ABSTRACT
BACKGROUND: The incidence of colorectal cancer and cancer death rate are increasing every year, and the affected population is becoming younger. Traditional Chinese medicine therapy has a unique effect in prolonging survival time and improving the prognosis of patients with colorectal cancer. Oridonin has been reported to have anti-cancer effects in a variety of tumors, but the exact mechanism remains to be investigated. METHODS: Cell Counting Kit-8 assay (CCK8) and 5-Ethynyl-2'-deoxyuridine (EdU) staining assay, Tranwell, and Wound healing assays were performed to measure cell proliferation, invasion, and migration capacities, respectively. The protein and mRNA expression levels of various molecules were reflected by Western blot and Reverse Transcription quantitative Polymerase Chain Reaction (qRT-PCR). Transcription Factor 4 (TCF4) and its target genes were analyzed by Position Weight Matrices (PWMs) software and the Gene Expression Omnibus (GEO) database. Immunofluorescence (IF) was performed to visualize the expression and position of Endoplasmic Reticulum (ER) stress biomarkers. The morphology of the ER was demonstrated by the ER tracker-red. Reactive Oxygen Species (ROS) levels were measured using a flow cytometer (FCM) or fluorescent staining. Calcium ion (Ca2+) concentration was quantified by Fluo-3 AM staining. Athymic nude mice were modeled with subcutaneous xenografts. RESULTS: Oridonin inhibited the proliferation, invasion, and migration of colorectal cancer, and this effect was weakened in a concentration-dependent manner by ER stress inhibitors. In addition, oridonin-induced colorectal tumor cells showed increased expression of ER stress biomarkers, loose morphology of ER, increased vesicles, and irregular shape. TCF4 was identified as a regulator of ER stress by PWMs software and GEO survival analysis. In vitro and in vivo experiments confirmed that TCF4 inhibited ER stress, reduced ROS production, and maintained Ca2+ homeostasis. In addition, oridonin also activated TP53 and inhibited TCF4 transactivation, further exacerbating the elevated ROS levels and calcium ion release in tumor cells and inhibiting tumorigenesis in colorectal cancer cells in vivo. CONCLUSIONS: Oridonin upregulated TP53, inhibited TCF4 transactivation, and induced ER stress dysregulation in tumor cells, promoting colorectal cancer cell death. Therefore, TCF4 may be one of the important nodes for tumor cells to regulate ER stress and maintain protein synthesis homeostasis. And the inhibition of the TP53/TCF4 axis plays a key role in the anti-cancer effects of oridonin.
Subject(s)
Apoptosis , Colorectal Neoplasms , Animals , Mice , Humans , Transcription Factor 4/genetics , Reactive Oxygen Species/metabolism , Calcium/metabolism , Mice, Nude , Transcriptional Activation , Cell Line, Tumor , Colorectal Neoplasms/drug therapy , Colorectal Neoplasms/genetics , Colorectal Neoplasms/metabolism , Endoplasmic Reticulum Stress , Cell Proliferation , Tumor Suppressor Protein p53/metabolismABSTRACT
Toosendanin (TSN) is the main active compound of Melia toosendan Sieb et Zucc with various bioactivities. In this study, we investigated the role of ferroptosis in TSN-induced hepatotoxicity. The characteristic indicators of ferroptosis were detected including reactive oxygen species (ROS), lipid-ROS, glutathione (GSH), ferrous ion and the expression of glutathione peroxidase 4 (GPX4), which showed that TSN caused ferroptosis in hepatocytes. The results of qPCR analysis and western blotting assay showed that TSN-induced activation of protein kinase R-like endoplasmic reticulum kinase (PERK)- eukaryotic initiation factor 2 α subunit (eIF2α)- activation transcription factor 4 (ATF4) signaling pathway resulted in increasing activation transcription factor 3 (ATF3) expression, which upregulated the expression of transferrin receptor 1 (TFRC). Furthermore, TFRC mediated iron accumulation leading to ferroptosis in hepatocytes. To clarify whether TSN triggered ferroptosis in vivo, male Balb/c mice were treated with the different doses of TSN. The results of hematoxylin-eosin (H&E) staining, 4-hydroxynonenal (4-HNE) staining, malondialdehyde (MDA) content and the protein expression of GPX4 showed that ferroptosis contributed to TSN-induced hepatotoxicity. Iron homeostasis relative protein and PERK- eIF2α- ATF4 signaling pathway also involved in hepatotoxicity of TSN in vivo.
Subject(s)
Chemical and Drug Induced Liver Injury , Drugs, Chinese Herbal , Ferroptosis , Animals , Mice , Male , Eukaryotic Initiation Factor-2/metabolism , Reactive Oxygen Species/metabolism , Transcription Factor 4 , Activating Transcription Factor 4/genetics , Activating Transcription Factor 4/metabolismABSTRACT
Laryngeal cancer (LC) is a rare and challenging clinical problem. Our aim was to investigate the mechanism of salt-like transcription factor 4 (SALL4) in LC. LC tissue and paracancerous tissue were collected. Relative mRNA or protein levels were measured by quantitative real-time polymerase chain reaction or Western blot. MTT, wound healing, and transwell assay were performed to evaluate cell proliferation, migration and invasion. The binding relationship between SALL4 and USP21 promoter was verified by dual-luciferase assay and ChIP. Co-IP and glutathione-S-transferase (GST)-pull down were performed to measure the protein interaction between USP21 and YY1. Additionally, YY1 ubiquitination level was analyzed. It was found that SALL4 mRNA and SALL4 protein levels were elevated in LC clinical tissues and various LC cells. Knockdown of SALL4 inhibited epithelial-mesenchymal transition (EMT) of LC cells. USP21 was transcriptionally activated by SALL4. Co-IP and GST-pull down confirmed USP21 interacted with YY1. USP21 protected YY1 from degradation through deubiquitination. Furthermore, overexpression of USP21 reversed the effect of knockdown of SALL4 on YY1 and EMT in LC cells. In general, SALL4 facilitated EMT of LC cells through modulating USP21/YY1 axis.
Subject(s)
Laryngeal Neoplasms , Transcription Factors , Humans , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation/genetics , Gene Expression Regulation, Neoplastic , Laryngeal Neoplasms/genetics , RNA, Messenger , Transcription Factor 4/genetics , Transcription Factor 4/metabolism , Transcription Factors/genetics , Transcription Factors/metabolism , Transcriptional Activation , Ubiquitin Thiolesterase/genetics , Ubiquitin-Specific Proteases/genetics , Ubiquitin-Specific Proteases/metabolism , Yin-YangABSTRACT
Cachexia, which is characterised by the wasting of fat and skeletal muscles, is the most common risk factor for increased mortality rates among patients with advanced lung cancer. PTHLH (parathyroid hormone-like hormone) is reported to be involved in the pathogenesis of cancer cachexia. However, the molecular mechanisms underlying the regulation of PTHLH expression and the inhibitors of PTHLH have not yet been identified. The PTHLH mRNA levels were measured using quantitative real-time polymerase chain reaction, while the PTHrP (parathyroid hormone-related protein) expression levels were measured using Western blotting and enzyme-linked immunosorbent assay. The interaction between TCF4 (Transcription Factor 4) and TWIST1 and the binding of the TCF4-TWIST1 complex to the PTHLH promoter were analysed using co-immunoprecipitation and chromatin immunoprecipitation. The results of the mammalian two-hybrid luciferase assay revealed that emodin inhibited TCF4-TWIST1 interaction. The effects of Polygonum cuspidatum extract (Pc-Ex), which contains emodin, on cachexia were investigated in vivo using A549 tumour-bearing mice. Ectopic expression of TCF4 upregulated PTHLH expression. Conversely, TCF4 knockdown downregulated PTHLH expression in lung cancer cells. The expression of PTHLH was upregulated in cells ectopically co-expressing TCF4 and TWIST1 when compared with that in cells expressing TCF4 or TWIST1 alone. Emodin inhibited the interaction between TCF4 and TWIST1 and consequently suppressed the TCF4/TWIST1 complex-induced upregulated mRNA and protein levels of PTHLH and PTHrP. Meanwhile, emodin-containing Pc-Ex significantly alleviated skeletal muscle atrophy and downregulated fat browning-related genes in A549 tumour-bearing mice. Emodin-containing Pc-Ex exerted therapeutic effects on lung cancer-associated cachexia by inhibiting TCF4/TWIST1 complex-induced PTHrP expression.
Subject(s)
Emodin , Fallopia japonica , Lung Neoplasms , Animals , Cachexia/drug therapy , Cachexia/etiology , Cachexia/prevention & control , Emodin/pharmacology , Emodin/therapeutic use , Humans , Lung Neoplasms/complications , Lung Neoplasms/drug therapy , Mammals/genetics , Mammals/metabolism , Mice , Nuclear Proteins/genetics , Parathyroid Hormone-Related Protein/genetics , Plant Extracts , RNA, Messenger/metabolism , Transcription Factor 4/genetics , Twist-Related Protein 1/geneticsABSTRACT
Neuronal phenotypes are controlled by terminal selector transcription factors in invertebrates, but only a few examples of such regulators have been provided in vertebrates. We hypothesised that TCF7L2 regulates different stages of postmitotic differentiation in the thalamus, and functions as a thalamic terminal selector. To investigate this hypothesis, we used complete and conditional knockouts of Tcf7l2 in mice. The connectivity and clustering of neurons were disrupted in the thalamo-habenular region in Tcf7l2-/- embryos. The expression of subregional thalamic and habenular transcription factors was lost and region-specific cell migration and axon guidance genes were downregulated. In mice with a postnatal Tcf7l2 knockout, the induction of genes that confer thalamic terminal electrophysiological features was impaired. Many of these genes proved to be direct targets of TCF7L2. The role of TCF7L2 in terminal selection was functionally confirmed by impaired firing modes in thalamic neurons in the mutant mice. These data corroborate the existence of master regulators in the vertebrate brain that control stage-specific genetic programmes and regional subroutines, maintain regional transcriptional network during embryonic development, and induce terminal selection postnatally.
Subject(s)
Cell Differentiation , Gene Expression Regulation, Developmental , Mitosis , Synaptic Transmission , Thalamus/embryology , Transcription Factor 4/metabolism , Animals , Mice , Mice, Knockout , Thalamus/cytology , Transcription Factor 4/geneticsABSTRACT
The voltage sodium channel 1.8 (NaV1.8) in the dorsal root ganglion (DRG) neurons contributes to the initiation and development of chronic inflammatory and neuropathic pain. However, an effective intervention on NaV1.8 remains to be studied in pre-clinical research and clinical trials. In this study, we aimed to investigate whether transcription factor 4 (TCF4) overexpression represses NaV1.8 expression in DRG neurons, thus preventing the development of chronic pain. Using chromatin immunoprecipitation (CHIP), we verified the interaction of TCF4 and sodium voltage-gated channel alpha subunit 10A (SCN10A) enhancer in HEK293 cells and rat DRG neurons. Using a dual luciferase reporter assay, we confirmed the transcriptional inhibition of TCF4 on SCN10A promoter in vitro. To investigate the regulation of TCF4 on Nav1.8, we then upregulated TCF4 expression by intrathecally delivering an overexpression of recombinant adeno-associated virus (rAAV) in the Complete Freund's adjuvant (CFA)-induced inflammatory pain model and spared nerve injury (SNI)-induced neuropathic pain model. By using a quantitative polymerase chain reaction (qPCR), western blot, and immunostaining, we evaluated NaV1.8 expression after a noxious stimulation and the application of the TCF4 overexpression virus. We showed that the intrathecal delivery of TCF4 overexpression virus significantly repressed the increase of NaV1.8 and prevented the development of hyperalgesia in rats. Moreover, we confirmed the efficient role of an overexpressed TCF4 in preventing the CFA- and SNI-induced neuronal hyperexcitability by calcium imaging. Our results suggest that attenuating the dysregulation of NaV1.8 by targeting TCF4 may be a novel therapeutic strategy for chronic inflammatory and neuropathic pain.
Subject(s)
Ganglia, Spinal/metabolism , Hyperalgesia/metabolism , NAV1.8 Voltage-Gated Sodium Channel/metabolism , Neuralgia/metabolism , Neurons/metabolism , Transcription Factor 4/metabolism , Animals , Down-Regulation , HEK293 Cells , Humans , Hyperalgesia/genetics , Inflammation/genetics , Inflammation/metabolism , Male , NAV1.8 Voltage-Gated Sodium Channel/genetics , Neuralgia/genetics , Pain Threshold/physiology , Rats , Rats, Sprague-Dawley , Transcription Factor 4/genetics , Up-RegulationABSTRACT
This study aimed to investigate the potential therapeutic targets of Liuwei Dihuang pill (LDP) in the treatment of postmenopausal osteoporosis with kidney-Yin deficiency (PMO-KY).Gene expression data were downloaded from the GEO database, including 4 PMO-KY samples and 3 healthy postmenopausal controls from GSE56116, as well as 3 PMO-KY samples before LDP treatment and 3 PMO-KY samples after three months of LDP treatment from GSE57273. Limma package was used to identify differentially expressed genes (DEGs). Afterwards, the potential target genes of LDP (namely key DEGs) were identified according to the comparison of DEGs in PMO-KY group and the DEGs in LDP treatment groups. Subsequently, iRegulon plugin in Cytoscape software was used to predict potential transcription factors (TFs) that regulated the key DEGs, and Comparative Toxicogenomics Database was utilized to identify known PMO-related genes among the key DEGs.Totally, 202 and 2066 DEGs were identified between PMO-KY and controls, as well as after-treatment and before-treatment groups, respectively. Among them, 52 DEGs were up-regulated in PMO-KY but down-regulated after LDP treatment, and 8 TFs were predicted to these DEGs. Furthermore, 34 DEGs were down-regulated in PMO-KY but up-regulated after treatment, and 7 TFs were predicted to regulate these DEGs. Additionally, 43 of the 86 key DEGs were known PMO-related genes.NCOA3, TCF4, DUSP6, PELI2, and STX7 were predicted to be regulated by HOXA13. In the PMO-KY treatment, NCOA3, TCF4, DUSP6, PELI2, and STX7 might be the potential therapeutic targets of LDP. However, further investigation is required to confirm these genes.
Subject(s)
Drugs, Chinese Herbal/pharmacology , Osteoporosis, Postmenopausal/drug therapy , Osteoporosis, Postmenopausal/genetics , Yin Deficiency/drug therapy , Yin Deficiency/genetics , Case-Control Studies , Dual Specificity Phosphatase 6/drug effects , Female , Gene Expression Profiling , Homeodomain Proteins/drug effects , Humans , Kidney/metabolism , Middle Aged , Nuclear Proteins/drug effects , Nuclear Receptor Coactivator 3/drug effects , Qa-SNARE Proteins/drug effects , Toxicogenetics , Transcription Factor 4/drug effects , Ubiquitin-Protein Ligases/drug effectsABSTRACT
Sensorimotor gating measured by prepulse inhibition (PPI) of the acoustic startle response (ASR) has been proposed as one of the most promising electrophysiological endophenotypes of schizophrenia. During the past decade, a number of publications have reported significant associations between genetic polymorphisms and PPI in samples of schizophrenia patients and healthy volunteers. However, an overall evaluation of the robustness of these results has not been published so far. Therefore, we performed the first meta-analysis of published and unpublished associations between gene polymorphisms and PPI of ASR. Unpublished associations between genetic polymorphisms and PPI were derived from three independent samples. In total, 120 single observations from 16 independent samples with 2660 study participants and 43 polymorphisms were included. After correction for multiple testing based on false discovery rate and considering the number of analyzed polymorphisms, significant associations were shown for four variants, even though none of these associations survived a genome-wide correction (P<5∗10-8). These results imply that PPI might be modulated by four genotypes - COMT rs4680 (primarily in males), GRIK3 rs1027599, TCF4 rs9960767, and PRODH rs385440 - indicating a role of these gene variations in the development of early information processing deficits in schizophrenia. However, the overall impact of single genes on PPI is still rather small suggesting that PPI is - like the disease phenotype - highly polygenic. Future genome-wide analyses studies with large sample sizes will enhance our understanding on the genetic architecture of PPI.
Subject(s)
Acoustic Stimulation , Gait Disorders, Neurologic/genetics , Polymorphism, Genetic/genetics , Reflex, Startle/genetics , Catechol O-Methyltransferase/genetics , Gait Disorders, Neurologic/etiology , Genome-Wide Association Study , Humans , Proline Oxidase/genetics , Receptors, Kainic Acid/genetics , Schizophrenia/complications , Transcription Factor 4/genetics , GluK3 Kainate ReceptorABSTRACT
A strategy to suppress the expression of the DNA repair enzyme O6methylguanineDNA methyltransferase (MGMT) by inhibition of Wnt/ßcatenin signaling may be useful as a novel treatment for pituitary adenoma. Previous studies have reported that Tanshinone IIA (TSA), a major quinone compound isolated from Salvia miltiorrhiza, had antitumor effects. However, whether TSA has antitumor effects against pituitary adenoma and whether the mechanisms are associated with the Wnt/ßcatenin/MGMT pathway remains to be clarified. In the present study, TSA treatment caused apoptosis in AtT20 cells in a concentrationdependent manner, as demonstrated by cell viability reduction, phophatidylserine externalization detected by Annexin V staining and mitochondrial membrane potential disruption detected by JC1 staining, which were associated with activation of caspase3 and DNA fragmentation detected by TUNEL in AtT20 cells. Tcell factor (TCF)lymphoidenhancing factor (LEF) reporter activity was determined by dual luciferase reporter assay and the interaction between ßcatenin and TCF4 were detected using a coimmunoprecipitation kit. The results indicated TSA treatment increased ßcatenin phosphorylation, inhibited ßcatenin nuclear translocation, reduced ßcatenin/TCF4 complex formation and TCFLEF luciferase reporter activity, and subsequently reduced the expression of cyclin D1 and MGMT. Notably, overexpression of MGMT in ßcatenin knock down AtT20 cells abrogated the TSAmediated effects in AtT20 cells. In conclusion, TSA induced apoptosis via inhibition of Wnt/ßcatenindependent MGMT expression, which may provide novel insights into the understanding of the mechanism of the antitumor effects of Salvia miltiorrhiza.
Subject(s)
Abietanes/pharmacology , Antineoplastic Agents, Phytogenic/pharmacology , Corticotrophs/drug effects , DNA Modification Methylases/genetics , DNA Repair Enzymes/genetics , Gene Expression Regulation, Neoplastic , Salvia miltiorrhiza/chemistry , Tumor Suppressor Proteins/genetics , beta Catenin/genetics , Abietanes/isolation & purification , Animals , Antineoplastic Agents, Phytogenic/isolation & purification , Apoptosis/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Corticotrophs/metabolism , Corticotrophs/pathology , DNA Fragmentation , DNA Modification Methylases/antagonists & inhibitors , DNA Modification Methylases/metabolism , DNA Repair Enzymes/antagonists & inhibitors , DNA Repair Enzymes/metabolism , Dose-Response Relationship, Drug , Humans , Lymphoid Enhancer-Binding Factor 1/genetics , Lymphoid Enhancer-Binding Factor 1/metabolism , Mice , Plant Extracts/chemistry , RNA, Small Interfering/genetics , RNA, Small Interfering/metabolism , Transcription Factor 4/genetics , Transcription Factor 4/metabolism , Tumor Suppressor Proteins/antagonists & inhibitors , Tumor Suppressor Proteins/metabolism , Wnt Signaling Pathway/drug effects , beta Catenin/antagonists & inhibitors , beta Catenin/metabolismABSTRACT
Environmental enrichment (EE) replicates mind-body therapy by providing complex housing to laboratory animals to improve their activity levels, behavior, and social interactions. Using a Tcf4Het/+ApcMin/+-mediated model of colon tumorigenesis, we found that EE vastly improved the survival of tumor-bearing animals, with differential effect on tumor load in male compared to female animals. Analysis of Tcf4Het/+ApcMin/+ males showed drastically reduced expression of circulating inflammatory cytokines and induced nuclear hormone receptor (NHR) signaling, both of which are common in the wound repair process. Interestingly, EE provoked tumor wound repair resolution through revascularization, plasma cell recruitment and IgA secretion, replacement of glandular tumor structures with pericytes in a process reminiscent of scarring, and normalization of microbiota. These EE-dependent changes likely underlie the profound improvement in survival of colon-tumor-bearing Tcf4Het/+ApcMin/+ males. Our studies highlight the exciting promise of EE in the design of future therapeutic strategies for colon cancer patients.
Subject(s)
Colonic Neoplasms/pathology , Environment , Immunoglobulin A/metabolism , Wound Healing/physiology , Adenomatous Polyposis Coli Protein/genetics , Adenomatous Polyposis Coli Protein/metabolism , Alphaproteobacteria/isolation & purification , Alphaproteobacteria/physiology , Animals , Colon/microbiology , Colonic Neoplasms/metabolism , Colonic Neoplasms/mortality , Cytokines/blood , Disease Models, Animal , Female , Male , Mice , Microbiota , Neovascularization, Physiologic , Pericytes/cytology , Pericytes/metabolism , Proteobacteria/isolation & purification , Proteobacteria/physiology , Receptors, Cytoplasmic and Nuclear/metabolism , Signal Transduction , Survival Rate , Transcription Factor 4/genetics , Transcription Factor 4/metabolismABSTRACT
Berberine chloride (BBR) is an isoquinoline derivative alkaloid isolated from medicinal herbs, including Coptis chinensis and Berberis aristate. This compound plays significant roles in the treatment of osteoarthritis (OA). The purpose of this study was to investigate the effects of BBR on the proliferation of sodium nitroprusside (SNP)-stimulated chondrocytes in vitro, the articular cartilage in a rat OA model, as well as to discuss the molecular mechanisms underlying these effects. In vitro, we demonstrated that BBR led to cell proliferation, increased the cell population in S-phase and decreased that in G0/G1-phase; moreover, the F-actin remodeling in SNP-stimulated chondrocytes were prevented. In addition, BBR markedly up-regulated ß-catenin, c-Myc, and cyclin D1 expression of genes and proteins, and down-regulated glycogen synthase kinase-3ß (GSK-3ß) and matrix metalloproteinase-7 (MMP-7) expression. Notably, inhibition of the Wnt/ß-catenin pathway by XAV939 partially blocked these effects. The in vivo results suggested that BBR promoted ß-catenin protein level and enhanced proliferating cell nuclear antigen (PCNA) expression in osteoarthritic rat cartilage. In conclusion, these findings indicate that BBR promotes SNP-stimulated chondrocyte proliferation by promoting G1/S phase transition and synthesis of PCNA in cartilage through activation of Wnt/ß-catenin signaling pathway.
Subject(s)
Berberine/pharmacology , Cartilage, Articular/pathology , Chondrocytes/cytology , Chondrocytes/drug effects , Nitroprusside/adverse effects , Osteoarthritis/pathology , Wnt Signaling Pathway/drug effects , Animals , Cell Cycle/drug effects , Cell Proliferation/drug effects , Chondrocytes/metabolism , Chondrocytes/pathology , Cyclin D1/genetics , Cytoskeleton/drug effects , Cytoskeleton/metabolism , DNA-Binding Proteins/genetics , Gene Expression Regulation, Enzymologic/drug effects , Glycogen Synthase Kinase 3 beta/genetics , Lymphoid Enhancer-Binding Factor 1/metabolism , Proliferating Cell Nuclear Antigen/metabolism , Proto-Oncogene Proteins c-myc/genetics , Rats , Rats, Sprague-Dawley , Transcription Factor 4 , Transcription Factors/geneticsABSTRACT
Previous research has shown that total saponins of Panax ginseng (TSPG) and other ginsenoside monomers inhibit the proliferation of leukemia cells. However, the effect has not been compared among them. Cell viability was determined by Cell Counting Kit-8 assay, and ultra-structural characteristics were observed under transmission electron microscopy. Cell cycle distribution and apoptosis were determined by flow cytometry (FCM). Real-time fluorescence quantitativePCR, western blotting and immunofluorescence were used to measure the expression of ß-catenin, TCF4, cyclin D1 and NF-κBp65. ß-catenin/TCF4 target gene transcription were observed by ChIP-PCR assay. We found that 20(S)-ginsenoside Rh2 [(S)Rh2] inhibited the proliferation of KG-1a cells more efficiently than the other monomers. Moreover, (S)Rh2 arrested KG-1a cells in the G0/G1 phase and induced apoptosis. In addition, the levels of ß-catenin, TCF4, cyclin D1 mRNA and protein were decreased. The ChIP-PCR showed that (S)Rh2 downregulated the transcription of ß-catenin/TCF4 target genes, such as cyclin D1 and c-myc. These results indicated that (S)Rh2 induced cell cycle arrest and apoptosis through the Wnt/ß-catenin signaling pathway, demonstrating its potential as a chemotherapeutic agent for leukemia therapy.
Subject(s)
Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Cell Proliferation/drug effects , Ginsenosides/pharmacology , Leukemia/drug therapy , Wnt Signaling Pathway/drug effects , Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/metabolism , Cell Line, Tumor , Cyclin D1/metabolism , Drugs, Chinese Herbal/pharmacology , Flow Cytometry , G1 Phase Cell Cycle Checkpoints/drug effects , Humans , Leukemia/pathology , Microscopy, Electron, Transmission , Panax/metabolism , Real-Time Polymerase Chain Reaction , Transcription Factor 4 , Transcription Factor RelA/metabolism , Transcription Factors/metabolism , Wnt Proteins/metabolism , beta Catenin/metabolismABSTRACT
Schizophrenia (SZ) is a severe mental disorder affecting about 1 % of the human population. Patients show severe deficits in cognitive processing often characterized by an improper filtering of environmental stimuli. Independent genome-wide association studies confirmed a number of risk variants for SZ including several associated with the gene encoding the transcription factor 4 (TCF4). TCF4 is widely expressed in the central nervous system of mice and humans and seems to be important for brain development. Transgenic mice overexpressing murine Tcf4 (Tcf4tg) in the adult brain display cognitive impairments and sensorimotor gating disturbances. To address the question of whether increased Tcf4 gene dosage may affect cognitive flexibility in an auditory associative task, we tested latent inhibition (LI) in female Tcf4tg mice. LI is a widely accepted translational endophenotype of SZ and results from a maladaptive delay in switching a response to a previously unconditioned stimulus when this becomes conditioned. Using an Audiobox, we pre-exposed Tcf4tg mice and their wild-type littermates to either a 3- or a 12-kHz tone before conditioning them to a 12-kHz tone. Tcf4tg animals pre-exposed to a 12-kHz tone showed significantly delayed conditioning when the previously unconditioned tone became associated with an air puff. These results support findings that associate TCF4 dysfunction with cognitive inflexibility and improper filtering of sensory stimuli observed in SZ patients.
Subject(s)
Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/genetics , Cognition Disorders/genetics , General Adaptation Syndrome/genetics , Sensory Gating/genetics , Acoustic Stimulation , Animals , Disease Models, Animal , Exploratory Behavior/physiology , Female , Inhibition, Psychological , Locomotion/genetics , Mice , Mice, Inbred C57BL , Mice, Transgenic , Psychoacoustics , Transcription Factor 4ABSTRACT
There is mounting evidence that noncoding microRNAs (miRNA) are modulated by select chemoprotective dietary agents. For example, recently we demonstrated that the unique combination of dietary fish oil (containing n-3 fatty acids) plus pectin (fermented to butyrate in the colon) (FPA) up-regulates a subset of putative tumor suppressor miRNAs in intestinal mucosa, and down-regulates their predicted target genes following carcinogen exposure as compared to control (corn oil plus cellulose (CCA)) diet. To further elucidate the biological effects of diet and carcinogen modulated miR's in the colon, we verified that miR-26b and miR-203 directly target PDE4B and TCF4, respectively. Since perturbations in adult stem cell dynamics are generally believed to represent an early step in colon tumorigenesis and to better understand how the colonic stem cell population responds to environmental factors such as diet and carcinogen, we additionally determined the effects of the chemoprotective FPA diet on miRNAs and mRNAs in colonic stem cells obtained from Lgr5-EGFP-IRES-creER(T2) knock-in mice. Following global miRNA profiling, 26 miRNAs (P<0.05) were differentially expressed in Lgr5(high) stem cells as compared to Lgr5(negative) differentiated cells. FPA treatment up-regulated miR-19b, miR-26b and miR-203 expression as compared to CCA specifically in Lgr5(high) cells. In contrast, in Lgr5(negative) cells, only miR-19b and its indirect target PTK2B were modulated by the FPA diet. These data indicate for the first time that select dietary cues can impact stem cell regulatory networks, in part, by modulating the steady-state levels of miRNAs. To our knowledge, this is the first study to utilize Lgr5(+) reporter mice to determine the impact of diet and carcinogen on miRNA expression in colonic stem cells and their progeny.
Subject(s)
Carcinogens , Colon/pathology , Colonic Neoplasms/genetics , Diet , Fatty Acids, Omega-3/metabolism , Gene Expression Regulation, Neoplastic , MicroRNAs/genetics , Stem Cell Niche , Animals , Carcinogens/metabolism , Carcinogens/toxicity , Colon/metabolism , Colonic Neoplasms/etiology , Colonic Neoplasms/pathology , Cyclic Nucleotide Phosphodiesterases, Type 4/genetics , Focal Adhesion Kinase 2/genetics , Gene Expression Regulation, Neoplastic/drug effects , HCT116 Cells , Humans , Mice , Protective Factors , Stem Cell Niche/drug effects , Transcription Factor 4/geneticsABSTRACT
The PI3K-Akt-mTOR, Wnt/ß-catenin and apoptosis signaling pathways have been shown to be involved in genesis of colorectal cancer (CRC). The aim of this study was to elucidate whether combination of Gelam honey and ginger might have chemopreventive properties in HT29 colon cancer cells by modulating the mTOR, Wnt/ß-catenin and apoptosis signaling pathways. Treatment with Gelam honey and ginger reduced the viability of the HT29 cells dose dependently with IC50 values of 88 mg/ml and 2.15 mg/ml respectively, their while the combined treatment of 2 mg/ml of ginger with 31 mg/ml of Gelam honey inhibited growth of most HT29 cells. Gelam honey, ginger and combination induced apoptosis in a dose dependent manner with the combined treatment exhibiting the highest apoptosis rate. The combined treatment downregulated the gene expressions of Akt, mTOR, Raptor, Rictor, ß-catenin, Gsk3ß, Tcf4 and cyclin D1 while cytochrome C and caspase 3 genes were shown to be upregulated. In conclusion, the combination of Gelam honey and ginger may serve as a potential therapy in the treatment of colorectal cancer through inhibiton of mTOR, Wnt/ß catenin signaling pathways and induction of apoptosis pathway.
Subject(s)
Anticarcinogenic Agents/pharmacology , Honey , Plant Extracts/pharmacology , TOR Serine-Threonine Kinases/genetics , Wnt Signaling Pathway/drug effects , Zingiber officinale , Adaptor Proteins, Signal Transducing/genetics , Apoptosis/drug effects , Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/genetics , Carrier Proteins/genetics , Caspase 3/genetics , Cell Survival/drug effects , Chemoprevention , Cyclin D1/genetics , Cytochromes c/genetics , Dose-Response Relationship, Drug , Down-Regulation/drug effects , Gene Expression/drug effects , Glycogen Synthase Kinase 3/genetics , Glycogen Synthase Kinase 3 beta , HT29 Cells , Humans , Inhibitory Concentration 50 , Melaleuca , Proto-Oncogene Proteins c-akt/genetics , Rapamycin-Insensitive Companion of mTOR Protein , Regulatory-Associated Protein of mTOR , TOR Serine-Threonine Kinases/metabolism , Transcription Factor 4 , Transcription Factors/genetics , Up-Regulation/drug effects , beta Catenin/geneticsABSTRACT
The powerful genome-wide association studies (GWAS) revealed common mutations that increase susceptibility for schizophrenia (SZ) and bipolar disorder (BD), but the vast majority were not known to be functional or associated with these illnesses. To help fill this gap, their impact on human brain structure and function has been examined. We systematically discuss this output to facilitate its timely integration in the psychosis research field; and encourage reflection for future research. Irrespective of imaging modality, studies addressing the effect of SZ/BD GWAS risk genes (ANK3, CACNA1C, MHC, TCF4, NRGN, DGKH, PBRM1, NCAN and ZNF804A) were included. Most GWAS risk variations were reported to affect neuroimaging phenotypes implicated in SZ/BD: white-matter integrity (ANK3 and ZNF804A), volume (CACNA1C and ZNF804A) and density (ZNF804A); grey-matter (CACNA1C, NRGN, TCF4 and ZNF804A) and ventricular (TCF4) volume; cortical folding (NCAN) and thickness (ZNF804A); regional activation during executive tasks (ANK3, CACNA1C, DGKH, NRGN and ZNF804A) and functional connectivity during executive tasks (CACNA1C and ZNF804A), facial affect recognition (CACNA1C and ZNF804A) and theory-of-mind (ZNF804A); but inconsistencies and non-replications also exist. Further efforts such as standardizing reporting and exploring complementary designs, are warranted to test the reproducibility of these early findings.
Subject(s)
Bipolar Disorder/genetics , Bipolar Disorder/physiopathology , Brain/physiopathology , Genetic Predisposition to Disease , Schizophrenia/genetics , Schizophrenia/physiopathology , Ankyrins/genetics , Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/genetics , Bipolar Disorder/psychology , Calcium Channels, L-Type/genetics , Chondroitin Sulfate Proteoglycans/genetics , Female , Functional Neuroimaging , Genes , Genetic Predisposition to Disease/genetics , Genetic Predisposition to Disease/psychology , Genome , Genome-Wide Association Study , Humans , Kruppel-Like Transcription Factors/genetics , Lectins, C-Type/genetics , Male , Mutation/genetics , Nerve Tissue Proteins/genetics , Neurocan , Neurogranin/genetics , Phenotype , Polymorphism, Genetic , Psychotic Disorders/physiopathology , Risk Factors , Schizophrenic Psychology , Transcription Factor 4 , Transcription Factors/geneticsABSTRACT
OBJECTIVE: To study the effect of Biejiajian Pills on Wnt signal pathway and the mechanisms underlying its action to suppress the invasiveness of hepatocellular carcinoma. METHODS: HepG2 cells cultured in the serum of rats fed with Biejiajian Pills for 48 h were examined for ß-catenin expression using immunofluorescence, ß-catenin/TCF4 complex activity with luciferase, and expressions of the downstream proteins cyclin D1 and MMP-2 using qRT-PCR. RESULTS: Biejiajian Pills-treated sera significantly reduced the expressions of cytoplasmic and nuclear ß-catenin protein, cyclin D1 and MMP-2 proteins and lowered the activities of ß-catenin/TCF4 complex. CONCLUSION: Biejiajian Pills may serve as a potential anti-tumor agent, whose effect might be mediated by inhibiting the Wnt/ß-catenin pathway.
Subject(s)
Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/metabolism , Cyclin D1/metabolism , Drugs, Chinese Herbal/pharmacology , Matrix Metalloproteinase 2/metabolism , Transcription Factors/metabolism , Wnt Signaling Pathway , beta Catenin/metabolism , Animals , Carcinoma, Hepatocellular/metabolism , Hep G2 Cells , Humans , Liver Neoplasms/metabolism , Rats , Transcription Factor 4 , Wnt ProteinsABSTRACT
Sweet potato leaves contain the highest levels of functional polyphenols. In this study the effects of the sweet potato leaf extract and its contents, such as mono (3, 4, and 5)-caffeoylquinic acid (CQA), di-CQA (4,5-diCQA, 3,5-diCQA, and 3,4-diCQA) and caffeic acid (CA), were evaluated on the ß-catenin/Tcf-4 signaling in human colorectal cancer HCT116 cells. The extract and the CQA derivatives inhibited the ß-catenin/Tcf-4 signaling, and the inhibition of the di-CQA (with two caffeoyl groups) was higher than that of the mono-CQA (one-caffeoyl group) and CA, suggesting that the caffeoyl structure in the presence of a catechol group plays a significant role in interfering with the ß-catenin/Tcf-4 signaling. In addition, the CQA derivatives had no effect on the ß-catenin protein expression, but all test compounds inhibited the expression of the Tcf-4 transcription, and the inhibition of the di-CQA derivatives was stronger than those of the mono-CQA derivatives as well as the ß-catenin/Tcf-4 transcriptional activity. These compounds can modulate the downstream Wnt signaling pathway, suggesting that sweet potato leaves can be a protective food for colorectal cancer.
Subject(s)
Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/genetics , Down-Regulation/drug effects , Ipomoea batatas/chemistry , Plant Extracts/pharmacology , Quinic Acid/analogs & derivatives , Transcription Factors/genetics , beta Catenin/metabolism , Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/metabolism , HCT116 Cells , Humans , Plant Leaves/chemistry , Quinic Acid/pharmacology , Signal Transduction/drug effects , Transcription Factor 4 , Transcription Factors/metabolismABSTRACT
<p><b>OBJECTIVE</b>To study the effect of Biejiajian Pills on Wnt signal pathway and the mechanisms underlying its action to suppress the invasiveness of hepatocellular carcinoma.</p><p><b>METHODS</b>HepG2 cells cultured in the serum of rats fed with Biejiajian Pills for 48 h were examined for β-catenin expression using immunofluorescence, β-catenin/TCF4 complex activity with luciferase, and expressions of the downstream proteins cyclin D1 and MMP-2 using qRT-PCR.</p><p><b>RESULTS</b>Biejiajian Pills-treated sera significantly reduced the expressions of cytoplasmic and nuclear β-catenin protein, cyclin D1 and MMP-2 proteins and lowered the activities of β-catenin/TCF4 complex.</p><p><b>CONCLUSION</b>Biejiajian Pills may serve as a potential anti-tumor agent, whose effect might be mediated by inhibiting the Wnt/β-catenin pathway.</p>
Subject(s)
Animals , Humans , Rats , Basic Helix-Loop-Helix Leucine Zipper Transcription Factors , Metabolism , Carcinoma, Hepatocellular , Metabolism , Cyclin D1 , Metabolism , Drugs, Chinese Herbal , Pharmacology , Hep G2 Cells , Liver Neoplasms , Metabolism , Matrix Metalloproteinase 2 , Metabolism , Transcription Factor 4 , Transcription Factors , Metabolism , Wnt Proteins , Wnt Signaling Pathway , beta Catenin , MetabolismABSTRACT
BACKGROUND: z-Guggulsterone (z-Gug) and Gugulipid (GL) have been used to treat a variety of ailments. We now report their anti-cancer effect and mechanism against human breast cancer. METHODS: Using the human estrogen receptor-positive (MCF-7) and triple-negative (MDA-MB-231) breast cancer cells as well as the normal human mammary epithelial cell line (HMEC), we evaluated the anti-breast-cancer efficacy and apoptosis inducing activity of GL. We determined the cellular and molecular mechanism of GL-inhibited breast cancer cell growth. RESULTS: GL significantly inhibited growth of MCF-7 and MDA-MB-231 cells with an IC50~2 µM at pharmacologically relevant concentrations standardized to its major active constituent z-Gug. The GL-induced growth inhibition correlated with apoptosis induction as evidenced by an increase in cytoplasmic histone-associated DNA fragmentation and caspase 3 activity. The GL-induced apoptosis was associated with down-regulation of the ß-Catenin signaling pathway. The decreased expression of Wnt/ß-Catenin targeting genes, such as cyclin D1, C-myc and survivin, and the inhibition of the activity of the transcription factor (T-cell factor 4, TCF-4) were observed in GL-treated breast cancer cells. The GL treatment resulted in a significant reduction of ß-Catenin /TCF-4 complex in both of the cancer cells. The GL-induced apoptotic cell death was significantly enhanced by RNA Interference of ß-Catenin and TCF-4. On the other hand, the normal human mammary epithelial cell HMEC, compared with the human breast cancer cells, is significantly more resistant to growth inhibition and apoptosis induction by GL. CONCLUSION: The present study indicates that the ß-Catenin signaling pathway is the target for GL-induced growth inhibition and apoptosis in human breast cancer.