We introduce a simple, versatile and robust one-step technique that enables real-time imaging of multiple intracellular caspase activities in living cells without the need for complicated synthetic protocols. Conventional fluorogenic probes or recently reported activatable probes have been designed to target various proteases but are limited to extracellular molecules. Only a few have been applied to image intracellular proteases in living cells because most of these probes have limited cell-permeability. Our platform does not need complicated synthetic processes; instead it involves a straightforward peptide synthesis and a simple mixing step with a commercial transfection agent. The transfection agent efficiently delivered the highly quenched fluorogenic probes, comprised of distinctive pairs of dyes and quenchers, to the initiator caspase-8 and the effector caspase-3 in MDA-MB-435 cells, allowing dual-imaging of the activities of both caspases during the apoptotic process induced by TNF-related apoptosis induced ligand (TRAIL). With the combination of multiple fluorogenic probes, this simple platform can be applied to multiplexed imaging of selected intracellular proteases to study apoptotic processes in pathologies or for cell-based high throughput screening systems for drug discovery.
caspase; activatable probe; fluorescence imaging; peptide; transfection agent
Tube and Pelle are essential components in Drosophila Toll signaling pathway. In this study, we characterized a pair of crustacean homologs of Tube and Pelle in Scylla paramamosain, namely, SpTube and SpPelle, and analyzed their immune functions. The full-length cDNA of SpTube had 2052 bp with a 1578 bp open reading frame (ORF) encoding a protein with 525 aa. A death domain (DD) and a kinase domain were predicted in the deduced protein. The full-length cDNA of SpPelle had 3825 bp with a 3420 bp ORF encoding a protein with 1140 aa. The protein contained a DD and a kinase domain. Two conserved repeat motifs previously called Tube repeat motifs present only in insect Tube or Tube-like sequences were found between these two domains. Alignments and structure predictions demonstrated that SpTubeDD and SpPelleDD significantly differed in sequence and 3D structure. Similar to TubeDD, SpTubeDD contained three common conserved residues (R, K, and R) on one surface that may mediate SpMyD88 binding and two common residues (A and A) on the other surface that may contribute to Pelle binding. By contrast, SpPelleDD lacked similar conservative residues. SpTube, insect Tube-like kinases, and human IRAK4 were found to be RD kinases with an RD dipeptide in the kinase domain. SpPelle, Pelle, insect Pelle-like kinases, and human IRAK1 were found to be non-RD kinases lacking an RD dipeptide. Both SpTube and SpPelle were highly expressed in hemocytes, gills, and hepatopancreas. Upon challenge, SpTube and SpPele were significantly increased in hemocytes by Gram-negative or Gram-positive bacteria, whereas only SpPelle was elevated by White Spot Syndrome Virus. The pull-down assay showed that SpTube can bind to both SpMyD88 and SpPelle. These results suggest that SpTube, SpPelle, and SpMyD88 may form a trimeric complex involved in the immunity of mud crabs against both Gram-negative and Gram-positive bacteria.
This paper details an algorithm to simultaneously perform registration of computed tomography (CT) and cone-beam computed (CBCT) images, and image enhancement of CBCT. The algorithm employs a viscous fluid model which naturally incorporates two components: a similarity measure for registration and an intensity correction term for image enhancement. Incorporating an intensity correction term improves the registration results. Furthermore, applying the image enhancement term to CBCT imagery leads to an intensity corrected CBCT with better image quality. To achieve minimal processing time, the algorithm is implemented on a graphic processing unit (GPU) platform. The advantage of the simultaneous optimization strategy is quantitatively validated and discussed using a synthetic example. The effectiveness of the proposed algorithm is then illustrated using six patient datasets, three head-and-neck datasets and three prostate datasets.
Deformable image registration; Multimodal registration; Mutual information; Shading correction; Scatter removal
Enoyl-acyl carrier protein (enoyl-ACP) reductase catalyzes the last step of the elongation cycle in the synthesis of bacterial fatty acids. The Enterococcus faecalis genome contains two genes annotated as enoyl-ACP reductases, a FabI-type enoyl-ACP reductase and a FabK-type enoyl-ACP reductase. We report that expression of either of the two proteins restores growth of an Escherichia coli fabI temperature-sensitive mutant strain under nonpermissive conditions. In vitro assays demonstrated that both proteins support fatty acid synthesis and are active with substrates of all fatty acid chain lengths. Although expression of E. faecalis fabK confers to E. coli high levels of resistance to the antimicrobial triclosan, deletion of fabK from the E. faecalis genome showed that FabK does not play a detectable role in the inherent triclosan resistance of E. faecalis. Indeed, FabK seems to play only a minor role in modulating fatty acid composition. Strains carrying a deletion of fabK grow normally without fatty acid supplementation, whereas fabI deletion mutants make only traces of fatty acids and are unsaturated fatty acid auxotrophs.
The finding that exogenous fatty acids support growth of E. faecalis strains defective in fatty acid synthesis indicates that inhibitors of fatty acid synthesis are ineffective in countering E. faecalis infections because host serum fatty acids support growth of the bacterium.
The internal charge transfer (ICT) type fluoroionophore arylvinyl-bipy (bipy = 2,2′-bipyridyl) is covalently tethered to the spirolactam form of rhodamine to afford fluorescent heteroditopic ligand 4. Compound 4 can be excited in the visible region, the emission of which undergoes sequential bathochromic shifts over an increasing concentration gradient of Zn(ClO4)2 in acetonitrile. Coordination of Zn2+ stabilizes the ICT excited state of the arylvinyl-bipy component of 4, leading to the first emission color shift from blue to green. At sufficiently high concentrations of Zn(ClO4)2, the non-fluorescent spirolactam component of 4 is transformed to the fluorescent rhodamine, which turns the emission color from green to orange via intramolecular fluorescence resonance energy transfer (FRET) from the Zn2+-bound arylvinyl-bipy fluorophore to rhodamine. While this work offers a new design of ratiometric chemosensors, in which sequential analyte-induced emission band shifts result in the sampling of multiple colors at different concentration ranges (i.e. from blue to green to orange as [Zn2+] increases in the current case), it also reveals the nuances of rhodamine spirolactam chemistry that have not been sufficiently addressed in the published literature. These issues include the ability of rhodamine spirolactam as a fluorescence quencher via electron transfer, and the slow kinetics of spirolactam ring-opening effected by Zn2+ coordination.
Zinc; Fluorescent heteroditopic ligand; Rhodamine spirolactam; Fluorescence resonance energy transfer; Photoinduced electron transfer; Internal charge transfer
AIM: To evaluate the efficacy and safety of paclitaxel-nedaplatin combination as a front-line regimen in Chinese patients with metastatic esophageal squamous cell carcinoma (ESCC).
METHODS: A two-center, open-label, single-arm phase II study was designed. Thirty-nine patients were enrolled and included in the intention-to-treat analysis of efficacy and adverse events. Patients received 175 mg/m2 of paclitaxel over a 3 h infusion on 1 d, followed by nedaplatin 80 mg/m2 in a 1 h infusion on 2 d every 3 wk until the documented disease progression, unacceptable toxicity or patient’s refusal.
RESULTS: Of the 36 patients assessable for efficacy, there were 2 patients (5.1%) with complete response and 16 patients (41.0%) with partial response, giving an overall response rate of 46.1%. The median progression-free survival and median overall survival for all patients were 7.1 mo (95%CI: 4.6-9.7) and 12.4 mo (95%CI: 9.5-15.3), respectively. Toxicities were moderate and manageable. Grade 3/4 toxicities included neutropenia (15.4%), nausea (10.3%), anemia (7.7%), thrombocytopenia (5.1%), vomiting (5.1%) and neutropenia fever (2.6%).
CONCLUSION: The combination of paclitaxel and nedaplatin is active and well tolerated as a first-line therapy for patients with metastatic ESCC.
Esophageal squamous cell cancer; Front-line chemotherapy; Paclitaxel; Nedaplatin
A few studies focused on open reduction and internal fixation (ORIF) or nonoperative treatment of displaced 3-part or 4-part proximal humeral fractures in elderly patients have been published, all of whom had a low number of patients. In this meta-analysis of randomized controlled trials (RCTs), we aimed to assess the effect of ORIF or nonoperative treatment of displaced 3-part or 4-part proximal humeral fractures in elderly patients on the clinical outcomes and re-evaluate of the potential benefits of conservative treatment.
We searched PubMed and the Cochrane Central Register of Controlled Trials databases for randomized controlled trials comparing ORIF and nonoperative treatment of displaced 3-part or 4-part proximal humeral fractures in elderly patients. Our outcome measures were the Constant scores.
Results: Three randomized controlled trials with a total of 130 patients were identified and analyzed. The overall results based on fixed-effect model did not support the treatment of open reduction and internal fixation to improve the functional outcome when compared with nonoperative treatment for treating elderly patients with displaced 3-part or 4-part proximal humeral fractures (WMD −0.51, 95% CI: −7.25 to 6.22, P = 0.88, I2 = 0%).
Although our meta-analysis did not support the treatment of open reduction and internal fixation to improve the functional outcome when compared with nonoperative treatment for treating elderly patients with displaced 3-part or 4-part proximal humeral fractures, this result must be considered in the context of variable patient demographics. Only a limited recommendation can be made based on current data. Considering the limitations of included studies, a large, well designed trial that incorporates the evaluation of clinically relevant outcomes in participants with different underlying risks of shoulder function is required to more adequately assess the role for ORIF or nonoperative treatment.
The use of MR contrast agents allows accurate diagnosis by exerting an influence on the longitudinal (T1) or transverse (T2) relaxation time of the surrounding tissue. In this study, we combined the use of iron oxide (IO) particles and nonspecific extracellular gadolinium chelate (Gd) in order to further improve the sensitivity and specificity of lesion detection.
With a 7-Tesla scanner, pre-contrasted, IO-enhanced and dual contrast agent enhanced MRIs were performed in phantom, normal animals, and animal models of lymph node tumor metastases and orthotopic brain tumor. For the dual-contrast (DC) MRI, we focused on the evaluation of T2 weighted DC MRI with IO administered first, then followed by the injection of a bolus of gadolinium diethylenetriamine pentaacetic acid (Gd-DTPA).
Quantified with C/N ratios and MRI relaxometry, the synergistic effect of coordinated administration of Gd-DTPA and IO was observed and confirmed in phantom, normal liver and tumor models. At 30 min after administration of Feridex, Gd-DTPA further decreased T2 relaxation in liver immediately after the injection. Additional administration of Gd-DTPA also immediately increased the signal contrast between tumor and brain parenchyma and maximized the C/N ratio to −4.12 ± 0.71. Dual contrast MRI also enhanced the delineation of tumor borders and small lesions.
DC-MRI will be helpful to improve diagnostic accuracy and decrease the threshold size for lesion detection.
MRI; Gd-DTPA; Iron oxide; RGD; Dual Contrast
Small interfering RNA (siRNA) is an emerging class of therapeutics, working by regulating the expression of a specific gene involved in disease progression. Despite the promises, effective transport of siRNA with minimal side effects remains a challenge. In this study, a non-viral nanoparticle gene carrier has been developed and its efficiency for siRNA delivery and transfection has been validated at both in vitro and in vivo levels. Such a nanocarrier, abbreviated as Alkyl-PEI2k-IO, was constructed with a core of iron oxide (IO) and a shell of alkylated PEI2000 (Alkyl-PEI2k). It was found to be able to bind with siRNA, resulting in well-dispersed nanoparticles with a controlled clustering structure and narrow size distribution. Electrophoresis studies showed that the Alkyl-PEI2k-IOs could retard siRNA completely at N/P ratios above 10, protect siRNA from enzymatic degradation in serum and release complexed siRNA efficiently in the presence of polyanionic heparin. The knockdown efficiency of the siRNA loaded nanocarriers was assessed with 4T1 cells stably expressing luciferase (fluc-4T1) and further, with a fluc-4T1 xenograft model. Significant downregulation of luciferase was observed, and unlike the high molecular weight analogs, the Alkyl-PEI2k coated IOs showed a good biocompatibility. In conclusion, Alkyl-PEI2k-IOs demonstrate highly efficient delivery of siRNA and an innocuous toxic profile, making it a potential carrier for gene therapy.
Biomaterials; Superparamagnetic nanoparticles; Polytehyleneimine; Small interfering RNA
The imaging of sentinel lymph nodes (SLNs), the first defense against primary tumor metastasis, has been considered as an important strategy for noninvasive tracking tumor metastasis in clinics. In this study, we report the development and application of mesoporous silica-based triple-modal nanoprobes that integrate multiple functional moieties to facilitate near-infrared optical, magnetic resonance (MR) and positron emission tomography (PET) imaging. After embedding near-infrared dye ZW800, the nanoprobe was labeled with T1 contrast agent Gd3+ and radionuclide 64Cu through chelating reactions. High stability and long intracellular retention time of the nanoprobes was confirmed by in vitro characterization, which facilitate long-term in vivo imaging. Longitudinal multimodal imaging was subsequently achieved to visualize tumor draining SLNs up to 3 weeks in a 4T1 tumor metastatic model. Obvious differences in uptake rate, amount of particles, and contrast between metastatic and contralateral sentinel lymph nodes were observed. These findings provide very helpful guidance for the design of robust multifunctional nanomaterials in SLNs’ mapping and tumor metastasis diagnosis.
Mesoporous silica nanoparticles; Multimodality imaging; Tumor metastasis; Magnetic resonance imaging; Positron emission tomography; Near-infrared fluorescence imaging
Twenty-one new 4-substituted diarylaniline compounds (DAANs) (Scheme 2, series 13, 14, and 15) were designed, synthesized, and evaluated against wild-type and drug resistant HIV-1 viral strains. As a result, approximately a dozen new DAANs showed high potency with low nano- to sub-nanomolar EC50 values ranging from 0.2 to 10 nM. The three most promising compounds 14e, 14h, and 15h exhibited high potency against wild-type and drug-resistant viral strains with EC50 values at the sub-nanomolar level (0.29–0.87 nM), and were comparable to or more potent than the new NNRTI drug riplivirine (2) in the same assays. Drug-like physicochemical property assessments revealed that the most active DAANs (EC50 <10 nM) have better aqueous solubility (>1–90 μg/mL at pH 7.4 and pH 2) and metabolic stability in vitro than 2, as well as desirable log P values (<5) and polar surface area (PSA) (<140 Å2). These promising results warrant further development of this novel compound class as potential potent anti-AIDS clinical trial candidates.
Betulinic acid derivatives modified at the C28 position are HIV-1entry inhibitors such as compound A43D; however, modified at the C3 position instead of C28 give HIV-1 maturation inhibitor such as bevirimat. Bevirimat exhibited promising pharmacokinetic profiles in clinical trials, but its effectiveness was compromised by the high baseline drug resistance of HIV-1 variants with polymorphism in the putative drug binding site. In an effort to determine whether the viruses with bevirimat resistant polymorphism also altered their sensitivities to the betulinic acid derivatives that inhibit HIV-1 entry, a series of new betulinic acid entry inhibitors were synthesized and tested for their activities against HIV-1 NL4-3 and NL4-3 variants resistant to bevirimat. The results show that the bevirimat resistant viruses were approximately 5- to10-fold more sensitive to three new glutamine ester derivatives (13, 15 and 38) and A43D in an HIV-1 multi-cycle replication assay. In contrast, the wild type NL4-3 and the bevirimat resistant variants were equally sensitive to the HIV-1 RT inhibitor AZT. In addition, these three new compounds markedly improved microsomal stability compared to A43D.
HIV-1; Entry inhibitor; Maturation inhibitor; Betulinic acid; Berivimat; Berivimat-resistance
Background / Objectives
Diet quality indices are increasingly used in nutrition epidemiology as dietary exposures in relation to health outcomes. However, literature on long-term stability of these indices is limited. We aimed to assess the stability of the validated Framingham Nutritional Risk Score (FNRS) and its component nutrients over 8 years as well as the validity of the follow-up FNRS.
Subjects / Methods
Framingham Offspring/Spouse Study women and men (n=1 734) aged 22-76 years wwver 8 years. Individuals' nutrient intake and nutritional risk scores were assessed using 3-day dietary records administered at baseline (1984-1988) and at follow-up (1992-1996). Agreement between baseline and follow-up FNRS and nutrient intakes was evaluated using Bland-Altman method; stability was assessed using intra-class correlation (ICC) and weighted Kappa statistics. The effect of diet quality (as assessed by the FNRS) on cardiometabolic risk factors was evaluated using ANCOVA.
Modest changes from baseline (≤15%) were observed in nutrient intake. Stability coefficients for the FNRS (ICC: women=0.49; men=0.46; P<0.0001) and many nutrients (ICC ≥0.3) were moderate. Over half of women and men (58%) remained in the same or contiguous baseline and follow-up quartile of the FNRS and few (3-4%) shifted >1 quartile. The FNRS was directly associated with BMI in women (P<0.01) and HDL-cholesterol among both women (P<0.001) and men (P<0.01).
The FNRS and its constituent nutrients remained relatively stable over 8 years of follow-up. The stability of diet quality has implications for prospective epidemiological investigations.
long-term stability; dietary quality indices; nutrients
Phase analysis has been developed and validated to measure left-ventricular dyssynchrony from gated SPECT myocardial perfusion imaging. The purpose of this study is to evaluate its performance in regions with perfusion defects.
A special version of the eXtended CArdiac Torso digital phantom was developed to track B-spline points in each temporal frame. A region of 35 B-spline points in the inferior wall with normal and abnormal perfusion uptakes were simulated. Phase shifts were simulated in the same region, representing dyssynchronous contraction. Gated SPECT data were analyzed using a modified phase analysis algorithm, which tracked the same 35 B-spline points to calculate their phases.
Phases and phase shifts measured in the B-spline points with perfusion uptake in the range of 50%–10% did not significantly differ from those measured in the same B-spline points with normal perfusion uptake.
Phase analysis can accurately measure phases in regions with abnormal perfusion uptake as low as 10% of the perfusion uptake in the normal regions, which corresponded to a regional signal-to-noise ratio (SNR) of 12.0 or greater. In 42 consecutive patients with myocardial infarction >20% of the left ventricle, only two patients had a SNR within the perfusion defects below that threshold.
Myocardial perfusion imaging; gated SPECT; phase analysis; LV dyssynchrony
Despite their immense potential in biomedicine, carbon nanomaterials suffer from inefficient dispersion and biological activity in vivo. Here we utilize a single, yet multifunctional, hyaluronic acid-based biosurfactant to simultaneously disperse nanocarbons and target single-walled carbon nanotubes (SWCNTs) to CD44 receptor positive tumor cells with prompt uptake. Cellular uptake was monitored by intracellular enzyme-activated fluorescence and localization of SWCNTs within cells was further confirmed by Raman mapping. In vivo photoacoustic, fluorescence and positron emission tomography imaging of coated SWCNTs display high tumor targeting capability while providing long-term, fluorescence molecular imaging of targeted enzyme events. By utilizing a single biomaterial surfactant for SWCNT dispersion without additional bioconjugation, we designed a facile technique that brings nanocarbons closer to their biomedical potential.
Carbon nanomaterials; one-step functionalization; hyaluronic acid; nanotubes; molecular
Obesity affects one in three American adult women and is associated with overall mortality and major morbidities. A composite diet index to evaluate total diet quality may better assess the complex relationship between diet and obesity, providing insights for nutrition interventions. The purpose of the present investigation was to determine whether diet quality, defined according to the previously validated Framingham nutritional risk score (FNRS), was associated with the development of overweight or obesity in women. Over 16 years, we followed 590 normal-weight women (BMI < 25 kg/m2), aged 25 to 71 years, of the Framingham Offspring and Spouse Study who presented without CVD, cancer or diabetes at baseline. The nineteen-nutrient FNRS derived from mean ranks of nutrient intakes from 3d dietary records was used to assess nutritional risk. The outcome was development of overweight or obesity (BMI ≥ 25 kg/m2) during follow-up. In a stepwise multiple logistic regression model adjusted for age, physical activity and smoking status, the FNRS was directly related to overweight or obesity (P for trend= 0·009). Women with lower diet quality (i.e. higher nutritional risk scores) were significantly more likely to become overweight or obese (OR 1·76; 95% CI 1·16, 2·69) compared with those with higher diet quality. Diet quality, assessed using a comprehensive composite nutritional risk score, predicted development of overweight or obesity. This finding suggests that overall diet quality be considered a key component in planning and implementing programmes for obesity risk reduction and treatment recommendations.
Diet quality; Nutritional risk score; Obesity; BMI; Dietary quality index
Noninvasive imaging techniques have been considered important strategies in the clinic to monitor tumor early response to therapy. In the present study, we applied RGD peptides conjugated to iron oxide nanoparticles (IONP-RGD) as contrast agents in magnetic resonance imaging (MRI) to noninvasively monitor the response of a vascular disrupting agent VEGF121/rGel in an orthotopic glioblastoma model. RGD peptides were firstly coupled to IONPs coated with a crosslinked PEGylated amphiphilic triblock copolymer. In vitro binding assays confirmed that cellular uptake of particles was mainly dependent on the interaction between RGD and integrin αvβ3 of human umbilical vein endothelial cells (HUVEC). The tumor targeting of IONP-RGD was observed in an orthotopic U87 glioblastoma model. Finally, noninvasive monitoring of the tumor response to VEGF121/rGel therapy at early stages of treatment was successfully accomplished using IONP-RGD as a contrast agent for MRI, a superior method over common anatomical approaches which are based on tumor size measurements. This preclinical study can accelerate anticancer drug development and promote clinical translation of nanoprobes.
Magnetic resonance imaging (MRI); Iron oxide nanoparticles (IONPs); RGD peptides; Tumor targeting; Therapy response
While idiopathic pulmonary fibrosis (PF) is a devastating lung disease, the management of PF including effective monitoring of disease progression remains a challenge. Herein, we introduce a novel, fast and ultra-sensitive metalloproteinase (MMP) activatable optical probe, named MMP-P12, to non-invasively monitor PF progression and response to PF treatment. A bleomycin (BLM)-induced mouse PF model was subjected non-invasively to optical imaging at various time points after BLM treatment. Mouse PF model developed fibrosis during 21 days of experimental period, and the progression of PF was well correlated with the step-wise increase of MMP-2 expression as examined by quantitative RT-PCR and western blot analysis on the 7-, 14-and 21-day post-BLM administration. On these days, MMP-activated fluorescence images were acquired in vivo and ex vivo. Signal quantification showed time-dependent lung-specific incremental increases in fluorescence signals. As a treatment for PF, secretoglobin 3A2 was daily administered intravenously for five days starting day seven of BLM administration, which resulted in reduced MMP-2 activity and reduction of PF as previously demonstrated. Importantly, the fluorescence signal that reflected MMP activity also decreased in intensity. In conclusion, MMPs may play an important role in PF development and MMP-P12 probe could be a promising tool for PF detection, even at an early stage of the disease as well as an indicator of therapy response.
Pulmonary fibrosis; Matrix metalloproteinase; Optical imaging; Activatable probe; Secretoglobin 3A2
Human empathy is not merely a resonance with others’ physical condition, but is modulated by social factors. Using functional magnetic resonance imaging, the present study demonstrated an increased brain empathic response to others in pain when they received no rather than a large reward, with increments of the ACC, aMCC, insula and postcentral gyrus in the pain matrix and temporoparietal junction. Thus, pain target’s financial situation modulated brain empathic responses in the pain matrix based on an understanding of the situation pain target faces.
empathy; pain; monetary reward; aMCC; insula
This paper presents a novel approach that three-dimensionally visualizes and evaluates stenoses in human coronary arteries by using harmonic skeletons. A harmonic skeleton is the center line of a multi-branched tubular surface extracted based on a harmonic function, which is the solution of the Laplace equation. This skeletonization method guarantees smoothness and connectivity and provides a fast and straightforward way to calculate local cross-sectional areas of the arteries, and thus provides the possibility to localize and evaluate coronary artery stenosis, which is a commonly seen pathology in coronary artery disease.
Image morphing, or image interpolation in the time domain, deals with the metamorphosis of one image into another. In this paper, a new class of image morphing algorithms is proposed based on the theory of optimal mass transport. The L2 mass moving energy functional is modified by adding an intensity penalizing term, in order to reduce the undesired double exposure effect. It is an intensity-based approach and, thus, is parameter free. The optimal warping function is computed using an iterative gradient descent approach. This proposed morphing method is also extended to doubly connected domains using a harmonic parameterization technique, along with finite-element methods.
Image interpolation; image morphing; image warping; mass preserving mapping; Monge–Kantorovich flow; optimal transport
This paper presents a new algorithm for non-rigid registration between two doubly-connected regions. Our algorithm is based on harmonic analysis and the theory of optimal mass transport. It assumes an underlining continuum model, in which the total amount of mass is exactly preserved during the transformation of tissues. We use a finite element approach to numerically implement the algorithm.
Hepatocellular carcinoma (HCC) is one of the most fatal cancers. In almost all populations, males have a higher HCC rate than females. Here we sought to explore the roles and mechanisms of acetylcholine (Ach) and androgen receptor (AR) on regulating the fate determinations of HCC. Ach activated AR and promoted its expression in HCC cells. Ach enhanced HCC cell migration and invasion but inhibited their apoptosis. Ach had no obvious effects on the migration, invasion, or apoptosis in AR-negative HCC cells. Elevation of migration and invasion induced by Ach was eliminated in AR-knockdown HCC cells. In contrast, Ach stimulated the migration and invasion but suppressed apoptosis in AR over-expressed HCC cells. Additionally, AR agonist R1881 promoted the migration and invasion but reduced the apoptosis of SNU-449 cells, whereas AR antagonist casodex inhibited the migration and invasion but stimulated the apoptosis of SNU-449 cells. STAT3 and AKT phosphorylation was activated by Ach in HCC cells. Collectively, these data suggest that Ach activates STAT3 and AKT pathways and acts on AR to promote the migration and invasion but inhibit the apoptosis of HCC cells. This study thus provides novel insights into carcinogenesis of liver cancer by local interaction between neurotransmitter Ach and hormone receptor AR in HCC.
ferritin; fluorescent probes; imaging agents; metalloenzymes; nanoparticles
caspases; fluorescence; imaging agents; nanoparticles; proteolysis