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BACKGROUND: Cyclin-dependent kinase 9 (CDK9) expression and autophagy in colorectal cancer (CRC) tissues has not been widely studied. CDK9, a key regulator of transcription, may influence the occurrence and progression of CRC. The expression of autophagy-related genes BECN1 and drug resistance factor ABCG2 may also play a role in CRC. Under normal physiological conditions, autophagy can inhibit tumorigenesis, but once a tumor forms, autophagy may promote tumor growth. Therefore, understanding the relationship between autophagy and cancer, particularly how autophagy promotes tumor growth after its formation, is a key motivation for this research. AIM: To investigate the relationship between CDK9 expression and autophagy in CRC, assess differences in autophagy between left and right colon cancer, and analyze the associations of autophagy-related genes with clinical features and prognosis. METHODS: We collected tumor tissues and paracarcinoma tissues from colon cancer patients with liver metastasis to observe the level of autophagy in tissues with high levels of CDK9 and low levels of CDK9. We also collected primary tissue from left and right colon cancer patients with liver metastasis to compare the autophagy levels and the expression of BECN1 and ABCG2 in the tumor and paracarcinoma tissues. RESULTS: The incidence of autophagy and the expression of BECN1 and ABCG2 were different in left and right colon cancer, and autophagy might be involved in the occurrence of chemotherapy resistance. Further analysis of the relationship between the expression of autophagy-related genes CDK9, ABCG2, and BECN1 and the clinical features and prognosis of colorectal cancer showed that the high expression of CDK9 indicated a poor prognosis in colorectal cancer. CONCLUSION: This study laid the foundation for further research on the combination of CDK9 inhibitors and autophagy inhibitors in the treatment of patients with CRC.
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BACKGROUND: Esophageal squamous cell carcinoma (ESCC) is a deadly malignancy with limited treatment options. Deubiquitinases (DUBs) have been confirmed to play a crucial role in the development of malignant tumors. JOSD2 is a DUB involved in controlling protein deubiquitination and influencing critical cellular processes in cancer. AIM: To investigate the impact of JOSD2 on the progression of ESCC. METHODS: Bioinformatic analyses were employed to explore the expression, prognosis, and enriched pathways associated with JOSD2 in ESCC. Lentiviral transduction was utilized to manipulate JOSD2 expression in ESCC cell lines (KYSE30 and KYSE150). Functional assays, including cell proliferation, colony formation, drug sensitivity, migration, and invasion, were performed, revealing the impact of JOSD2 on ESCC cell lines. JOSD2's role in xenograft tumor growth and drug sensitivity in vivo was also assessed. The proteins that interacted with JOSD2 were identified using mass spectrometry. RESULTS: Preliminary research indicated that JOSD2 was highly expressed in ESCC tissues, which was associated with poor prognosis. Further analysis demonstrated that JOSD2 was upregulated in ESCC cell lines compared to normal esophageal cells. JOSD2 knockdown inhibited ESCC cell activity, including proliferation and colony-forming ability. Moreover, JOSD2 knockdown decreased the drug resistance and migration of ESCC cells, while JOSD2 overexpression enhanced these phenotypes. In vivo xenograft assays further confirmed that JOSD2 promoted tumor proliferation and drug resistance in ESCC. Mechanistically, JOSD2 appears to activate the MAPK/ERK and PI3K/AKT signaling pathways. Mass spectrometry was used to identify crucial substrate proteins that interact with JOSD2, which identified the four primary proteins that bind to JOSD2, namely USP47, IGKV2D-29, HSP90AB1, and PRMT5. CONCLUSION: JOSD2 plays a crucial role in enhancing the proliferation, migration, and drug resistance of ESCC, suggesting that JOSD2 is a potential therapeutic target in ESCC.
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Neoplasias Esofágicas , Carcinoma de Células Escamosas de Esófago , Humanos , Carcinoma de Células Escamosas de Esófago/patología , Neoplasias Esofágicas/patología , Fosfatidilinositol 3-Quinasas/metabolismo , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular , Enzimas Desubicuitinizantes/genética , Regulación Neoplásica de la Expresión Génica , Proteína-Arginina N-MetiltransferasasRESUMEN
Agmatinase (AGMAT) is an enzyme that hydrolyzes agmatine to putrescine and urea. In this study, we explored the functions of AGMAT in colorectal cancer (CRC). By performing gain-of-function and loss-of-function experiments, we investigated the roles of AGMAT in proliferation, cell cycle progression, and apoptosis of CRC cells. We also established a colitis-associated colorectal cancer model by challenging mice with azoxymethane (AOM) and dextran sodium sulfate (DSS), and we subsequently silenced AGMAT expression in mice by adeno-associated virus 9 (AAV9)-mediated delivery of short hairpin RNA (shRNA). In vitro experiments showed that overexpression of AGMAT accelerated the proliferation and inhibited the apoptosis of CRC cells, and AGMAT knockdown exhibited the opposite effects. Interestingly, the oncogenic transcription factor, c-Myc, could bind to the AGMAT promoter and transcriptionally increase AGMAT expression in CRC cells. Additionally, c-Myc and AGMAT were upregulated in the colon of AOM/DSS-treated mice, and AGMAT silencing significantly mitigated colitis in AOM/DSS-treated mice, as evidenced by the increased colon length, attenuated crypt damage, and reduced levels of inflammatory indicators (myeloperoxidase, interleukin-6, tumor necrosis factor-α, inducible nitric oxide synthase, and phosphorylated p65) in colon tissues. Notably, AGMAT silencing decreased both the number and size of tumors, reduced expression of proliferating cell nuclear antigen, and inhibited phosphorylation of protein kinase B (AKT) and extracellular signal-regulated kinase in the colon of AOM/DSS-treated mice. Overall, we determined that AGMAT facilitates tumor progression in CRC. Our findings will be helpful in the search for potential therapeutic targets for CRC.
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Neoplasias Asociadas a Colitis , Neoplasias Colorrectales , Ratones , Animales , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/terapia , Dependovirus/genética , Inflamación , Transformación Celular Neoplásica , Carcinogénesis/genética , Modelos Animales de Enfermedad , Ratones Endogámicos C57BLRESUMEN
This study was designed and conducted to clarify the impact of RNF128 expression on malignant biological behaviors of colorectal cancer (CRC) cells and the underlying mechanism. The expression of RNF128 in CRC tissues was analyzed using mRNA sequencing data of TCGA database and was validated by Western blot assay. The experimental studies on biological functions of RNF128 in vitro were conducted to assess its impact on the proliferation, apoptosis, and metastasis of CRC cells. Furthermore, tumor xenograft models in nude mice were established to investigate the relationship between RNF128 expression and tumor growth in vivo. The expression levels of both RNF128 mRNA and protein were significantly increased in CRC tissues (p < .001). The knockdown of RNF128 markedly suppressed the malignant phenotype of HCT116 and SW480 cells in vitro, including cell growth, antiapoptosis, migration, and invasion (p < .001). On the other hand, knockdown of RNF128 exerted a remarkable effect on the growth inhibition of tumor xenografts in vivo (p < .001). Further investigation revealed that RNF128 knockdown lead to a significant decrease in the expression of p-AKT and p-PI3K protein. More importantly, the proliferative, antiapoptotic, metastatic abilities of RNF128-knockdown cells were markedly increased by 740 Y-P treatment (p < .001). These findings further suggested that PI3K/AKT signaling pathway played a key role in RNF128-mediated aggressive phenotype of CRC cells. RNF128 functions as a tumor promoter in the pathogenesis of CRC via regulating PI3K/AKT pathway, and it could be a valuable target for CRC treatment.
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Neoplasias Colorrectales , Fosfatidilinositol 3-Quinasas , Animales , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Neoplasias Colorrectales/metabolismo , Regulación Neoplásica de la Expresión Génica , Humanos , Ratones , Ratones Desnudos , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , ARN Mensajero , Transducción de Señal/genética , Ubiquitina-Proteína Ligasas/genética , Ubiquitina-Proteína Ligasas/metabolismoRESUMEN
Postsynaptic NMDARs at spinal synapses are required for postsynaptic long-term potentiation and chronic pain. However, how presynaptic NMDARs (PreNMDARs) in spinal nociceptor terminals control presynaptic plasticity and pain hypersensitivity has remained unclear. Here we report that PreNMDARs in spinal nociceptor terminals modulate synaptic transmission in a nociceptive tone-dependent manner. PreNMDARs depresses presynaptic transmission in basal state, while paradoxically causing presynaptic potentiation upon injury. This state-dependent modulation is dependent on Ca2+ influx via PreNMDARs. Small conductance Ca2+-activated K+ (SK) channels are responsible for PreNMDARs-mediated synaptic depression. Rather, tissue inflammation induces PreNMDARs-PKG-I-dependent BDNF secretion from spinal nociceptor terminals, leading to SK channels downregulation, which in turn converts presynaptic depression to potentiation. Our findings shed light on the state-dependent characteristics of PreNMDARs in spinal nociceptor terminals on modulating nociceptive transmission and revealed a mechanism underlying state-dependent transition. Moreover, we identify PreNMDARs in spinal nociceptor terminals as key constituents of activity-dependent pain sensitization.
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Dolor Crónico/fisiopatología , Nociceptores/metabolismo , Terminales Presinápticos/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Animales , Factor Neurotrófico Derivado del Encéfalo/genética , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Calcio/metabolismo , Dolor Crónico/genética , Dolor Crónico/metabolismo , Proteína Quinasa Dependiente de GMP Cíclico Tipo I/genética , Proteína Quinasa Dependiente de GMP Cíclico Tipo I/metabolismo , Ganglios Espinales/citología , Ganglios Espinales/fisiología , Inflamación , Potenciación a Largo Plazo , Depresión Sináptica a Largo Plazo , Ratones , Ratones Transgénicos , Sustancia Gris Periacueductal/citología , Sustancia Gris Periacueductal/fisiología , Canales de Potasio Calcio-Activados/genética , Canales de Potasio Calcio-Activados/metabolismo , Receptores de N-Metil-D-Aspartato/genética , Transmisión SinápticaRESUMEN
As an important part of the soil microbial system, fungi can clearly indicate changes in the soil environment.Human activities in the city can easily affect the soil condition, so the phenomenon of artificial heavy metal enrichment often appears in urban soil. The objective of this study was to analyze the fungal community structure in different urban functional areas and to determine the effect of heavy metal content in different urban functional areas on the soil fungal community structure. This study provides theoretical basis for protecting and repairing the urban soil ecosystem and transforming and improving urban environmental quality. Soil samples from eight sampling sites in five functional areas of Beibei District in Chongqing were taken as the research objects. The diversity and community structure of fungi in soil were studied using high-throughput sequencing technology. The content of Cd and Hg in the soil of different functional areas in Beibei District exceeded the environmental background value of Chongqing. The 20-40 cm and 40-60 cm soil layers of JD were slightly polluted. The 20-40 cm soil layer and 0-20 cm soil layer of JLD and ZYY, respectively, were in the alerting state of heavy metal pollution. The Sobs index, Chao 1 index, and Ace index of most sampling points decreased with the increase in soil depth. The NMDS analysis showed that the composition of fungal communities between the 0-20 cm and 20-40 cm soil depths in both JD and ZYY were quite different. From the perspective of community composition, Ascomycota was the most abundant phylum in the soil, followed by unclassified _k _Fungi and Basidiomycota. At the genus level, unclassified_k_Fungi, unclassified_p_Ascomycota, unclassified_o_Sordariales, Scopuloides, Robillarda, and Dactylonectria had higher abundances. The redundancy analysis (RDA) showed that Pb and Zn had the greatest effect on the samples, and the effect on the fungal community structure was significantly different. This study has deepened the understanding of the relationship between the content of heavy metals in different urban functional areas and the structure of fungal communities and has provided a scientific basis for the rational use and planning of urban soil.
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Metales Pesados , Micobioma , Contaminantes del Suelo , China , Ecosistema , Humanos , Metales Pesados/análisis , Suelo , Microbiología del Suelo , Contaminantes del Suelo/análisisRESUMEN
BACKGROUND: Colorectal cancer (CRC) is one of the most common cancers. The aim of our study was to explore its related mutations, identify novel mutation markers, and construct predictive models for postoperative CRC patients, so as to provide evidence for the diagnosis, treatment, and prognosis of CRC. METHODS: A total 50 CRC patients were collected, and the mutations in tissue samples were detected through next-generation sequencing (NGS). Meanwhile, 246 CRC cases with complete mutation data were downloaded from The Cancer Genome Atlas (TCGA) database. Afterwards, the co-mutations in both clinical and TCGA cohorts were identified, and the high-frequency mutation genes were selected. Subsequently, functional enrichment analysis was performed, and overall survival (OS) and progression-free survival (PFS) predictive models were constructed. RESULTS: In all, 18 out of 238 co-mutation genes mutated in at least 20% of the samples and were selected out as common high-frequency mutation genes. They were significantly enriched in 460 Gene Ontology (GO) terms and 87 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways (P<0.05), which were closely related to the occurrence and development of CRC. Among the 18 genes, NOTCH3, histone lysine methyltransferase 2C (KMT2C), and cAMP-response element binding protein-BP (CREBBP) were respectively associated with tumor position, stage, and PFS (P<0.05), and could be considered as potential biomarkers of CRC. Finally, OS and PFS predictive models were constructed and verified using the 50 clinical cases, with both models demonstrating high fitting degrees useful for predicting the OS and PFS of CRC patients. CONCLUSIONS: NOTCH3, KMT2C, and CREBBP were found to be prospective biomarkers for the diagnosis and prognosis of CRC. The prognosis prediction models had high sensitivity and could be used to predict the OS and PFS of CRC patients.
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OBJECTIVE@#The incidence of colorectal stromal tumor is low among digestive tract tumors, therefore the literatures about clinicopathological features and prognosis of colorectal stromal tumor are few at home and abroad. In this study, we performed survival analyses for colorectal stromal tumor. The nomogram made by prognostic factors provided basis for evaluation of prognosis.@*METHODS@#The clinico-pathological and prognostic data of colorectal stromal tumor between January 1992 and December 2015 were collected from the surveillance, epidemiology, and end results (SEER) database. The survival analyses were made by SPSS 24.0 software. The nomogram and calibration curve were made by RMS package in R 3.5.2 software.@*RESULTS@#In the study, 546 patients with colorectal stromal tumor were included. The median age of onset was 64 years. The regional lymph node metastasis (LNM) rate was 9.4%. The multivariate Cox regression analyses of the 546 cases showed that the older age of onset (>64 years), single or divorce, colon tumor (compared with rectal tumor), non-surgery, high histological grade, LNM and distant metastasis were associated with worse cancer specific survival (CSS) and overall survival (OS), P < 0.05 for all. The treatment district was independent prognostic factor of OS (P = 0.027). The C-index of independent prognostic factors predicting CSS and OS probability were 0.76 (95%CI: 0.72-0.80) and 0.75 (95%CI: 0.72-0.78), respectively. Multivariate analyses were further carried out in the 174 patients with definite histological grade and tumor location, which revealed that the age of onset, histological grade, surgery or not were independent prognostic factors of CSS and OS (P < 0.05 for all). Tumor location was associated with CSS (P = 0.041) but not OS (P = 0.057) among the 174 cases. Four independent prognostic factors influencing the 174 patients' prognosis were used to make nomogram for predicting survival probability of 546 cases. The C-index of four prognostic factors predicting probability of CSS and OS of the 546 cases were separately 0.71 (95%CI: 0.66-0.75) and 0.73 (95%CI: 0.70-0.77). The nomogram had more accuracy for predicting OS probability of colorectal stromal tumors.@*CONCLUSION@#The prognosis of colorectal stromal tumor was affected by multiple clinicopathological factors. The nomogram provided the basis for predicting the survival probability of patients with colorectal stromal tumor.
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Anciano , Humanos , Persona de Mediana Edad , Neoplasias Colorrectales , Estadificación de Neoplasias , Pronóstico , Programa de VERFRESUMEN
BACKGROUND: Calponin is an actin filament-associated regulatory protein originally identified in smooth muscle cells. Three homologous have been identified in vertebrate species, but little functional characterization of potential activities in cancer has been performed. In this study, we determined the levels of CNN2 in colon cancer cell lines and determined the effects of this protein by increasing expression. METHODS: We used IHC and RT-PCR to measure CNN2 expression in colon cancer tissues and normal tissues and found increased expression levels in cancer tissues. We used viral vectors to decrease the level of CNN2 in the SW480 colon cancer cell line and found that silencing of CNN2 inhibited cell invasion. We detected potential protein interactions in signal pathways by western bolt analysis. To confirm the effect of CNN2 on cell invasion, we increased CNN2 expression in the SW620 cell line and observed increased invasion increased expression levels of associated proteins. RESULTS: High-expression levels of CNN2 were detected in cancer tissues. Silencing CNN2 inhibited cell invasion, down-regulated N-cadherin (N-CA) and C-myc proteins, and up-regulated E-cadherin (E-CA), suggesting that CNN2 promotes colon cancer cell invasion. Increasing CNN2 levels in the SW620 cell line showed the opposite results. CONCLUSIONS: CNN2 may act to promote colon cancer.
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Tissue-derived RNA, DNA and protein samples become more and more crucial for molecular detection in clinical research, personalized and targeted cancer therapy. This study evaluated how to biobanking colorectal tissues through examining the influences of cold ischemic time and freeze-thaw cycles on RNA, DNA and protein integrity. Here, 144 pairs of tumor and normal colorectal tissues were used to investigate the impact of cold ischemic times (0-48h) on RNA, DNA and protein integrity at on ice or room temperature conditions. Additionally, 45 pairs of tissues experienced 0-9 freeze-thaw cycles, and then the RNA, DNA and protein quality were analyzed. On ice, RNA, DNA and protein from colorectal tumor and normal tissues were all stable up to 48h after surgery. At room temperature, RNA in colorectal tumor and normal tissues began to degrade at 8h and 24h, respectively. Meanwhile, the tumor tissues DNA degradation occurred at 24h after surgery at room temperature. Similarly, the protein expression level of tumor and normal tissues began to change at 24h after the surgery at room temperature. Interestingly, tissue RNA and DNA remained stable even after 9 freeze-thaw cycles, whereas the proteins levels were remarkably changed after 7 freeze-thaw cycles. This study provided a useful evidence on how to store human colorectal tissues for biobanking. Preserving the surgical colorectal tissue on ice was an effective way to prevent RNA, DNA and protein degradation. Importantly, more than 7 repeated freeze-thaw cycles were not recommended for colorectal tissues.
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In the version of this article initially published, what was originally described as 'conditioned place preference' in a two-chamber mouse experiment could be better described as 'conditioned place avoidance'.
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Cervical radiculopathic pain is a very common symptom that may occur with cervical spondylosis. Mechanical allodynia is often associated with cervical radiculopathic pain and is inadequately treated with current therapies. However, the precise mechanisms underlying cervical radiculopathic pain-associated mechanical allodynia have remained elusive. Compelling evidence from animal models suggests a role of large-diameter dorsal root ganglion neurons and plasticity of spinal circuitry attached with Aß fibers in mediating neuropathic pain. Whether cervical radiculopathic pain condition induces plastic changes of large-diameter dorsal root ganglion neurons and what mechanisms underlie these changes are yet to be known. With combination of patch-clamp recording, immunohistochemical staining, as well as behavioral surveys, we demonstrated that upon chronic compression of C7/8 dorsal root ganglions, large-diameter cervical dorsal root ganglion neurons exhibited frequent spontaneous firing together with hyperexcitability. Quantitative analysis of hyperpolarization-activated cation current ( Ih) revealed that Ih was greatly upregulated in large dorsal root ganglion neurons from cervical radiculopathic pain rats. This increased Ih was supported by the enhanced expression of hyperpolarization-activated, cyclic nucleotide-modulated channels subunit 3 in large dorsal root ganglion neurons. Blockade of Ih with selective antagonist, ZD7288 was able to eliminate the mechanical allodynia associated with cervical radiculopathic pain. This study sheds new light on the functional plasticity of a specific subset of large-diameter dorsal root ganglion neurons and reveals a novel mechanism that could underlie the mechanical allodynia associated with cervical radiculopathy.
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Ganglios Espinales/citología , Ganglios Espinales/metabolismo , Neuralgia/etiología , Neuralgia/metabolismo , Neuronas/citología , Neuronas/metabolismo , Radiculopatía/etiología , Radiculopatía/metabolismo , Animales , Dolor Crónico/etiología , Dolor Crónico/metabolismo , Dolor Crónico/patología , Masculino , Potenciales de la Membrana/fisiología , Neuralgia/patología , Neuronas Aferentes/citología , Neuronas Aferentes/metabolismo , Radiculopatía/patología , Ratas , Ratas Sprague-DawleyRESUMEN
Programmed cell death ligand-1 (PD-L1) is typically produced by cancer cells and suppresses immunity through the receptor PD-1 expressed on T cells. However, the role of PD-L1 and PD-1 in regulating pain and neuronal function is unclear. Here we report that both melanoma and normal neural tissues including dorsal root ganglion (DRG) produce PD-L1 that can potently inhibit acute and chronic pain. Intraplantar injection of PD-L1 evoked analgesia in naive mice via PD-1, whereas PD-L1 neutralization or PD-1 blockade induced mechanical allodynia. Mice lacking Pd1 (Pdcd1) exhibited thermal and mechanical hypersensitivity. PD-1 activation in DRG nociceptive neurons by PD-L1 induced phosphorylation of the tyrosine phosphatase SHP-1, inhibited sodium channels and caused hyperpolarization through activation of TREK2 K+ channels. PD-L1 also potently suppressed nociceptive neuron excitability in human DRGs. Notably, blocking PD-L1 or PD-1 elicited spontaneous pain and allodynia in melanoma-bearing mice. Our findings identify a previously unrecognized role of PD-L1 as an endogenous pain inhibitor and a neuromodulator.
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Analgesia , Antígeno B7-H1/fisiología , Cultivo Primario de Células , Receptor de Muerte Celular Programada 1/fisiología , Animales , Antígeno B7-H1/sangre , Antígeno B7-H1/farmacología , Células Cultivadas , Cricetinae , Femenino , Ganglios Espinales/efectos de los fármacos , Ganglios Espinales/metabolismo , Ganglios Espinales/fisiología , Humanos , Hiperalgesia/inducido químicamente , Masculino , Melanoma/sangre , Melanoma/fisiopatología , Ratones , Ratones Noqueados , Neuralgia/fisiopatología , Neuronas/efectos de los fármacos , Neuronas/fisiología , Umbral del Dolor/efectos de los fármacos , Umbral del Dolor/fisiología , Fosforilación , Canales de Potasio de Dominio Poro en Tándem/fisiología , Receptor de Muerte Celular Programada 1/biosíntesis , Receptor de Muerte Celular Programada 1/genética , Proteína Tirosina Fosfatasa no Receptora Tipo 6/metabolismo , Ratas , Canales de Sodio/fisiología , Transmisión Sináptica/efectos de los fármacos , Transmisión Sináptica/fisiologíaRESUMEN
Relapse and metastasis are frequent in colon cancer and may be linked to stem cell characteristics. This study isolated side population (SP) cells from a colon cancer cell line (Colo-320) and examined their self-renewal and differentiation abilities. Compared to non-SP (NSP) cells, SP colon cancer cells were more tumorigenic in vivo and exhibited more invasive characteristics and a greater ability to form colonies. Additionally, more cells were in G0/G1 phase and more highly expressed the multidrug resistance protein BCRP/ABCG2. We achieved enhanced chemotherapy sensitivity by transfecting SP cells with a hairpin-like, small interfering RNA (siRNA) eukaryotic expression plasmid targeting BCRP/ABCG2.
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Transportador de Casetes de Unión a ATP, Subfamilia G, Miembro 2/antagonistas & inhibidores , Neoplasias del Colon/terapia , Fluorouracilo/administración & dosificación , Proteínas de Neoplasias/antagonistas & inhibidores , Células Madre Neoplásicas/efectos de los fármacos , ARN Interferente Pequeño/farmacología , Células de Población Lateral/efectos de los fármacos , Transportador de Casetes de Unión a ATP, Subfamilia G, Miembro 2/genética , Animales , Diferenciación Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Neoplasias del Colon/genética , Neoplasias del Colon/metabolismo , Sinergismo Farmacológico , Quimioterapia , Fluorouracilo/farmacología , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Ratones , Proteínas de Neoplasias/genética , Células Madre Neoplásicas/patología , Células de Población Lateral/patología , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Previous studies have shown that activation of p38 mitogen-activating kinase (MAPK) in spinal microglia participates in the generation of inflammatory and neuropathic pain in various rodent models. However, these studies focused on male mice to avoid confounding effects of the estrous cycle of females. Recent studies have shown that some spinal pro-inflammatory signaling such as Toll-like receptor 4-mediated signaling contributes to pain hypersensitivity only in male mice. In this study we investigated the distinct role of spinal p38 in inflammatory and neuropathic pain using a highly selective p38 inhibitor skepinone. Intrathecal injection of skepinone prevented formalin induced inflammatory pain in male but not female mice. Furthermore, intrathecal skepinone reduced chronic constriction injury (CCI) induced neuropathic pain (mechanical allodynia) in male mice on CCI-day 7 but not CCI-day 21. This male-dependent inhibition of neuropathic pain also occurred in rats following intrathecal skepinone. Nerve injury induced spinal p38 activation (phosphorylation) in CX3CR1-GFP(+) microglia on CCI-day 7, and this activation was more prominent in male mice. In contrast, CCI induced comparable microgliosis and expression of the microglial markers CX3CR1 and IBA-1 in both sexes. Notably, intraperitoneal or local perineural administration of skepinone inhibited CCI-induced mechanical allodynia in both sexes of mice. Finally, skepinone only reduced the frequency of spontaneous excitatory postsynaptic currents (sEPSCs) in lamina IIo neurons of spinal cord slices of males 7days post CCI. Therefore, the sex-specific p38 activation and signaling is confined to the spinal cord in inflammatory and neuropathic pain conditions.
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Inflamación/tratamiento farmacológico , Microglía/metabolismo , Neuralgia/tratamiento farmacológico , Inhibidores de Proteínas Quinasas/farmacología , Médula Espinal/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/antagonistas & inhibidores , Animales , Dibenzocicloheptenos/administración & dosificación , Dibenzocicloheptenos/farmacología , Modelos Animales de Enfermedad , Femenino , Masculino , Ratones , Ratones Endogámicos C57BL , Inhibidores de Proteínas Quinasas/administración & dosificación , Factores SexualesRESUMEN
Cervical radiculopathy represents aberrant mechanical hypersensitivity. Primary sensory neuron's ability to sense mechanical force forms mechanotransduction. However, whether this property undergoes activity-dependent plastic changes and underlies mechanical hypersensitivity associated with cervical radiculopathic pain (CRP) is not clear. Here we show a new CRP model producing stable mechanical compression of dorsal root ganglion (DRG), which induces dramatic behavioral mechanical hypersensitivity. Amongst nociceptive DRG neurons, a mechanically sensitive neuron, isolectin B4 negative Aδ-type (IB4(-) Aδ) DRG neuron displays spontaneous activity with hyperexcitability after chronic compression of cervical DRGs. Focal mechanical stimulation on somata of IB4(-) Aδ neuron induces abnormal hypersensitivity. Upregulated HCN1 and HCN3 channels and increased Ih current on this subset of primary nociceptors underlies the spontaneous activity together with neuronal mechanical hypersensitivity, which further contributes to the behavioral mechanical hypersensitivity associated with CRP. This study sheds new light on the functional plasticity of a specific subset of nociceptive DRG neurons to mechanical stimulation and reveals a novel mechanism that could underlie the mechanical hypersensitivity associated with cervical radiculopathy.
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Ganglios Espinales/metabolismo , Ganglios Espinales/fisiopatología , Canales Regulados por Nucleótidos Cíclicos Activados por Hiperpolarización/genética , Mecanotransducción Celular , Nociceptores/metabolismo , Radiculopatía/genética , Radiculopatía/fisiopatología , Animales , Modelos Animales de Enfermedad , Expresión Génica , Genes fos , Hiperalgesia/etiología , Potenciales de la Membrana , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , Fibras Nerviosas Mielínicas/metabolismo , Neuralgia/etiología , Fosforilación , Radiculopatía/complicaciones , Radiculopatía/etiología , Ratas , Regulación hacia ArribaRESUMEN
Adjuvant interferon (IFN) therapy following curative treatment for hepatocellular carcinoma (HCC) has been extensively investigated; however, the clinical benefits with different hepatitis backgrounds remain unclear. Medline, Embase, PubMed and the Cochrane Library databases were searched to identify randomized trials and cohort studies that enrolled HCC patients who received curative surgery or ablation therapy followed by IFN and control subjects; the studies were required to include data on early or late recurrence and mortality rates of HCC. Hepatitis B virus (HBV) associated with HCC (HBV-HCC) and hepatitis C virus (HCV) associated with HCC (HCV-HCC) were separately analyzed and recurrence, mortality and clinicopathological factors were compared. A total of 14 studies (9 randomized trials and 5 cohort studies, including 1,385 patients in total) were eligible for meta-analysis. IFN was found to decrease mortality and early recurrence rates, but exerted no effect on late recurrence rate. The effect of IFN differed between HBV-HCC and HCV-HCC cases. In HCV-HCC, IFN significantly reduced mortality as well as recurrence rates. However, in HBV-HCC patients, IFN reduced mortality rather than recurrence rates, although it also reduced the recurrence rate in certain subgroups. In conclusion, the effect of adjuvant IFN on postoperative recurrence differed between HBV-HCC and HCV-HCC cases; therefore, different strategies with adjuvant IFN should be used to treat HCC with different hepatitis backgrounds.
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BACKGROUND:There is no unified method for treatment of intertrochanteric fracture. Previous dynamic hip screw fixation has some shortcomings, and is gradual y replaced by intramedul ary nail fixation. interTan is a compression interlocking intramedul ary nail designed by Smith Nephew in USA according the characteristics of intertrochanteric fracture. Unique design of pressurized screw thread has control able linear pressure. Moreover, it has the stability of Z-effect anti-rotation. OBJECTIVE:To analyze the clinical therapeutic effects and superiority of interTan compression interlocking intramedul ary nail for intertrochanteric fracture. METHODS:A total of 60 patients with intertrochanteric fracture undergoing interTan compression interlocking intramedul ary nail in the Department of Orthopedics, Beijing Chaoyang Emergency Rescue Center from January 2011 to December 2012 were enrol ed in the experimental group. A total of 60 patients with intertrochanteric fractures receiving dynamic hip screw fixation in the Department of Orthopedics, Beijing Chaoyang Emergency Rescue Center from January 2009 to December 2010 were enrol ed in the control group. RESULTS AND CONCLUSION:Peri-operative bleeding volume, postoperative ambulation time, numerical rating scale score, radiographic healing time and weight-bearing time in the experimental group were lower than those of control group. At 3, 6 months and 1 year after fixation, Harris score and hospital for special surgery knee score were higher in the experimental group than in the control group. These data indicated that compared with dynamic hip screw, interTan compression interlocking intramedul ary nail has unique stability due to its design. Internal fixation can reduce operation trauma, promote fracture healing and improve joint function, and exhibit good clinical therapeutic effects for intertrochanteric fractures.
RESUMEN
Synaptic long-term potentiation (LTP) at spinal neurons directly communicating pain-specific inputs from the periphery to the brain has been proposed to serve as a trigger for pain hypersensitivity in pathological states. Previous studies have functionally implicated the NMDA receptor-NO pathway and the downstream second messenger, cGMP, in these processes. Because cGMP can broadly influence diverse ion-channels, kinases, and phosphodiesterases, pre- as well as post-synaptically, the precise identity of cGMP targets mediating spinal LTP, their mechanisms of action, and their locus in the spinal circuitry are still unclear. Here, we found that Protein Kinase G1 (PKG-I) localized presynaptically in nociceptor terminals plays an essential role in the expression of spinal LTP. Using the Cre-lox P system, we generated nociceptor-specific knockout mice lacking PKG-I specifically in presynaptic terminals of nociceptors in the spinal cord, but not in post-synaptic neurons or elsewhere (SNS-PKG-I(-/-) mice). Patch clamp recordings showed that activity-induced LTP at identified synapses between nociceptors and spinal neurons projecting to the periaqueductal grey (PAG) was completely abolished in SNS-PKG-I(-/-) mice, although basal synaptic transmission was not affected. Analyses of synaptic failure rates and paired-pulse ratios indicated a role for presynaptic PKG-I in regulating the probability of neurotransmitter release. Inositol 1,4,5-triphosphate receptor 1 and myosin light chain kinase were recruited as key phosphorylation targets of presynaptic PKG-I in nociceptive neurons. Finally, behavioural analyses in vivo showed marked defects in SNS-PKG-I(-/-) mice in several models of activity-induced nociceptive hypersensitivity, and pharmacological studies identified a clear contribution of PKG-I expressed in spinal terminals of nociceptors. Our results thus indicate that presynaptic mechanisms involving an increase in release probability from nociceptors are operational in the expression of synaptic LTP on spinal-PAG projection neurons and that PKG-I localized in presynaptic nociceptor terminals plays an essential role in this process to regulate pain sensitivity.
Asunto(s)
Proteínas Quinasas Dependientes de GMP Cíclico/metabolismo , Potenciación a Largo Plazo , Nociceptores/metabolismo , Dolor/patología , Aminoquinolinas/farmacología , Animales , Conducta Animal , Moléculas de Adhesión Celular/genética , Moléculas de Adhesión Celular/metabolismo , Proteína Quinasa Dependiente de GMP Cíclico Tipo I , Proteínas Quinasas Dependientes de GMP Cíclico/genética , Activación Enzimática , Ganglios Espinales/metabolismo , Ganglios Espinales/patología , Eliminación de Gen , Guanilato Ciclasa/antagonistas & inhibidores , Guanilato Ciclasa/metabolismo , Receptores de Inositol 1,4,5-Trifosfato/genética , Receptores de Inositol 1,4,5-Trifosfato/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Proteínas de Microfilamentos/genética , Proteínas de Microfilamentos/metabolismo , Fibras Nerviosas/metabolismo , Fibras Nerviosas/patología , Nociceptores/efectos de los fármacos , Nociceptores/patología , Dolor/metabolismo , Técnicas de Placa-Clamp , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Fosforilación , Transducción de Señal , Especificidad por Sustrato , Transmisión SinápticaRESUMEN
The transient receptor potential vanilloid subfamily member 1 (TRPV1) is a protein mainly expressed in sensory neurons and fibers, such as in trigeminal ganglion and dorsal root ganglion, and has been indicated to be involved in several physiological and pathological processes. Studies on thermal activation have revealed that phosphorylation is involved in TRPV1 activation and 2 putative phosphorylation sites, Ser residues 502 (Ser-502) and Ser residues 800 (Ser-800), have been recently confirmed to possess the capability of resensitizing TRPV1. In addition to acidification, alkalization has also been proved to be a highly effective stimulator for TRPV1. TRPV1 could be regulated by various physical and chemical modulators, as well as the chronic pain. TRPV1 plays a crucial role in the transmission of pain signals, especially under inflammation and the neoplasm conditions, and it can also modulate nociceptive afferents by reinforcing morphine tolerance. The present review mainly focused on the structural and functional complexities of TRPV1, together with its activation and modulation by a wide variety of physical and chemical stimuli. Its pharmacological manipulation (sensitization/desensitization) and therapeutical targets were also discussed.