Cisplatin is one of the most commonly used chemotherapy agent for lung cancer. The therapeutic efficacy of cisplatin is limited by the development of resistance.
In this study, we test the effect of RNA interference (RNAi) targeting Fanconi anemia (FA)/BRCA pathway upstream genes on the sensitivity of cisplatin-sensitive (A549 and SK-MES-1) and -resistant (A549/DDP) lung cancer cells to cisplatin.
Using small interfering RNA (siRNA), knockdown of FANCF, FANCL, or FANCD2 inhibited function of the FA/BRCA pathway in A549, A549/DDP and SK-MES-1 cells, and potentiated sensitivity of the three cells to cisplatin. The extent of proliferation inhibition induced by cisplatin after knockdown of FANCF and/or FANCL in A549/DDP cells was significantly greater than in A549 and SK-MES-1 cells, suggesting that depletion of FANCF and/or FANCL can reverse resistance of cisplatin-resistant lung cancer cells to cisplatin. Furthermore, knockdown of FANCL resulted in higher cisplatin sensitivity and dramatically elevated apoptosis rates compared with knockdown of FANCF in A549/DDP cells, indicating that FANCL play an important role in the repair of cisplatin-induced DNA damage.
Knockdown of FANCF, FANCL, or FANCD2 by RNAi could synergize the effect of cisplatin on suppressing cell proliferation in cisplatin-resistant lung cancer cells through inhibition of FA/BRCA pathway.
Chemoresistance; Cisplatin; Lung cancer; RNA interference; Fanconi anemia/BRCA pathway; Core complex
Accumulation of lipofuscin in the
retina is associated with pathogenesis
of atrophic age-related macular degeneration and Stargardt disease.
Lipofuscin bisretinoids (exemplified by N-retinylidene-N-retinylethanolamine) seem to mediate lipofuscin toxicity.
Synthesis of lipofuscin bisretinoids depends on the influx of retinol
from serum to the retina. Compounds antagonizing the retinol-dependent
interaction of retinol-binding protein 4 (RBP4) with transthyretin
in the serum would reduce serum RBP4 and retinol and inhibit bisretinoid
formation. We recently showed that A1120 (3), a potent
carboxylic acid based RBP4 antagonist, can significantly reduce lipofuscin
bisretinoid formation in the retinas of Abca4–/– mice. As part of the NIH
Blueprint Neurotherapeutics Network project we undertook the in vitro exploration to identify novel conformationally flexible and constrained
RBP4 antagonists with improved potency and metabolic stability. We
also demonstrate that upon acute and chronic dosing in rats, 43, a potent cyclopentyl fused pyrrolidine antagonist, reduced
circulating plasma RBP4 protein levels by approximately 60%.
Purpose: This study aimed to investigate the synergistic anti-tumor effects of tumstatin 185-191 and cisplatin in non-small cell lung carcinoma cells (NSCLC) (A549 cells and cisplatin resistant A549/DDP cells), and the potential role of Akt signaling pathway was also explored. Methods: A549 or A549/DDP cells were treated with Tum185-191 or Tum185-191 plus cisplatin. Cell viability was assessed by modified MTT assay. 50% inhibiting concentration (IC50) and reversing drug-resistance index (RI) of chemotherapeutics were determined by MTT assay. Cell apoptosis was measured by Hoechst 33258 staining and flow cytometry. The activation of Akt signaling pathway was evaluated by immunocytochemistry and Western blot assay. Results: Tum185-191 inhibited the proliferation of A549 cells and A549/DDP cells. In the presence of Tum185-191 (20 and 40 μM), IC50 of cisplatin reduced significantly in A549 cells and A549/DDP cells. Combined use of tumstatin 185-191 and cisplatin exerted synergistic effects in promoting apoptosis. A549 and A549/DDP cells had a high expression of p-Akt, and Tum185-191, but not cisplatin, significantly inhibited p-Akt expression. Combined use of cisplatin and Tum185-191 failed to further inhibit p-Akt expression. After Tum185-191 treatment, the increased p-Akt expression was observed at 15 min, peaked at 30-60 min, but disappeared at 120 min. Conclusion: Tum185-191 increases the apoptosis, inhibit the proliferation, enhance the sensitivity of A549 cells to cisplatin and also partly reverse the resistance of A549-DDP cells to cisplatin, which is at least partially mediated by inactivating Akt pathway. These findings provide evidence for the chemotherapy of NSCLC with Tum185-191 and cisplatin.
Non-small cell lung carcinoma cells; Tumstain 185-191; cisplatin; Akt
Antibody-dependent pathogenicity is suggested in multiple sclerosis (MS) by intrathecal immunoglobulin production, IgG and complement deposition in the most common immunopathological lesion subtype (pattern II), and by a recent report that 47% of MS patients’ sera contain a glial potassium-channel-specific-IgG(inwardly-rectifying, Kir4.1). Our study’s aims were to determine, in MS serum and CSF, the frequency and specificity of Kir4.1-binding-IgG and, in demyelinating MS lesions, whether Kir4.1-immunoreactivity is retained or lost.
We tested by ELISA(Kir4.1-peptide 83–120) sera from 286 clinically and geographically diverse MS patients (229 population-based and 57 clinic-based),99 healthy controls and 109 disease controls, and cerebrospinal fluid [CSF] from 25 MS and 22 controls. CSFs and clinic-based MS-subset serum (50)were tested on functional Kir4.1-expressing cells, using methodologies validated for detecting clinically-pertinent neural plasma membrane-reactive autoantibodies: immunofluorescence and immunoprecipitation (solubilized recombinant human Kir4.1). We evaluated Kir4.1-immunoreactivity in brain from 15 archival histopathologically-confirmed MS cases(22 plaques: 8 early active, 8 inactive, 6 remyelinated; 13 periplaque regions)and compared 3 non-neurological cases (8 normal-appearing white/gray matter regions).
Kir4.1-peptide-ELISA reactivity was rare and did not differ significantly for 286 MS or 208 control sera (both 1%); no CSF was positive. IgGin 0/50 clinic-based MS sera immunoprecipitated Kir4.1, but control Kir4.1-specific-IgG did. By immunofluorescence,1/50 MS sera yielded faint plasmalemmal staining on both Kir4.1-expressing and non-expressing cells; 16/50 bound faintly to intracellular components. In all cases, IgG binding was quenched by absorption with liver powder or non-transfected cell lysates. Control Kir4.1-specific-IgG binding was quenched only by Kir4.1 protein-containing lysates. IgG in 0/25 MS CSFs bound to Kir4.1-transfected cells, live or fixed. Glial Kir4.1-immunoreactivity was increased relative to baseline normal brain expression (3 controls) in early active and remyelinated MS lesions, and in periplaque white matter (15 patients).
We did not find Kir4.1-specific-IgG in MS sera or CSF, nor Kir4.1 loss from glial cells in active demyelinating MS lesions. Serological testing for Kir4.1-IgG is unlikely to aid MS diagnosis. The target antigen of MS remains elusive.
The National Institutes of Health, the National Multiple Sclerosis Society and the Mayo Clinic Robert and Arlene Kogod Center on Aging.
Studies show a strong association between dementia and lower urinary tract symptoms (LUTS). The aim of this study was to investigate whether LUTS are a risk factor for cognitive impairment. We enrolled 50-year-old and older subjects with LUTS (LUTS[+]) (n = 6801) and controls without LUTS (LUTS[−]) (n = 20,403) from Taiwan's National Health Insurance Research Database. LUTS, dementia, and other confounding factors are defined by International Classification of Diseases, Ninth Revision, Clinical Modification Codes. Participants were recruited from 2000 to 2004 and then followed up until death or the end of 2011. The outcome was the onset of dementia, which was assessed using Poisson regression analysis, Cox hazards models, and Kaplan-Meier survival curves. The incidence of dementia was significantly higher in the LUTS[+] group than in the LUTS[−] group (124.76 versus 77.59/1000 person-years). The increased risk of dementia related to LUTS remained significant after adjustment for potential confounders (adjusted hazard ratio (AHR): 1.61, 95% confidence interval (CI) 1.47–1.76, P < 0.0001) and higher than that related to cerebrovascular disease (AHR: 1.43, 95% CI 1.26–1.61, P < 0.0001). The outcome suggests the need for early screening and appropriate intervention to help prevent cognitive impairment of patients with LUTS.
Medical imaging is becoming a vital component of war on cancer. Tremendous amounts of medical image data are captured and recorded in a digital format during cancer care and cancer research. Facing such an unprecedented volume of image data with heterogeneous image modalities, it is necessary to develop effective and efficient content-based medical image retrieval systems for cancer clinical practice and research. While substantial progress has been made in different areas of content-based image retrieval (CBIR) research, direct applications of existing CBIR techniques to the medical images produced unsatisfactory results, because of the unique characteristics of medical images. In this paper, we develop a new multimodal medical image retrieval approach based on the recent advances in the statistical graphic model and deep learning. Specifically, we first investigate a new extended probabilistic Latent Semantic Analysis model to integrate the visual and textual information from medical images to bridge the semantic gap. We then develop a new deep Boltzmann machine-based multimodal learning model to learn the joint density model from multimodal information in order to derive the missing modality. Experimental results with large volume of real-world medical images have shown that our new approach is a promising solution for the next-generation medical imaging indexing and retrieval system.
content-based image retrieval; multi-modal and content-based medical image retrieval; extended probabilistic latent semantic analysis; deep learning; deep boltzmann machine
Despite the large number of published papers analyzing the prognostic role of Ki-67 in NSCLC, it is still not considered an established factor for routine use in clinical practice. The present meta-analysis summarizes and analyses the associations between Ki-67 expression and clinical outcome in NSCLC patients.
PubMed, Cochrane, and Embase databases were searched systematically using identical search strategies. The impacts of Ki-67 expression on survival in patients with NSCLC and NSCLC subtypes were evaluated. Furthermore, the association between Ki-67 expression and the clinicopathological features of NSCLC were evaluated.
In total, 32 studies from 30 articles met the inclusion criteria, involving 5600 patients. Meta-analysis results suggested that high Ki-67 expression was negatively associated with overall survival (OS; HR = 1.59, 95 % CI 1.35-1.88, P < 0.001) and disease-free survival (DFS; HR = 2.21, 95 % CI 1.43-3.42, P < 0.001) in NSCLC patients. Analysis of the different subgroups of NSCLC suggested that the negative association between high Ki-67 expression and OS and DFS in Asian NSCLC patients was stronger than that in non-Asian NSCLC patients, particularly in early-stage (Stage I-II) adenocarcinoma (ADC) patients. Additionally, while high expression was more common in males, smokers, and those with poorer differentiation, there was no correlation between high Ki-67 expression and age or lymph node status. Importantly, significant correlations between high Ki-67 expression and clinicopathological features (males, higher tumor stage, poor differentiation) were seen only in Asian NSCLC patients.
The present meta-analysis indicated that elevated Ki-67 expression was associated with a poorer outcome in NSCLC patients, particularly in early-stage Asian ADC patients. Studies with larger numbers of patients are needed to validate our findings.
Electronic supplementary material
The online version of this article (doi:10.1186/s12885-015-1524-2) contains supplementary material, which is available to authorized users.
Ki-67; Meta-analysis; Non-small cell lung cancer; Prognostic value
Histone H2B monoubiquitination is a key histone modification that has significant effects on chromatin higher-order structure and gene transcription. Multiple biological processes have been suggested to be tightly related to the dynamics of H2B monoubiquitination. However, a comprehensive understanding of biological roles of H2B monoubiquitination is still poorly understood. In the present study, we developed an efficient tool to disrupt endogenous H2B monoubiquitination levels by using an H2BK120R mutant construct expressed in human cells. Genome-wide microarray analysis of these cells revealed a potential global view of biological functions of H2B monoubiquitination. Bioinformatics analysis of our data demonstrated that while H2B monoubiquitination expectedly affected a number of previously reported biological pathways, we also uncovered the influence of this histone modification on many novel biological processes. Therefore, our work provided valuable information for understanding the role of H2B monoubiquitination and indicated potential directions for its further studies.
Accumulating evidence suggests that long non-coding RNA (lncRNA) HOTAIR participates in many types of cancer such as gastric cancer and may confer malignant phenotype to tumor cells. Fluorouracil and platinum combination chemotherapy is the first line therapy for gastric cancer. However, it is still unknown whether HOTAIR influences the outcome of cancer patients treated with chemotherapy. This study aimed to evaluate the association of HOTAIR expression with the prognosis of patients with advanced gastric adenocarcinoma (GA) receiving fluorouracil and platinum based chemotherapy. We examined the levels of HOTAIR in 168 GA samples using quantitative real-time PCR and analyzed its relationship with clinical features and prognosis of patients with advanced GA treated with fluorouracil and platinum based chemotherapy. Compared with paracancerous tissues, HOTAIR was significantly upregulated in GA tissues, especially in more advanced cases. High HOTAIR expression was an independent poor prognostic factor for patients with advanced GA. Further stratification analyses revealed that the association between HOTAIR expression and survival in patients with advanced GA remained significant in the subgroup of patients with TNM stages IIIA and IIIB, poorly differentiated, and smaller tumors. In conclusion, our results provide first evidence that HOTAIR may be served as a biomarker that predicts which patient with advanced GA will benefit from fluorouracil and platinum combination chemotherapy.
Gastric cancer; long noncoding RNA; HOTAIR; chemotherapy; prognosis
Numerous studies focusing on genetic variants in order to find cetuximab subpopulation biomarkers have emerged, yet the significance of each biomarker is diverse. Based on these results, we carried out a meta-analysis to assess the correlation between epidermal growth factor (EGF) A61G polymorphism and clinical outcomes of metastatic colorectal cancer (mCRC) patients treated with cetuximab. We aim to prove that EGF polymorphisms may be potential biomarkers for cetuximab therapeutic strategies. We identified 6 previously published studies including 569 patients treated with cetuximab-based regimens. Outcomes included clinical response, progression-free survival (PFS), and overall survival (OS). GG homozygote showed association with better response rates (GG vs. AA+AG, OR = 2.82; 95% CI = 1.58-5.04) and than AA+AG genotypes. This meta-analysis showed that mCRC patients harboring GG genotype of EGF A61G polymorphism inclined to have a better response rate with cetuximab treatment.
EGF; polymorphism; mCRC
The acute torsion of wandering spleen is a very rare disease characterized by acute abdominal pain. Without early surgical intervention, wandering spleen can lead to splenic infarction or rupture. However, early clinical diagnosis is very difficult, so imaging modalities play an important role. We present a case of acute abdominal pain due to torsion of the wandering spleen in a 17-year-old girl, diagnosed by computed tomography and effectively managed by splenectomy for splenic infarction.
Wandering spleen; splenectomy; splenic infarction
Neck dissection is the most definitive and effective treatment for head and neck cancer. This systematic review aims to compare the efficacy and surgical outcomes of neck dissection between the harmonic scalpel and conventional surgical techniques and conduct a quantitative meta-analysis of the randomized trials.
Randomized controlled trials (RCTs) were identified from the major electronic databases (MEDLINE, EMBASE and Cochrane Library) using the keywords ‘‘harmonic scalpel’’ and ‘‘neck dissection,’’ and a quantitative meta-analysis was conducted. The operative time and intraoperative bleeding were the primary outcome measures, and other parameters assessed included the drainage fluid volume and length of hospital stay.
Seven trials that met the inclusion criteria included 406 neck dissection cases (201 in the harmonic scalpel group). Compared with conventional surgical techniques, the HS group had an operative time that was significantly reduced by 29.3 minutes [mean difference: -29.29; 95% CI = (-44.26, -14.32); P=0.0001], a reduction in intraoperative bleeding by 141.1 milliliters [mean difference: -141.13; 95% CI = (-314.99, 32.73); P=0.11], and a reduction in drainage fluid volume by 64.9 milliliters [mean difference: -64.86; 95% CI = (-110.40, -19.32); P=0.005] , but it is not significant after removal of studies driving heterogeneity. There was no significant difference in the length of the hospital stay [mean difference: -0.21; 95% CI = (-0.48, 0.07); P=0.14].
This systematic review showed that using the harmonic scalpel for neck dissection significantly reduces the operative time and drainage fluid volume and that it is not associated with an increased length of hospital stay or perioperative complications. Therefore, the harmonic scalpel method is safe and effective for neck dissection. However, the statistical heterogeneity was high. Further studies are required to substantiate our findings.
An excitatory peptide, di16a, with 49 amino acids and ten cysteine residues was purified and characterized from the venom of Conus distans. Five AA residues were modified: one γ–carboxyglutamate (Gla), and four hydroxyproline (Hyp) residues. A cDNA clone encoding the precursor for the peptide was characterized; the peptide has a novel cysteine framework and a distinctive signal sequence that differs from any other conotoxin superfamily. The peptide was chemically synthesized and folded, and synthetic and native materials were shown to co-elute. Injection of the synthetic peptide causes a hyperexcitable phenotype in mice greater than three weeks of age at lower doses, and lethargy at higher doses. The peptide defines both a previously-uncharacterized gene superfamily of conopeptides, and a new Cys pattern with three vicinal Cys residues.
Conus venom; Conus peptide; Conotoxin; γ–Carboxyglutamate; Hydroxyproline; Conus distans
Efficient DNA double-strand break (DSB) repair is critical for the maintenance of genome stability. Unrepaired or misrepaired DSBs cause chromosomal rearrangements that can result in severe consequences, such as tumorigenesis. RAD6 is an E2 ubiquitin-conjugating enzyme that plays a pivotal role in repairing UV-induced DNA damage. Here, we present evidence that RAD6 is also required for DNA DSB repair via homologous recombination (HR) by specifically regulating the degradation of heterochromatin protein 1α (HP1α). Our study indicates that RAD6 physically interacts with HP1α and ubiquitinates HP1α at residue K154, thereby promoting HP1α degradation through the autophagy pathway and eventually leading to an open chromatin structure that facilitates efficient HR DSB repair. Furthermore, bioinformatics studies have indicated that the expression of RAD6 and HP1α exhibits an inverse relationship and correlates with the survival rate of patients.
Myeloid dendritic cells (mDCs) play an important role in autoimmune diseases. However, the role of blood CD1c+ myeloid dendritic cells 1 (mDC1s), the subset of human blood mDCs, is not well understood in noninfectious uveitis.
Fresh peripheral blood samples from human noninfectious uveitis patients (n = 32) and healthy controls (HCs) (n = 64) were stained with FITC-Lineage 1 (Lin1), PERCP-HLADR, and PE-CD1c antibodies. The levels of mDC1 were quantified by using flow cytometric analysis. Longitudinal data from patients (n = 16) were analyzed to correlate the levels of mDC1 with disease activity.
Blood CD1c+ mDC1 and its subpopulation, CD1chi mDC1, were increased in uveitis patients compared with HCs. Longitudinal data demonstrated that both the CD1c+ mDC1 and CD1chi mDC1 subpopulation reflected a dynamic change in clinical uveitis activity: CD1c expression was increased in active uveitis but decreased when uveitis became inactive.
Given these observations, an alteration in blood CD1c+ mDC1 and the CD1chi mDC1 subpopulation could be a potential biomarker to monitor clinical uveitis activity within patients.
Our study has found that the CD1c+ mDC1 and CD1chi mDC1 subpopulation sensitively reflect the longitudinal disease activity in uveitis. This finding suggests these populations could be used to monitor disease activity and as a marker of treatment efficacy in a clinical setting.
CD1c+ mDC1; CD1chi mDC1 subpopulation; noninfectious uveitis; autoimmune disease
Human eukaryotic prohibitin (prohibitin-1 and prohibitin-2) is a membrane protein with different cellular localizations. It is involved in multiple cellular functions, including energy metabolism, proliferation, apoptosis, and senescence. The subcellular localization of prohibitin may determine its functions. Membrane prohibitin regulate the cellular signaling of membrane transport, nuclear prohibitin control transcription activation and the cell cycle, and mitochondrial prohibitin complex stabilize the mitochondrial genome and modulate mitochondrial dynamics, mitochondrial morphology, mitochondrial biogenesis, and the mitochondrial intrinsic apoptotic pathway. Moreover, prohibitin can translocates into the nucleus or the mitochondria under apoptotic signals and the subcellular shuttling of prohibitin is necessary for apoptosis process. Apoptosis is the process of programmed cell death that is important for the maintenance of normal physiological functions. Consequently, any alteration in the content, post-transcriptional modification (i.e. phosphorylation) or the nuclear or mitochondrial translocation of prohibitin may influence cell fate. Understanding the mechanisms of the expression and regulation of prohibitin may be useful for future research. This review provides an overview of the multifaceted and essential roles played by prohibitin in the regulation of cell survival and apoptosis.
Prohibitin; Survival; Apoptosis
accumulation of pathologic protein fragments is common in neurodegenerative
disorders. We have recently identified in Alzheimer’s disease
(AD) the aggregation of the U1-70K splicing factor and abnormal RNA
processing. Here, we present that U1-70K can be cleaved into an N-terminal
truncation (N40K) in ∼50% of AD cases, and the N40K abundance
is inversely proportional to the total level of U1-70K. To map the
cleavage site, we compared tryptic peptides of N40K and stable isotope
labeled U1-70K by liquid chromatography–tandem mass spectrometry
(MS), revealing that the proteolysis site is located in a highly repetitive
and hydrophilic domain of U1-70K. We then adapted Western blotting
to map the cleavage site in two steps: (i) mass spectrometric analysis
revealing that U1-70K and N40K share the same N-termini and contain
no major modifications; (ii) matching N40K with a series of six recombinant
U1-70K truncations to define the cleavage site within a small region
(Arg300 ± 6 residues). Finally, N40K expression led to substantial
degeneration of rat primary hippocampal neurons. In summary, we combined
multiple approaches to identify the U1-70K proteolytic site and found
that the N40K fragment might contribute to neuronal toxicity in Alzheimer’s
mass spectrometry; LC−MS/MS; proteomics; neurodegenerative disease; Alzheimer’s
disease; stable isotope labeling; proteolytic cleavage; U1 snRNP; U1-70K; RNA splicing
In order to provide Chinese clinicians with guidelines for the management of massive blood loss, we investigated the correlation between the frequency of blood tests and the mortality rate in patients undergoing massive blood transfusion (MBT). The aim of this study is to provide Chinese clinicians with guidelines for the management of massive blood loss. We retrospectively reviewed the medical records of patients who underwent massive blood transfusion (MBT) from 20 tertiary hospitals in 5 regions of China. The frequency of blood tests performed within 24 or 72 hours was compared between patients infused with < 10 and ≥ 10 U of red blood cells (RBC). The correlation between the frequency of blood tests and the mortality rate was determined. A high frequency of blood tests was associated with a low mortality rate in MBT cases. The frequency of all blood tests performed within 24 hours was negatively correlated with the mortality rate in patients infused with ≥ 10 U of RBC, while the frequency of blood coagulation tests performed within 72 hours was negatively correlated with the mortality rate in both patients infused with ≥ 10 and < 10 U of RBC. In conclusions: Measuring the blood indices frequently within the first 24 hours of MBT links to lower mortality rate. Coagulation indices in MBT patients should be closely monitored in the long term to help improve survival.
Frequency of blood tests; massive blood transfusion; mortality rate; retrospective analysis; blood coagulation; massive transfusion protocols; transfusion; coagulopathy; acidosis; hypothermia
Adenoviral vector is an efficient tool for gene transfer. Protein expression is regulated by a number of factors, but the regulation by gene copy number remains to be investigated further.
Assessed by flow cytometry, we demonstrated a significant linear correlation between average fluorescence intensity of green fluorescent protein (GFP) and a wide range of multiplicity of infection (MOI), spanning from 0.01 to 200. Average GFP intensity was calculated by mean fluorescence intensity (MFI) × percentage of infection (POI) (MFI × POI) and the correlation was observed in cells transduced with GFP-expressing adenoviral vector driven either by a cytomegalovirus (CMV) promoter for 3 to 6 h or by a human phosphoglycerate kinase (PGK) promoter for 18 to 24 h. Factors impacting this linear correlation include MOI of viral vector, strength of promoter driving GFP expression, cell type transduced and incubation time after gene transfer. We also found that weak GFP signals could be interfered by background signals, whereas strong GFP signals could overshot the detection limitation of the flow cytometer and resulted in a deviation from linearity which was prevented by adjusting the setting in flow cytometer. Moreover, we compared promoter strength as measured by MFI × POI and found that the relative activity of CMV promoter to PGK promoter was 20 to 47 folds in A549 cells and 32 to > 100 folds in H1299 cells.
The linear correlation between MFI × POI and a wide range of adenoviral MOI provides an efficient method to investigate factors regulating protein expression and to estimate virus titers.
Electronic supplementary material
The online version of this article (doi:10.1186/s12929-015-0137-z) contains supplementary material, which is available to authorized users.
Adenovirus; Flow cytometry; Average fluorescence intensity; Linear correlation; Promoter activity
Hedgehog (Hh) signaling plays vital roles in animal development and tissue homeostasis, and its misregulation causes congenital diseases and several types of cancer. Suppressor of Fused (Su(fu)) is a conserved inhibitory component of the Hh signaling pathway, but how it is regulated remains poorly understood. Here we demonstrate that in Drosophila Hh signaling promotes downregulation of Su(fu) through its target protein HIB (Hh-induced BTB protein). Interestingly, although HIB-mediated downregulation of Su(fu) depends on the E3 ubiquitin ligase Cul3, HIB does not directly regulate Su(fu) protein stability. Through an RNAi-based candidate gene screen, we identify the spliceosome factor Crooked neck (Crn) as a regulator of Su(fu) level. Epistasis analysis indicates that HIB downregulates Su(fu) through Crn. Furthermore, we provide evidence that HIB retains Crn in the nucleus, leading to reduced Su(fu) protein level. Finally, we show that SPOP, the mammalian homologue of HIB, can substitute HIB to downregulate Su(fu) level in Drosophila. Our study suggests that Hh regulates both Ci and Su(fu) levels through its target HIB, thus uncovering a novel feedback mechanism that regulates Hh signal transduction. The dual function of HIB may provide a buffering mechanism to fine-tune Hh pathway activity.
Hh; Su(fu); HIB; Crn; SPOP; Cul3
Previous research of Bornean Micronectidae Jaczewski, 1924 (pygmy water boatmen) is summarized based on the data from the literature and recent work. All the Bornean micronectids belong to the genus Micronecta Kirkaldy, 1897. Descriptions or redescriptions and a key to the eight species, which have so far been found in Borneo are presented, namely Micronecta
decorata Lundblad, 1933, Micronecta
ludibunda Breddin, 1905, Micronecta
sp. n., Micronecta
sp. n., Micronecta
lumutensis Chen, Nieser & Lansbury, 2008, Micronecta
skutalis Nieser & Chen, 1999, Micronecta
kymatista Nieser & Chen, 1999) and Micronecta
quadristrigata Breddin, 1905. The synonyms are indicated under each species. To facilitate identification, illustrations and habitus photos are provided. The faunistic components of Micronectidae in Borneo are discussed from a zoogeographic point of view.
Hemiptera; Micronectidae; Micronecta; new species; key; Borneo
Gastric cancer (GC) is the second most common cause of cancer-related death with limited serum biomarkers for diagnosis and prognosis. Netrin-4 (Ntn4) is a laminin-related secreted molecule found to regulate tumor progression and metastasis. However, it is completely unknown whether Ntn4 has roles in GC development. Here, we first reported Ntn4 knockdown significantly suppressed cell proliferation and motility, while overexpression or addition of exogenous Ntn4 reversed these effects. In addition, Ntn4 receptor, neogenin (Neo) was also found highly expressed in GC cells and mediated the Ntn4-induced cell proliferation and invasion. Moreover, Ntn4 or Neo silencing decreased the phosphorylation of Stat3, ERK, Akt and p38, indicating multi-oncogenic pathways (Jak/Stat, PI3K/Akt, and ERK/MAPK) were involved in Ntn4-induced effects on the GC cells. Importantly, Ntn4 level was significantly increased in 82 tumor tissues (p = 0.001) and 52 serum samples (p < 0.0001) from GC patients and positively correlated with Neo expression (p = 0.003). Ntn4 expression was negatively correlated with the survival period (p = 0.038), and positively associated with the severity of pathological stages of the tumors (p = 0.008). Taken together, Ntn4 promoted the proliferation and motility of GC cells which was mediated by its receptor Neo and through further activation of multi-oncogenic pathways. Elevated Ntn4 was detected in both tumor tissues and serum samples of GC patients and suggested a relatively poor survival, indicating Ntn4 may be used as a potential non-invasive biomarker for diagnosis and prognosis of GC.
netrin-4; gastric cancer; cell proliferation; cell motility
T helper- (Th-) cell immunodeficiency plays important roles in tumor development and their effects in chronic myeloid leukemia (CML) remain unclear. In the present study, we mainly investigated the role of Th22, Th17, and Th1 cell and their related cytokines (IL-22, IL-17, and IFN-r) in the pathophysiology of CML. Bone marrow (BM) and peripheral blood (PB) were extracted from newly diagnosed (ND), chronic phase- (CP-) CML patients, and controls. Th subsets were examined by flow cytometry. Plasma IL-22, IL-17, and IFN-r concentrations were measured by ELISA. AHR and RORC mRNA expressions were examined by RT-PCR. The frequencies of Th22, Th17, and Th1 cells, along with the expression of specific transcription factors RORC and AHR, were significantly decreased in ND patients compared with healthy controls, while all these abnormality recovered in CP patients. In addition, there existed a significantly positive relationship between Th22 and Th17 cells in PB or BM. A significantly negative relationship was found between Th cells (Th22, Th17, or Th1) and BCR-ABL (%) IS or the number of PB white blood cells. All these results demonstrated that Th22, Th17, and Th1 cells might be important therapeutic targets in CML and could facilitate a better outcome for tumor immunotherapy.
Liver cancer is the second-most frequent cause of cancer death in the world and is highly treatment resistant. We reported previously that inhibition of neddylation pathway with specific NAE inhibitor MLN4924, suppressed the malignant phenotypes of liver cancer. However, during the process, MLN4924 induces pro-survival autophagy as a mechanism of drug resistance. Here, we report that blockage of autophagy with clinically-available autophagy inhibitors (e.g. chloroquine) significantly enhanced the efficacy of MLN4924 on liver cancer cells by triggering apoptosis. Mechanistically, chloroquine enhanced MLN4924-induced up-regulation of pro-apoptotic proteins (e.g. NOXA) and down-regulation of anti-apoptotic proteins. Importantly, the down-regulation of NOXA expression via siRNA silencing substantially attenuated apoptosis of liver cancer cells. Further mechanistic studies revealed that blockage of autophagy augmented MLN4924-induced DNA damage and reactive oxygen species (ROS) generation. The elimination of DNA damage or blockage of ROS production significantly reduced the expression of NOXA, and thereby attenuated apoptosis and reduced growth inhibition of liver cancer cells. Moreover, blockage of autophagy enhanced the efficacy of MLN4924 in an orthotopic model of human liver cancer, with induction of NOXA and apoptosis in tumor tissues. These findings provide important preclinical evidence for clinical investigation of synergistic inhibition of neddylation and autophagy in liver cancer.
Neddylation; Autophagy; Apoptosis; MLN4924; Chloroquine
Gastric cancer is the fourth most common cancer worldwide, with a low 5-year survival rate. Epigenetic modification plays pivotal roles in gastric cancer development. However, the role of histone-modifying enzymes in gastric cancer remains largely unknown. Here we report that Sirt7, a NAD+-dependent class III histone deacetylase, is over-expressed in human gastric cancer tissues. Sirt7 level is significantly correlated with disease stage, metastasis, and survival. Knockdown of Sirt7 in gastric cancer cells inhibits cell proliferation and colony formation in vitro. In vivo subcutaneous xenograft results also show that Sirt7 knockdown can markedly repress gastric cancer cell growth. In addition, Sirt7 depletion induces apoptosis in gastric cancer cells via up-regulating expression of pro-apoptotic proteins and down-regulating anti-apoptotic proteins. Mechanically, Sirt7 binds to the promoter of miR-34a and deacetylases the H3K18ac, thus represses miR-34a expression. Reversely, depletion of miR-34a inhibits gastric cancer apoptosis induced by Sirt7 knockdown, and restores cellular capacity of proliferation and colony formation. miR-34a depletion reduces Sirt7-knockdown-induced arrest of gastric growth. Finally, miR-34a is tightly associated with survival of patients with gastric cancer.