RESUMEN

Objective: This study aimed to investigate the diagnostic value of a combination of anti-extractable nuclear antigens (anti-ENA) antibodies, anti-cardiolipin antibodies (ACA), and anti-ß2-glycoprotein 1 (anti-ß2 GPI) antibodies in patients with systemic lupus erythematosus (SLE). Methods: A total of 646 SLE patients diagnosed in our hospital between January 2020 and April 2023 were randomly selected as study subjects, while 2075 non-SLE subjects during the same period were selected as the control group. The levels of anti-extractable nuclear antigen (ENA) antibody, ACA, and anti-ß2 GPI antibodies were measured, and their diagnostic value for SLE was analyzed using binary logistic regression and receiver operating characteristic (ROC) analysis. Results: The rates of positive anti-RNP, anti-Sm, anti-Sjögren's syndrome (SS-A), and anti-SS-B antibodies in the SLE patient group were significantly higher than those in the control group, with all differences being statistically significant (P < 0.01). The rates of positive ACA, as well as the levels of ACA IgA, ACA IgG, and ACA IgM, were significantly higher in the SLE patients group compared to the control group, with statistically differences (P < 0.05). Similarly, the rates of positive anti-ß2 GPI antibodies, anti-ß2 GPI antibody IgA, IgG, and IgM, were significantly higher in the SLE patient group compared to the control group, with all differences being statistically significant (P < 0.01). Gender, age, positive anti-JO1 antibodies, positive anti-RNP antibodies, positive anti-SS-A antibodies, positive ACA, high level ACA IgG and ACA IgM, positive anti-ß2 GPI antibodies, and high level of anti-ß2 GPI antibody IgA were identified as independent risk factors for the development of SLE (P < 0.05). The combined use of age, sex, anti-ENA antibodies, ACA, and anti-ß2 GPI antibodies yielded a sensitivity of 82.12% (80.41%-83.71%) and a specificity of 80.03% (76.77%-82.93%) for the diagnosis of SLE. Conclusion: The combination of age, sex, anti-ENA antibodies, ACA, and anti-ß2 GPI antibodies shows high diagnostic value for SLE and holds potential for clinical application as an auxiliary diagnostic tool for SLE.

RESUMEN

It has been verified that, as an emerging contaminant, microplastics are capable of adsorbing certain traditional contaminants like the heavy metal Cd. However, the majority of previous studies only focused on certain types of virgin microplastics, especially for PE and PS. In addition, this adsorption process might be affected by microplastics inevitably undergoing aging and consequent changes in the natural environment. Unfortunately, the relevant reports on aging effects were mainly about organic pollutants, rather than heavy metals. By far, there have been few comprehensive and mechanistic studies on the key aging effects on the Cd adsorption by various types of microplastics. In this study, five representative types of microplastics (i.e., PS, ABS, PP, PVC, and PET) were selected for aging by ultraviolet radiation, and the physicochemical properties of virgin and aged microplastics were thoroughly compared, including specific surface area, crystallinity, surface functional groups, and surface elements. Accordingly, the changes in adsorption isotherms of Cd by microplastics were discussed. The results showed that:① aging induced non-significant changes in specific surface area but a significant decrease in crystallinity. Surface functional groups also changed, including the emergence of a C=O functional group on PS and ABS, the decrease in C=C absorption peak intensity on ABS, and the increase in absorption peak intensities of C=O, C-O, and polar ester groups on PET. Regarding surface C content, C=C/C-C decreased, whereas C-O and O-C=O increased. The total O content and O/C significantly increased as well. ② The Langmuir model well-fitted the adsorption isotherms of Cd by virgin and aged microplastics. Aging significantly expanded the adsorption capacity of Cd by microplastics, as the order of saturated adsorption capacity before aging was ABS (0.2284 mg·g-1)>PVC (0.1360 mg·g-1)>PS (0.1286 mg·g-1)>PP (0.1005 mg·g-1)>PET (0.0462 mg·g-1) and then became PS (0.2768 mg·g-1)>ABS (0.2586 mg·g-1)>PVC (0.1776 mg·g-1)>PP (0.1721 mg·g-1)>PET (0.0951 mg·g-1) after aging. ③ Both crystallinity and surface functional groups played key roles in the adsorption of Cd by microplastics. As for virgin microplastics, crystallinity was negatively correlated with the saturated adsorption capacity of Cd, because the amorphous regions contributed most to Cd adsorption. Aging brought about the decrease in crystallinity and the increase in amorphous regions, which further promoted the oxidation reaction on microplastics. Consequently, oxygen-containing functional groups increased on the surface and eventually expanded the adsorption capacity of Cd by microplastics. Note that certain specific functional groups of various microplastics also had impacts on the adsorption process. These results provide valuable information about the environmental behaviors and interactions of microplastics and heavy metals in nature.

Asunto(s)

Metales Pesados , Contaminantes Químicos del Agua , Adsorción , Cadmio , Microplásticos , Plásticos/química , Cloruro de Polivinilo , Rayos Ultravioleta , Contaminantes Químicos del Agua/análisis

RESUMEN

Sunitinib is the most commonly used first-line therapy for the treatment of advanced renal cell carcinoma (RCC), but intrinsic and extrinsic resistance to targeted therapies dramatically compromise the benefit of clinical outcome. Dissecting the underlying mechanisms and discovering reliable predictive biomarkers are urgently needed in clinic. Here, we discovered miR-885-5p was notably decreased after sunitinib treatment and associated with poor disease progression in clear cell renal cell carcinoma (ccRCC). In vitro and in vivo studies identified miR-885-5p inhibition contributed to sunitinib resistance. Mechanistically, sunitinib treatment reduced GATA1 expression, which in turn reduced its binding to MIR885 promoter and resulted in miR-885-5p downregulation in transcriptional level. In addition, PLIN3 was confirmed to be directly targeted by miR-885-5p and its upregulation significantly increased lipid droplets formation to decrease sunitinib sensitivity. Therefore, GATA1/miR-885-5p/ PLIN3 pathway may serve as a potential therapeutic strategy and a biomarker for sunitinib treatment in ccRCC.

Asunto(s)

Carcinoma de Células Renales/tratamiento farmacológico , Retroalimentación Fisiológica , Factor de Transcripción GATA1/metabolismo , Neoplasias Renales/tratamiento farmacológico , MicroARNs/metabolismo , Perilipina-3/metabolismo , Sunitinib/uso terapéutico , Animales , Secuencia de Bases , Carcinoma de Células Renales/genética , Carcinoma de Células Renales/patología , Línea Celular Tumoral , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/genética , Resistencia a Antineoplásicos/efectos de los fármacos , Resistencia a Antineoplásicos/genética , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Neoplasias Renales/patología , Gotas Lipídicas/metabolismo , Masculino , Ratones Endogámicos BALB C , Ratones Desnudos , MicroARNs/genética , Persona de Mediana Edad , Regiones Promotoras Genéticas/genética , Transducción de Señal/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/genética

RESUMEN

On the basis and continuation of our previous studies on anti-tubulin and anti-gastric cancer agents, novel tertiary amide derivatives incorporating benzothiazole moiety were synthesized and the antiproliferative activity was studied in vitro. Preliminary structure activity relationships (SARs) were explored according to the in vitro antiproliferative activity results. Some of compounds could significantly inhibit the proliferation of three cancer cells (HCT-116, MGC-803 and PC-3 cells) and compound F10 exhibited excellent antiproliferative activity against HCT-116 cells (IC50 = 0.182 µM), MGC-803 cells (IC50 = 0.035 µM), PC-3 cells(IC50 = 2.11 µM) and SGC-7901 cells (IC50 = 0.049 µM). Compound F10 effectively inhibited tubulin polymerization (IC50 = 1.9 µM) and bound to colchicine binding site of tubulin. Molecular docking results suggested compound F10 could bind tightly into the colchicine binding site of ß-tubulin. Moreover, compound F10 could regulate the Hippo/YAP signaling pathway. Compound F10 activated Hippo signaling pathway from its very beginning MST1/2, as the result of Hippo cascade activation YAP were inhibited. And then it led to a decrease of c-Myc and Bcl-2 expression. Further molecular experiments showed that compound F10 arrested at G2/M phase, inhibited cell colony formatting and induced extrinsic and intrinsic apoptosis in MGC-803 and SGC-7901 cells. Collectively, compound F10 was the first to be reported as a new anticancer agent in vitro via inhibiting tubulin polymerization and activating the Hippo signaling pathway.

Asunto(s)

Amidas/química , Benzotiazoles/química , Benzotiazoles/farmacología , Proteínas Serina-Treonina Quinasas/metabolismo , Transducción de Señal/efectos de los fármacos , Neoplasias Gástricas/patología , Tubulina (Proteína)/química , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Antineoplásicos/química , Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Diseño de Fármacos , Puntos de Control de la Fase G2 del Ciclo Celular/efectos de los fármacos , Vía de Señalización Hippo , Humanos , Puntos de Control de la Fase M del Ciclo Celular/efectos de los fármacos , Multimerización de Proteína/efectos de los fármacos , Estructura Cuaternaria de Proteína , Factores de Transcripción/metabolismo , Proteínas Señalizadoras YAP

RESUMEN

BACKGROUND: Prostate cancer (PCa) is one of the commonest male urinary and reproductive system malignancies with high morbidity and mortality. circLMTK2 was reported as a tumor suppressor, therefore, we attempted to investigate the potential mechanism of circLMTK2 in PCa. METHODS: qRT-PCR was employed to examine the expressions of circLMTK2 and miR-183. Afterwards, cell transfection was conducted for overexpressing circLMTK2 and miR-183 in LNCaP and PC3 cells, and silencing circLMTK2 in RWPE1 cells. Then, CCK-8 assay, BrdU, transwell assay, flow cytometry and western blot were respectively conducted to examine the variations of cell growth and metastasis, as well as apoptosis. The expressions of key proteins involved in Wnt/ß-catenin and PI3K/AKT pathways were further investigated utilizing western blot. RESULTS: circLMTK2 was lowly expressed in tumor tissues. circLMTK2 overexpression suppressed cell proliferation and metastasis, however promoted cell apoptosis in LNCaP and PC3 cells. circLMTK2 knockdown enhanced cell viability, proliferation, migration and invasion, while had no significant influences on apoptosis of RWPE1 cells. Further experiments verified that miR-183 up-regulation counteracted the influences triggered by circLMTK2 overexpression in LNCaP and PC3 cells. Besides, it markedly promoted the viability, proliferation, migration and invasion of LNCaP cells, however had no significant influence on cell apoptosis. Moreover, the inhibitory effects on Wnt/ß-catenin and PI3K/AKT pathways evoked by circLMTK2 overexpression were diminished by miR-183 up-regulation in LNCaP and PC3 cells. CONCLUSION: These outcomes illustrated that circLMTK2 overexpression exerts an anti-tumor effects through down-regulating the expression of miR-183.

Asunto(s)

Genes Supresores de Tumor , MicroARNs/genética , Neoplasias de la Próstata/genética , ARN Circular/genética , Anciano , Apoptosis/genética , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Regulación Neoplásica de la Expresión Génica , Silenciador del Gen , Humanos , Masculino , Proteínas de la Membrana/genética , Persona de Mediana Edad , Fosfatidilinositol 3-Quinasas/genética , Fosfatidilinositol 3-Quinasas/metabolismo , Neoplasias de la Próstata/patología , Proteínas Serina-Treonina Quinasas/genética , Proteínas Proto-Oncogénicas c-akt/genética , Proteínas Proto-Oncogénicas c-akt/metabolismo , Vía de Señalización Wnt/genética

RESUMEN

Background: Prostate cancer (PC) fearfully impacts men's health. We explored the efficacy and mechanism of circular RNA circZNF609 (circZNF609) on colony formation, viability, apoptosis, migration and invasion and in PC cells. Methods: Colony formation, CCK-8, flow cytometry, migration and invasion assay were respectively used to detect the functions of circZNF609 and microRNA (miR)-186-5p on cell colony ability, viability, apoptosis, migration and invasion. circZNF609 and miR-186-5p expression were changed by cell transfection and tested by RT-qPCR. Moreover, Cleaved-Caspase-3, Cleaved-Caspase-9, matrix metalloproteinase-9 (MMP-9), Vimentin and relate-proteins of cell pathways were examined through Western blot. Results: circZNF609 was highly expressed at PC tissues. circZNF609 declined cell colony ability, viability, migration and invasion and caused apoptosis. Furthermore, circZNF609 negatively regulated miR-186-5p, miR-186-5p inhibitor could reverse impacts of circZNF609. Finally, circZNF609 restrained the YAP1 and AMPK pathways by up-regulating miR-186-5p. Conclusion: Silencing circZNF609 restrained growth, migration and invasion of PC cells by up-regulating miR-186-5p via YAP1 and AMPK pathways. Highlights circZNF609 is highly expressed in PC tissues; circZNF609 restrains cell growth, migration and invasion; circZNF609 exerts its function by up-regulating miR-186-5p; circZNF609 exerts its function by YAP1 and AMPK signaling pathways.

Asunto(s)

Movimiento Celular/genética , MicroARNs/genética , Neoplasias de la Próstata/patología , Interferencia de ARN , ARN Circular/deficiencia , ARN Circular/genética , Regulación hacia Arriba/genética , Proteínas Quinasas Activadas por AMP/metabolismo , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Proliferación Celular/genética , Humanos , Masculino , Invasividad Neoplásica/genética , Células PC-3 , Neoplasias de la Próstata/genética , Transducción de Señal/genética , Factores de Transcripción/metabolismo , Proteínas Señalizadoras YAP

RESUMEN

The aim of the present study was to investigate the effect and therapeutic potential of baicalin in breast cancer. Baicalin is used to treat inflammatory diseases. The effects of baicalin were assessed in breast cancer MCF-7 and MDA-MB­231 cells, and human breast cancer xenograft mice. Cells were treated with 0, 20 or 30 µM baicalin for 48 h, while xenograft mice were treated with intraperitoneal injection of 0, 100 or 200 mg/kg baicalin for 30 days. The results demonstrated that treatment with baicalin dose-dependently suppressed breast cancer cell invasion, migration and proliferation, and also induced G1/S-phase cell cycle arrest in vitro and in vivo. Baicalin alleviated inflammation injury and inhibited the secretion of tumor necrosis factor (TNF)-α and interleukin (IL)-1ß, thus suppressing nuclear factor (NF)-ĸB-p65 activation via inhibition of IĸB kinase. Investigation of the mechanism underlying baicalin activity indicated that it inhibited protein expression of NF-ĸB-p65, leading to NF-ĸB­induced increased expression of CCND1, BCL2, BIRC2 and BIRC3, thus inhibiting cell proliferation, invasion and migration and suppressing anti-apoptotic factors in vitro and in vivo. In addition, baicalin did not affect non-tumorigenic normal breast epithelial cells. These results indicate that baicalin may exert therapeutic effects in breast cancer.

Asunto(s)

Neoplasias de la Mama/tratamiento farmacológico , Flavonoides/administración & dosificación , Quinasa I-kappa B/metabolismo , Animales , Neoplasias de la Mama/metabolismo , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Esquema de Medicación , Femenino , Flavonoides/farmacología , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Inyecciones Intraperitoneales , Células MCF-7 , Ratones , Transducción de Señal/efectos de los fármacos , Ensayos Antitumor por Modelo de Xenoinjerto

RESUMEN

OBJECTIVE: Results of in vitro and in vivo studies suggest that consumption of beer is less harmful for the liver than consumption of spirits. It also has been suggested that secondary plant compounds derived from hops such as xanthohumol or iso-α-acids may have beneficial effects on the development of liver diseases of various etiologies. The aim of this study was to determine whether iso-α-acids consumed in doses achieved by "normal" beer consumption have beneficial effects on health. METHODS: Female C57 Bl/6 J mice, pretreated for 4 d with an iso-α-acid-rich extract (∼30% iso-α-acids from hops, 0.75 mg/kg body weight), were fed one bolus of ethanol (6 g/kg body weight intragastric) or an iso-caloric maltodextrin solution. Markers of liver damage, toll-like receptor-4 signaling, and lipid peroxidation were determined. Furthermore, the effect of isohumulone on the lipopolysaccharide-dependent activation of J774 A.1 macrophages, used as a model of Kupffer cells, was determined. RESULTS: In the liver, acute ethanol administration led to a significant accumulation of fat (∼10-fold), which was accompanied by significantly higher inducible nitric oxide synthase protein level, elevated nitric oxide production, and increased plasminogen activator inhibitor 1 protein concentration when compared to controls. In mice pretreated with iso-α-acids, these effects of alcohol were markedly attenuated. Pretreatment of J774 A.1 macrophages with isohumulone significantly attenuated lipopolysaccharide-induced mRNA expression of inducible nitric oxide synthase and interleukin-6 as well as the release of nitric oxide. CONCLUSION: Taken together, iso-α-acids markedly attenuated the development of acute alcohol-induced damage in mice.

Asunto(s)

Ácidos/farmacología , Hígado Graso Alcohólico/tratamiento farmacológico , Humulus/química , Extractos Vegetales/farmacología , Animales , Línea Celular , Femenino , Interleucina-6/genética , Interleucina-6/metabolismo , Macrófagos del Hígado/efectos de los fármacos , Macrófagos del Hígado/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Lipopolisacáridos/efectos adversos , Hígado/efectos de los fármacos , Hígado/metabolismo , Ratones , Ratones Endogámicos C57BL , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Inhibidor 1 de Activador Plasminogénico/genética , Inhibidor 1 de Activador Plasminogénico/metabolismo , Receptor Toll-Like 4/genética , Receptor Toll-Like 4/metabolismo

RESUMEN

Ternary semiconductor nanowire arrays enable scalable fabrication of nano-optoelectronic devices with tunable bandgap. However, the lack of insight into the effects of the incorporation of Vy element results in lack of control on the growth of ternary III-V(1-y)Vy nanowires and hinders the development of high-performance nanowire devices based on such ternaries. Here, we report on the origins of Sb-induced effects affecting the morphology and crystal structure of self-catalyzed GaAsSb nanowire arrays. The nanowire growth by molecular beam epitaxy is changed both kinetically and thermodynamically by the introduction of Sb. An anomalous decrease of the axial growth rate with increased Sb2 flux is found to be due to both the indirect kinetic influence via the Ga adatom diffusion induced catalyst geometry evolution and the direct composition modulation. From the fundamental growth analyses and the crystal phase evolution mechanism proposed in this Letter, the phase transition/stability in catalyst-assisted ternary III-V-V nanowire growth can be well explained. Wavelength tunability with good homogeneity of the optical emission from the self-catalyzed GaAsSb nanowire arrays with high crystal phase purity is demonstrated by only adjusting the Sb2 flux.

RESUMEN

Using a scanning tunnelling microscope break-junction technique, we produce 4,4'-bipyridine (44BP) single-molecule junctions with Ni and Au contacts. Electrochemical control is used to prevent Ni oxidation and to modulate the conductance of the devices via nonredox gating--the first time this has been shown using non-Au contacts. Remarkably the conductance and gain of the resulting Ni-44BP-Ni electrochemical transistors is significantly higher than analogous Au-based devices. Ab-initio calculations reveal that this behavior arises because charge transport is mediated by spin-polarized Ni d-electrons, which hybridize strongly with molecular orbitals to form a "spinterface". Our results highlight the important role of the contact material for single-molecule devices and show that it can be varied to provide control of charge and spin transport.

Asunto(s)

Oro , Transistores Electrónicos , Níquel , Piridinas

RESUMEN

There is increasing evidence suggesting that dysregulation of some microRNAs (miRNAs) may contribute to tumor progression and metastasis and have been proposed to be key regulators of diverse biological processes such as transcriptional regulation, cell growth and tumorigenesis. Previous studies have shown that miR-137 is dysregulated in some malignancies, but its role in bladder cancer is still unknown. In our study, we find that miR-137 is up-regulated in human bladder cancer tissues and cell lines. Moreover, the higher level of miR-137 was associated with pM or pTNM stage in clinical bladder cancer patients. Enforced expression of miR-137 in bladder cancer cells significantly enhanced their proliferation, migration and invasion. Bioinformatics analysis identified the tumor suppressor gene PAQR3 as a potential miR-137 target gene. Further studies indicated that miR-137 suppressed the expression of PAQR3 by binding to its 3'-untranslated region. Silencing of PAQR3 by small interfering RNAs phenocopied the effects of miR-137 overexpression, whereas restoration of PAQR3 in bladder cancer cells bladder cancer cells overexpressing miR-137, partially reversed the suppressive effects of miR-137. These findings indicate that miR-137 could be a potential oncogene in bladder cancer.

Asunto(s)

Péptidos y Proteínas de Señalización Intracelular/genética , Proteínas de la Membrana/genética , MicroARNs/genética , Regulación hacia Arriba/genética , Neoplasias de la Vejiga Urinaria/genética , Neoplasias de la Vejiga Urinaria/patología , Secuencia de Bases , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular , Humanos , Invasividad Neoplásica/genética

RESUMEN

We study the effect of functional groups (CH3*4, OCH3, CH3, Cl, CN, F*4) on the electronic transport properties of 1,4-benzenediamine molecular junctions using the non-equilibrium Green function method. Exchange and correlation effects are included at various levels of theory, namely density functional theory (DFT), energy level-corrected DFT (DFT+Σ), Hartree-Fock and the many-body GW approximation. All methods reproduce the expected trends for the energy of the frontier orbitals according to the electron donating or withdrawing character of the substituent group. However, only the GW method predicts the correct ordering of the conductance amongst the molecules. The absolute GW (DFT) conductance is within a factor of two (three) of the experimental values. Correcting the DFT orbital energies by a simple physically motivated scissors operator, Σ, can bring the DFT conductances close to experiments, but does not improve on the relative ordering. We ascribe this to a too strong pinning of the molecular energy levels to the metal Fermi level by DFT which suppresses the variation in orbital energy with functional group.

RESUMEN

BACKGROUND: Stem cell therapy is a promising therapeutic strategy for stress urinary incontinence (SUI). However, its current efficacy is insufficient. OBJECTIVE: We designed a stem cell transplantation system that contains autologous adipose-derived stem cells (ADSC) and controlled-release nerve growth factor (NGF). We evaluated whether this system could enhance the therapeutic efficacy of ADSCs by periurethral coinjection in SUI rats. DESIGN, SETTING, AND PARTICIPANTS: We first tested for the presence of NGF receptors in rat ADSCs and observed the effect of NGF on ADSCs in vitro and in vivo. NGF was encapsulated within poly(lactic-co-glycolic acid-PLGA) microspheres (PLGA/NGF) to control its release. SUI was created in rats, and ADSCs were harvested, cultured from fat tissue, and retained for later transplantation. SUI rats then received different forms of periurethral injection therapy. Their urodynamic index was monitored. Eight weeks after injection, the SUI rats were sacrificed and their urethra removed for histologic evaluation. INTERVENTION: Forty SUI rats were allocated to five groups for receiving periurethral injection with phosphate-buffered saline (PBS), ADSC, ADSC+PLGA, ADSC+NGF, or ADSC+PLGA/NGF. Bladder capacities, abdominal leak point pressure (ALPP), and retrograde urethral perfusion pressure (RUPP) were reassessed at 2, 6, and 8 wk after injection. MEASUREMENTS: The rat SUI model was generated by bilateral pudendal nerve transection (PNT). Real-time polymerase chain reaction (RT-PCR) and western blotting detected the NGF receptor Ark-A. The regeneration of muscles and peripheral nerves was evaluated by Masson's trichrome and immunohistochemical staining. RESULTS AND LIMITATIONS: Results revealed the presence of the NGF receptor Trk-A on rat ADSCs. Short-term observations showed that NGF could improve ADSCs' viability in vitro and in vivo. ADSCs delivered intramuscularly into the urethra in combination with PLGA/NGF resulted in significant improvements in ALPP and RUPP as well as the amount of muscle and ganglia. There was a significant difference between the ADSC+PLGA/NGF group and other groups. CONCLUSIONS: Periurethral coinjection of autologous ADSCs with controlled-release NGF may be a potential strategy for SUI treatment.

Asunto(s)

Tejido Adiposo/citología , Factor de Crecimiento Nervioso/administración & dosificación , Trasplante de Células Madre , Células Madre/efectos de los fármacos , Uretra/efectos de los fármacos , Incontinencia Urinaria de Esfuerzo/terapia , Animales , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Preparaciones de Acción Retardada , Modelos Animales de Enfermedad , Portadores de Fármacos , Femenino , Inyecciones , Ácido Láctico/química , Microesferas , Factor de Crecimiento Nervioso/química , Ácido Poliglicólico/química , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Ratas , Ratas Sprague-Dawley , Receptor trkA/agonistas , Receptor trkA/genética , Receptor trkA/metabolismo , Células Madre/metabolismo , Factores de Tiempo , Trasplante Autólogo , Uretra/patología , Uretra/fisiopatología , Incontinencia Urinaria de Esfuerzo/patología , Incontinencia Urinaria de Esfuerzo/fisiopatología , Urodinámica/efectos de los fármacos

RESUMEN

OBJECTIVES: RNA interference (RNAi) has the potential to be developed into therapeutics for prostate cancer, but the lack of cellular targets limits its application. In the present study we attempt to develop a prostate cancer-specific RNAi system using the human prostate specific membrane antigen (PSMA) promoter/enhancer; furthermore, we analyzed its inhibitive effect on STAT3 expression. METHODS: The adenoviral vectors containing a small hairpin RNA (shRNA) to target exogenous reporters enhance green fluorescent protein (EGFP) and endogenous gene signal transducers and activators of transcription 3 (STAT3) were constructed. After prostate cancer and other cells were transfected, reverse transcription-polymerase chain reaction (RT-PCR), fluorescence microscopy, and Western blotting were used to measure EGFP expression. Inhibition of STAT3 was evaluated by Western blotting. Cell proliferation and viability were measured by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Cell apoptosis was analyzed with double-staining of Annexin V and PI. RESULTS: Our study showed that with the PSMA promoter/enhancer directly driving shRNA transcription, expression of the exogenous reporters EGFP in prostate cancer cells, but not other cancer cells and normal cells, was specifically inhibited in vitro. The PSMA promoter/enhancer-driven shRNA also depressed the expression of STAT3 in only prostate cancer cells. Inhibition of STAT3 suppressed proliferation of PC-3 and LNCaP cells. CONCLUSIONS: The present study describes an efficient RNAi system for gene silencing that is specific to prostate cancer cells using the PSMA promoter/enhancer. Suppression of STAT3 by using this system decreased proliferation and induced apoptosis of PC-3 and LNCaP cells. This system may be useful for RNAi therapy for prostate cancer.

Asunto(s)

Antígenos de Superficie/genética , Regulación Neoplásica de la Expresión Génica/genética , Terapia Genética/métodos , Glutamato Carboxipeptidasa II/genética , Neoplasias de la Próstata/genética , Interferencia de ARN , Apoptosis/genética , Secuencia de Bases , Western Blotting , Línea Celular Tumoral , Proliferación Celular , Elementos de Facilitación Genéticos , Expresión Génica , Humanos , Masculino , Microscopía Fluorescente , Datos de Secuencia Molecular , Regiones Promotoras Genéticas/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Transcripción STAT3/biosíntesis , Transfección