. CONCLUSIONS: In studies of mice and cell lines, we found that NFκB activity in PSCs promotes tumor growth by increasing expression of CXCL12, which prevents cytotoxic T cells from infiltrating the tumor and killing cancer cells. Strategies to block CXCL12 in pancreatic tumor cells might increase antitumor immunity.


Assuntos
Quimiocina CXCL12/fisiologia , Linfócitos do Interstício Tumoral/fisiologia , NF-kappa B/fisiologia , Neoplasias Pancreáticas/metabolismo , Células Estreladas do Pâncreas/metabolismo , Linfócitos T Citotóxicos/fisiologia , Animais , Carcinogênese/metabolismo , Linhagem Celular Tumoral , Imunidade Celular , Camundongos , Camundongos Endogâmicos C57BL , Neoplasias Pancreáticas/imunologia , Células Estreladas do Pâncreas/imunologia , Regulação para Cima
15.
PLoS One ; 13(6): e0198789, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29920526

RESUMO

The CXCR4/CXCL12 axis has been extensively associated with different types of cancer correlating with higher aggressiveness and metastasis. In diffuse large B-cell lymphoma (DLBCL), the expression of the chemokine receptor CXCR4 is involved in the dissemination of malignant B cells and is a marker of poor prognosis. CXCR7 is a chemokine receptor that binds to the same ligand as CXCR4 and regulates de CXCR4-CXCL12 axis. These findings together with the report of CXCR7 prognostic value in several tumor types, led us to evaluate the expression of CXCR7 in diffuse large B-cell lymphoma biopsies. Here, we describe that CXCR7 receptor is an independent prognostic factor that associates with good clinical outcome. Moreover, the expression of CXCR7 associates with increased survival in CXCR4+ but not in CXCR4- DLBCL patients. Thus, the combined immunohistochemical evaluation of both CXCR7 and CXCR4 expression in DLBCL biopsies may improve their prognostic value as single markers. Finally, we show that CXCR7 overexpression in vitro is able to diminish DLBCL cell survival and increase their sensitivity to antitumor drugs. Hence, further studies on the CXCR7 receptor may establish its role in DLBCL and the molecular mechanisms that modulate CXCR4 activity.


Assuntos
Regulação Neoplásica da Expressão Gênica , Linfoma Difuso de Grandes Células B/genética , Proteínas de Neoplasias/biossíntese , Receptores CXCR4/análise , Receptores CXCR/biossíntese , Adulto , Idoso , Biomarcadores Tumorais , Biópsia , Linhagem Celular Tumoral , Quimiocina CXCL12/fisiologia , Resistencia a Medicamentos Antineoplásicos/genética , Feminino , Humanos , Estimativa de Kaplan-Meier , Linfoma Difuso de Grandes Células B/metabolismo , Linfoma Difuso de Grandes Células B/mortalidade , Linfoma Difuso de Grandes Células B/patologia , Masculino , Pessoa de Meia-Idade , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/fisiologia , Prognóstico , Modelos de Riscos Proporcionais , Receptores CXCR/genética , Receptores CXCR/fisiologia
16.
Eur J Clin Invest ; 48 Suppl 2: e12949, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29734477

RESUMO

BACKGROUND: Chemokines play a critical role in orchestrating the distribution and trafficking of neutrophils in homeostasis and disease. RESULTS: The CXCR4/CXCL12 chemokine axis has been identified as a central regulator of these processes. CONCLUSION: In this review, we focus on the role of CXCR4/CXCL12 chemokine axis in regulating neutrophil release from the bone marrow and the trafficking of senescent neutrophils back to the bone marrow for clearance under homeostasis and disease. We also discuss the role of CXCR4 in fine-tuning neutrophil responses in the context of inflammation.


Assuntos
Homeostase/fisiologia , Neutrófilos/fisiologia , Receptores CXCR4/fisiologia , Animais , Benzilaminas/farmacologia , Medula Óssea/fisiologia , Sobrevivência Celular/fisiologia , Quimiocina CXCL12/genética , Quimiocina CXCL12/fisiologia , Proteína HMGB1/fisiologia , Fármacos Hematológicos/farmacologia , Compostos Heterocíclicos/farmacologia , Humanos , Imidazóis/farmacologia , Síndromes de Imunodeficiência/genética , Inflamação/fisiopatologia , Camundongos , Mutação/fisiologia , Neutrófilos/efeitos dos fármacos , Fragmentos de Peptídeos/farmacologia , Receptores CXCR4/antagonistas & inibidores , Albumina Sérica/farmacologia , Baço/fisiologia , Verrugas/genética
17.
Acta Oncol ; 57(9): 1134-1142, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29771176

RESUMO

Medulloblastoma (MB) is the most common malignant brain tumor occurring in children, and although high long-term survival rates have been reached with current therapeutic protocols, several neurological injuries are still observed among survivors. It has been shown that the development of MB is highly dependent on the microenvironment surrounding it and that the CXCL12 chemokine and its receptor, CXCR4 and the Sonic Hedgehog (SHH) pathway are crucial for cerebellar development, coordinating proliferation and migration of embryonic cells and malfunctions in these axes can lead to MB development. Indeed, the concomitant overactivation of these axes was suggested to define a new MB molecular subgroup. New molecules are being studied, aiming to inhibit either CXCR4 or the SHH pathways and have been tested in preclinical settings for the treatment of cancers. The use of these molecules could improve MB treatment and save patients from aggressive surgery, chemotherapy and radiotherapy regimens, which are responsible for severe neurological consequences. This review aims to summarize current data about the experimental inhibition of CXCR4 and SHH pathways in MB and its potential implications in treatment of this cancer.


Assuntos
Antineoplásicos/uso terapêutico , Neoplasias Cerebelares/terapia , Quimiocina CXCL12/fisiologia , Proteínas Hedgehog/fisiologia , Meduloblastoma/terapia , Terapia de Alvo Molecular/métodos , Receptores CXCR4/fisiologia , Neoplasias Cerebelares/patologia , Quimiocina CXCL12/antagonistas & inibidores , Proteínas Hedgehog/antagonistas & inibidores , Humanos , Meduloblastoma/patologia , Receptores CXCR4/antagonistas & inibidores , Transdução de Sinais/fisiologia
18.
Eur J Med Chem ; 149: 30-44, 2018 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-29494843

RESUMO

The important roles of the CXCL12/CXCR4 axis in numerous pathogenic pathways involving HIV infection and cancer metastasis make the CXCR4 receptor an attractive target for the development of therapeutic agents. Through scaffold hybridization of a few known CXCR4 antagonists, a series of novel aminopyrimidine derivatives was developed. Compound 3 from this new scaffold demonstrates excellent binding affinity with CXCR4 receptor (IC50 = 54 nM) and inhibits CXCL12 induced cytosolic calcium increase (IC50 = 2.3 nM). Furthermore, compound 3 possesses good physicochemical properties (MW 353, clogP 2.0, PSA 48, pKa 6.7) and exhibits minimal hERG and CYP isozyme (e.g. 3A4, 2D6) inhibition. Collectively, these results strongly support further optimization of this novel scaffold to develop better CXCR4 antagonists.


Assuntos
Desenho de Drogas , Pirimidinas/química , Receptores CXCR4/antagonistas & inibidores , Cálcio/metabolismo , Quimiocina CXCL12/fisiologia , Humanos , Ligação Proteica , Pirimidinas/síntese química , Pirimidinas/farmacologia , Relação Estrutura-Atividade
19.
J Theor Biol ; 444: 93-99, 2018 05 07.
Artigo em Inglês | MEDLINE | ID: mdl-29470991

RESUMO

Collective cell migration is an integral part of organismal development. We consider migration of the zebrafish primordium during development of the posterior lateral line, a sensory system that detects water movement patterns. Experiments have shown that the chemokine ligand CXCL12a and its receptors CXCR4b and CXCR7b are key players for driving migration of the primordium, while FGF signaling helps maintain cohesion. In this work, we formulate a mathematical model of a laser ablated primordium separated into two smaller cell collectives: a leading collective that responds to local CXCL12a levels and a trailing collective that migrates up a local FGF gradient. Our model replicates recent experimental results, while also predicting a "runaway" behavior when FGF gradient response is inhibited. We also use our model to estimate diffusion coefficients of CXCL12a and FGF in the lateral line.


Assuntos
Movimento Celular/fisiologia , Sistema da Linha Lateral/citologia , Modelos Teóricos , Peixe-Zebra/anatomia & histologia , Animais , Comunicação Celular/fisiologia , Quimiocina CXCL12/metabolismo , Quimiocina CXCL12/fisiologia , Difusão , Desenvolvimento Embrionário , Fatores de Crescimento de Fibroblastos/metabolismo , Fatores de Crescimento de Fibroblastos/fisiologia , Sistema da Linha Lateral/embriologia , Sistema da Linha Lateral/crescimento & desenvolvimento , Peixe-Zebra/embriologia , Proteínas de Peixe-Zebra/metabolismo , Proteínas de Peixe-Zebra/fisiologia
20.
Cancer Res ; 78(8): 2026-2039, 2018 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-29431639

RESUMO

There is evidence that cancer stem-like cells (CSC) and neuroendocrine behavior play critical roles in the pathogenesis and clinical course of metastatic castration-resistant prostate cancer (m-CRPC). However, there is limited mechanistic understanding of how CSC and neuroendocrine phenotypes impact the development of m-CRPC. In this study, we explored the role of the intracellular chemokine CXCL12γ in CSC induction and neuroendocrine differentiation and its impact on m-CRPC. CXCL12γ expression was detected in small-cell carcinoma of metastatic tissues and circulating tumor cells from m-CRPC patients and in prostate cancer cells displaying an neuroendocrine phenotype. Mechanistic investigations demonstrated that overexpression of CXCL12γ induced CSC and neuroendocrine phenotypes in prostate cancer cells through CXCR4-mediated PKCα/NFκB signaling, which promoted prostate tumor outgrowth, metastasis, and chemoresistance in vivo Together, our results establish a significant function for CXCL12γ in m-CRPC development and suggest it as a candidate therapeutic target to control aggressive disease.Significance: Expression of CXCL12γ induces the expression of a cancer stem cell and neuroendocrine phenotypes, resulting in the development of aggressive m-CRPC. Cancer Res; 78(8); 2026-39. ©2018 AACR.


Assuntos
Neoplasias Ósseas/secundário , Reprogramação Celular , Quimiocina CXCL12/fisiologia , Células-Tronco Neoplásicas/patologia , Células Neuroendócrinas/patologia , Neoplasias de Próstata Resistentes à Castração/patologia , Animais , Linhagem Celular Tumoral , Proliferação de Células/fisiologia , Feminino , Xenoenxertos , Humanos , Masculino , Camundongos SCID , NF-kappa B/metabolismo , Fenótipo , Proteína Quinase C-alfa/metabolismo , Receptores CXCR4/metabolismo , Transdução de Sinais
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