ABSTRACT
Apoptosis signal-regulated kinase 1 (ASK1) is a member of the mitogen-activated protein kinase kinase (MAP3K) family, whose activation and regulation are intricately associated with apoptosis. ASK1 is activated in response to oxidative stress, among other stimuli, subsequently triggering downstream JNK, p38 MAPK, and mitochondria-dependent apoptotic signaling, which participate in the initiation of tumor cell apoptosis induced by various stimuli. Research has shown that ASK1 plays a crucial role in the apoptosis of lung cancer, breast cancer, and liver cancer cells. Currently, the investigation of effective ASK1 activators is a hot topic in research on tumor cell apoptosis. Synthetic compounds such as human ß-defensin, triazolothiazide derivatives and heat shock protein 27 inhibitors; natural compounds such as quercetin, Laminarina japonica polysaccharide-1 peptide and theabrownin; and nanomedicines such as cerium oxide nanoparticles, magnetite FeO nanoparticles and silver nanoparticles can activate ASK1 and induce apoptosis in various tumor cells. This review extensively investigates the roles and activation mechanisms of ASK1, explores its impact on a variety of apoptotic signaling pathways, and discusses the potential therapeutic applications of various ASK1 activators in cancer treatment. In addition, this paper provides an in-depth discussion of the future development of this field and proposes a promising method for further research and clinical progress.
ABSTRACT
Transferrin (Tf), widely known for its role as an iron-binding protein, exemplifies multitasking in biological processes. The role of Tf in iron metabolism involves both the uptake of iron from Tf by various cells, as well as the endocytosis mediated by the complex of Tf and the transferrin receptor (TfR). The direct conjugation of the therapeutic compound and immunotoxin studies using Tf peptide or anti-Tf receptor antibodies as targeting moieties aims to prolong drug circulation time and augment efficient cellular drug uptake, diminish systemic toxicity, traverse the blood-brain barrier, restrict systemic exposure, overcome multidrug resistance, and enhance therapeutic efficacy with disease specificity. This review primarily discusses the various biological actions of Tf, as well as the development of Tf-targeted nano-based drug delivery systems. The goal is to establish the use of Tf as a disease-targeting component, accentuating the potential therapeutic applications of this protein.
ABSTRACT
Metabolic dysfunction-associated steatotic liver disease (MASLD) has a global prevalence of 25% and is a leading cause of cirrhosis and hepatocellular carcinoma. The prevalence of MASLD has been increasing, mirroring the global increase in diabetes and metabolic syndrome. MASLD is a chronic and progressive condition characterized by inflammation, oxidative stress, insulin resistance, and disruptions in lipid metabolism. Programmed cell death (PCD) plays a pivotal role in determining the pathological aspects of MASLD, including liver inflammation, fibrosis, and even the potential for malignant transformation. PCD is a dominant process that is fundamental for eukaryotic growth and serves as a regulatory factor in MASLD. PCD encompasses various pathways, including autophagy, ferroptosis, apoptosis, and pyroptosis. These PCD pathways can be activated at different stages of MASLD. The key effector molecules involved in these processes are central focal points in the development of therapeutic interventions for MASLD. Here, we comprehensively review the idea that targeted the modulation of the PCD pathway may be an effective approach for the prevention and/or treatment of MASLD.
Subject(s)
Carcinoma, Hepatocellular , Fatty Liver , Ferroptosis , Liver Neoplasms , Metabolic Diseases , Humans , Pyroptosis , Apoptosis , Autophagy , Carcinoma, Hepatocellular/genetics , InflammationABSTRACT
Hepatic steatosis, which is triggered by dysregulation of lipid metabolism and redox equilibrium in the liver, is regarded as a risk factor in the non-alcoholic fatty liver disease (NAFLD). However, pharmacologic engagement of this process is difficult. We identified the small molecule NSC48160 as an effective agent against nonalcoholic steatohepatitis (NASH). We found that NSC48160 significantly lowered hepatic lipid levels in vitro and in vivo by activating the AMPKα-dependent pathway. AMPKα regulated its downstream pathway involved in lipogenesis (SREBP-1c/FASN pathway) and fatty acid oxidation (PPARα pathway). Metabonomics analysis combined with RNA-sequencing profiling revealed that NSC48160-induced lipogenesis is modulated by lipid metabolism. Moreover, NSC48160 dramatically reduces reactive oxygen species (ROS) production, restores the levels of the membrane potential and NAD+/NADH ratio, and improves mitochondrial respiration. These findings suggest that NSC48160 is a promising hit compound in the pursuit of a pharmacological approach in the treatment of NASH.
ABSTRACT
Immunotherapy is a therapeutic approach that employs immunological principles and techniques to enhance and amplify the body's immune response, thereby eradicating tumor cells. Immunotherapy has demonstrated effective antitumor effects on a variety of malignant tumors. However, when applied to humans, many immunotherapy drugs fail to target lesions with precision, leading to an array of adverse immune-related reactions that profoundly limit the clinical application of immunotherapy. Nanodrug delivery systems enable the precise delivery of immunotherapeutic drugs to targeted tissues or specific immune cells, enhancing the immune antitumor effect while reducing the number of adverse reactions. A nanodrug delivery system provides a feasible strategy for activating the antitumor immune response by the following mechanisms: 1) increased targeting and uptake of vaccines by DCs, which enhances the efficacy of the immune response; 2) increased tumor cell immunogenicity; 3) regulation of TAMs and other cells by, for example, regulating the polarization of TAMs and interfering with TAN formation, and ECM remodeling by CAFs; and 4) interference with tumor immune escape signaling pathways, namely, the PD-1/PD-L1, FGL1/LAG-3 and IDO signaling pathways. This paper reviews the progress of nanodrug delivery system research with respect to tumor immunotherapy based on tumor immunomodulation over the last few years, discussing the promising future of these delivery systems under this domain.
Subject(s)
Nanoparticles , Neoplasms , Humans , Immunomodulation , Immunotherapy , Neoplasms/drug therapy , Immunity , Fibrinogen/pharmacologyABSTRACT
BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is becoming a global epidemic without effective treatment currently available. NAFLD is characterized by an increase in hepatic de novo lipogenesis (DNL) and inadequate compensatory enhancement in fatty acid oxidation (FAO), which disturbs lipid homeostasis. In NAFLD, lipid metabolism relies heavily on metabolic reprogramming. Moreover, lipid metabolism plays an essential role in switching between lipogenesis and FAO, which is beneficial for the anti-NAFLD therapy. Our recent study demonstrated that the phytochemical tetrahydropalmatine (THP) has positive efficacy in hepatocellular carcinoma (HCC). However, it remains unclear whether the therapeutic benefits of THP are primarily due to delaying the progression of hepatic steatosis to HCC. PURPOSE: This work aimed to systemically evaluate the pharmacological functions and underlying mechanisms of THP in NAFLD using both in vitro and in vivo models. METHODS: NAFLD models were established using high-fat diet (HFD)-fed mice in vivo and palmitic acid- and oleic acid-challenged hepatocytes in vitro. Metabonomics analysis concomitant with biochemical indices and computational biology assays were performed comprehensively to reveal the key link between the treatment of NAFLD and the AMPK-SREBP-1c-Sirt1 signaling axis. RESULTS: Hepatic metabolomics analysis revealed that THP altered lipid metabolism by enhancing FAO and inhibiting glycolysis, tricarboxylic acid cycle, and urea cycle in HFD-fed mice. Analysis of gene expression showed that THP profoundly suppressed hepatic DNL and promoted FAO. THP supplementation not only significantly decreased body/liver weight gain and serum indices but also ameliorated hepatic steatosis. Simultaneously, impaired lipotoxicity was observed in vivo and in vitro after THP supplementation, protecting against steatosis-driven injury. Metabolic phenotype assays showed that THP promoted switching from glycolysis inhibition to FAO enhancement in steatotic cells, resulting in reprogramming lipid metabolism. Mechanistically, THP accelerated lipid oxidation by activating AMPK-SREBP-1c-Sirt1 axis signaling. Applying molecular docking combined with surface plasmon resonance and cellular thermal shift assay target engagement, as well as siRNA assays, AMPKα was confirmed as a direct molecular target of THP. CONCLUSION: In summary, THP ameliorates hepatic steatosis in NAFLD by switching lipid metabolism via the AMPK-SREBP-1c-Sirt1 pathway. This work provides an attractive phytochemical component for therapy against hepatic steatosis in NAFLD.
Subject(s)
Carcinoma, Hepatocellular , Liver Neoplasms , Non-alcoholic Fatty Liver Disease , Animals , Mice , Non-alcoholic Fatty Liver Disease/metabolism , Lipid Metabolism , Sterol Regulatory Element Binding Protein 1/metabolism , AMP-Activated Protein Kinases/metabolism , Carcinoma, Hepatocellular/pathology , Sirtuin 1/metabolism , Molecular Docking Simulation , Liver Neoplasms/pathology , Liver , Oleic Acid , Diet, High-Fat/adverse effects , Mice, Inbred C57BLABSTRACT
The regulation of mammalian early-embryonic development is a complex, coordinated process that involves widespread transcriptomic and epigenetic remodeling. The main cause of developmental failure in preimplantation embryos after in vitro fertilization is the irreversible arrested-at-cleavage stage. To deepen our understanding of this embryonic block, we profiled a single-cell multi-omics map of copy number variations (CNVs), the transcriptome, the DNA methylome, and the chromatin state of bovine eight-cell embryos with a two-cell fate that either arrested or developed into blastocysts. To do this, we sequenced a biopsied blastomere and tracked the developmental potential of the remaining cells. Aneuploid embryos inferred by CNVs from DNA- and RNA-library data tended to lose their developmental potency. Analysis of distinct genomic regions of DNA methylation and chromatin accessibility revealed that enrichment of gene function and signaling pathways, such as the MAPK signaling pathway, was altered in arrested euploid eight-cell embryos compared with blastocyst-developed euploid eight-cell embryos. Moreover, the RNA expression and chromatin accessibility of embryonic genome activation-associated genes were lower in arrested euploid embryos than in blastocyst-developed embryos. Taken together, our results indicate that the developmental block of eight-cell embryos can be caused by multiple molecular layers, including CNVs, abnormality of DNA methylation and chromatin accessibility, and insufficient expression of embryonic genome activation-associated genes. Our integrated and comprehensive data set provides a valuable resource to further dissect the exact mechanisms underlying the arrest of bovine eight-cell embryos in vitro.
Subject(s)
DNA Copy Number Variations , Multiomics , Pregnancy , Female , Cattle , Animals , Blastocyst/metabolism , Embryonic Development/genetics , Fertilization in Vitro/veterinary , Chromatin/metabolism , RNA/metabolism , Mammals/geneticsABSTRACT
Non-alcoholic fatty liver disease (NAFLD) is the most prevalent chronic liver disease that affects approximately one-quarter of the global adult population, posing a significant threat to human health with wide-ranging social and economic implications. The main characteristic of NAFLD is considered that the excessive fat is accumulated and deposited in hepatocytes without excess alcohol intake or some other pathological causes. NAFLD is a progressive disease, ranging from steatosis to non-alcoholic steatohepatitis (NASH), cirrhosis, hepatocellular carcinoma, liver transplantation, and death. Therefore, NAFLD will probably emerge as the leading cause of end-stage liver disease in the coming decades. Unlike other highly prevalent diseases, NAFLD has received little attention from the global public health community. Liver biopsy is currently considered the gold standard for the diagnosis and staging of NAFLD because of the absence of noninvasive and specific biomarkers. Due to the complex pathophysiological mechanisms of NAFLD and the heterogeneity of the disease phenotype, no specific pharmacological therapies have been approved for NAFLD at present, although several drugs are in advanced stages of development. This review summarizes the current evidence on the pathogenesis, diagnosis and treatment of NAFLD.
Subject(s)
Carcinoma, Hepatocellular , End Stage Liver Disease , Liver Neoplasms , Non-alcoholic Fatty Liver Disease , Adult , Humans , Non-alcoholic Fatty Liver Disease/diagnosis , Non-alcoholic Fatty Liver Disease/therapy , Non-alcoholic Fatty Liver Disease/epidemiology , Liver Cirrhosis/pathology , Carcinoma, Hepatocellular/pathology , End Stage Liver Disease/pathology , Liver Neoplasms/pathology , Liver/pathologyABSTRACT
OBJECTIVE: Oncolytic adenoviruses are capable of exerting anticancer effects via a variety of mechanisms, including apoptosis and autophagy. In the present study, the dual-specific antitumor oncolytic adenovirus, Ad-Apoptin-hTERT-E1a (ATV), was used to infect cervical cancer cell lines to test its antitumor effects. METHODS: To explore the use of apoptin in tumor gene therapy, a recombinant adenovirus ATV expressing the apoptin protein was assessed to determine its lethal and growth-inhibitory effects on human cervical cancer cell line (HeLa) cells in vitro . Nonapoptotic autophagy of HeLa cells infected with ATV was assessed by examining the cell morphology, development of acidic vesicular organelles and the conversion of microtubule-associated protein 1 light chain 3 (LC3) from its cytoplasmic to autophagosomal membrane form. Using gene silencing (knockdown of LC3 and Belin-1), autophagy-associated molecules (e.g. ATG5, ATG12 and ULK1) were monitored by real-time PCR and western blot. RESULTS: A series of experiments demonstrated that ATV could significantly induce apoptosis and autophagy in cervical cancer cells, and provided evidence that ATV not only induced apoptosis but also autophagy and ATG5, ATG12 and ULK1 related pathways were not entirely dependent on LC3 and Beclin-1. CONCLUSION: These results indicate that ATV may have a potential application in tumor gene therapy.
Subject(s)
Autophagic Cell Death , Oncolytic Virotherapy , Uterine Cervical Neoplasms , Female , Humans , Adenoviridae/genetics , HeLa Cells , Cell Line, Tumor , Apoptosis , Autophagy , Oncolytic Virotherapy/methodsABSTRACT
Nonalcoholic fatty liver disease (NAFLD) constitutes a commonly diagnosed liver pathology with perturbed lipid metabolism, which is mainly caused by excessive accumulation of fat in hepatocytes by various pathogenic factors. Currently, there are no effective drug treatments for NAFLD. Ferroptosis represents a novel form of programmed cell death depending on iron, which is driven by large cellular amounts of reactive oxygen species (ROS) and lipid peroxides. Ferroptosis plays critical regulatory roles in the pathogenesis of NAFLD, and overaccumulation of Fe2+ contributes to lipid peroxidation, which subsequently aggravates NAFLD. Therefore, ferroptosis suppression might constitute an important target for NAFLD treatment. This article reviews the discovery, production pathways, and defense mechanisms of ferroptosis, and explores its association with NAFLD. This may provide new reference targets and strategies for the development of NAFLD drugs from the perspective of ferroptosis.
Subject(s)
Ferroptosis , Non-alcoholic Fatty Liver Disease , Humans , Lipid Peroxidation , Lipid PeroxidesABSTRACT
Pancreatic cancer (PC) is a devastating malignant tumor of gastrointestinal (GI) tumors characterized by late diagnosis, low treatment success and poor prognosis. The most common pathological type of PC is pancreatic ductal adenocarcinoma (PDAC), which accounts for approximately 95% of PC. PDAC is primarily driven by the Kirsten rat sarcoma virus (KRAS) oncogene. Ferroptosis was originally described as ras-dependent cell death but is now defined as a regulated cell death caused by iron accumulation and lipid peroxidation. Recent studies have revealed that ferroptosis plays an important role in the development and therapeutic response of tumors, especially PDAC. As the non-apoptotic cell death, ferroptosis may minimize the emergence of drug resistance for clinical trials of PDAC. This article reviews what has been learned in recent years about the mechanisms of ferroptosis in PDAC, introduces the association between ferroptosis and the KRAS target, and summarizes several potential strategies that are capable of triggering ferroptosis to suppress PDAC progression.
Subject(s)
Carcinoma, Pancreatic Ductal , Ferroptosis , Pancreatic Neoplasms , Humans , Carcinoma, Pancreatic Ductal/drug therapy , Carcinoma, Pancreatic Ductal/genetics , Carcinoma, Pancreatic Ductal/metabolism , Pancreatic Neoplasms/drug therapy , Pancreatic Neoplasms/genetics , Pancreatic Neoplasms/metabolism , Pancreatic Ducts/metabolism , Proto-Oncogene Proteins p21(ras)/genetics , Proto-Oncogene Proteins p21(ras)/metabolism , Pancreatic NeoplasmsABSTRACT
Non-alcoholic fatty liver disease (NAFLD) is the most prevalent chronic liver disease, affecting approximately one-quarter of the global population, and has become a world public health issue. NAFLD is a clinicopathological syndrome characterized by hepatic steatosis, excluding ethanol and other definite liver damage factors. Recent studies have shown that the development of NAFLD is associated with lipid accumulation, oxidative stress, endoplasmic reticulum stress, and lipotoxicity. A range of natural products have been reported as regulators of NAFLD in vivo and in vitro. This paper reviews the pathogenesis of NAFLD and some natural products that have been shown to have therapeutic effects on NAFLD. Our work shows that natural products can be a potential therapeutic option for NAFLD.
Subject(s)
Biological Products , Non-alcoholic Fatty Liver Disease , Humans , Non-alcoholic Fatty Liver Disease/drug therapy , Non-alcoholic Fatty Liver Disease/etiology , Non-alcoholic Fatty Liver Disease/prevention & control , Biological Products/pharmacology , Biological Products/therapeutic use , Biological Products/metabolism , Endoplasmic Reticulum Stress , Oxidative Stress , Ethanol/metabolism , Liver/metabolismABSTRACT
Metabolic dysregulation is a hallmark of hepatocellular carcinoma (HCC). AMPK is a crucial hub of metabolic regulation during cancer progression. We show that phytochemical Levo-tetrahydropalmatine (THP) activates AMPK-dependent autophagy to downregulate the mitochondrial respiration and glycolysis. Consequently, THP significantly decreased cell viability in two HCC cell lines, BEL-7402 and SMMC-7721. Similarly, NOX4 inhibitor diphenyleneiodonium chloride (DPI) induces concomitant downregulation of the mitochondrial and glycolytic metabolism. In contrast to THP, cells are less sensitive to proliferation inhibition induced by DPI treatment as compared to THP treatment did. Combined treatment of THP and DPI was found to be more efficacious in killing cancer cells than either of the agents treated individually. Indeed, the co-operative effect by the THP-DPI combination improves the pro-apoptotic activity in response to the energy depletion as outlined by a drastic decrease in ATP levels. Therapeutic regime significantly reduced the tumor growth in mice. Importantly, this is realized without causing systemic toxicity to other organs. Collectively, our work shows that the combinatorial therapy of autophagy activator THP and NOX4 inhibitor DPI may be considered as a therapeutic avenue against HCC.
Subject(s)
Carcinoma, Hepatocellular , Liver Neoplasms , AMP-Activated Protein Kinases , Animals , Berberine Alkaloids , Carcinoma, Hepatocellular/drug therapy , Liver Neoplasms/drug therapy , Mice , Onium CompoundsABSTRACT
Mitochondrion is an important organelle that maintains cellular homeostasis and plays a crucial role in determining cell fate. The present study investigated the effect of levo-tetrahydropalmatine(THP) on autophagic flux and energy metabolism phenotype of human hepatocellular carcinoma(HCC) SMMC-7721 and BEL-7402 cells. SMMC-7721 and BEL-7402 cells were treated with THP(100 µmol·L~(-1)) with or without N-acetyl-L-cysteine(NAC, 10 µmol·L~(-1)) for 24 h. The mitochondrial reactive oxygen species(mtROS) was detected by flow cytometry(FCM) with MitoSOX probe and fluorescence microscopy, respectively. Thereafter, autophagic flux was detected by FCM with CYTO-ID probe, and the protein levels of microtubule-associated protein 1 A/1 B-light chain 3-â (LC3â ), LC3â ¡, and phosphorylated AMP-activated protein kinase(p-AMPK)/AMPK were measured by Western blot. Mitochondrial respiration was examined by Seahorse XFp assay and cell proliferation by a system. Annexin V-FITC and PI/RNase staining was employed to detect apoptosis of SMMC-7721 and BEL-7402 cells treated with THP and/or NAC. Subsequently, membrane potential was measured with MitoTracker Red CMXRos. Compared with the control group, THP promoted mtROS production and THP combined with NAC attenuated the autophagic flux increase induced by THP alone in SMMC-7721 and BEL-7402 cells. When cells were co-treated with THP and chloroquine(CQ, an autophagy inhibitor), THP further increased mtROS and apoptosis. In addition, THP significantly reduced mitochondrial respiration in terms of mitochondrial basal respiration, ATP production, and maximal respiration. Meanwhile, THP significantly reduced the proliferation index and mitochondrial membrane potential of HCC cells accompanied by the increased apoptosis. This study demonstrates that the up-regulation of mtROS by THP significantly promotes HCC cell autophagy(protective autophagy) and impairs mitochondrial respiration through reprogramming energy metabolism, ultimately inducing the mitochondria-mediated apoptosis of SMMC-7721 and BEL-7402 cells.
Subject(s)
Carcinoma, Hepatocellular , Liver Neoplasms , Humans , Carcinoma, Hepatocellular/drug therapy , Carcinoma, Hepatocellular/genetics , Reactive Oxygen Species/metabolism , Up-Regulation , Liver Neoplasms/drug therapy , Liver Neoplasms/genetics , AMP-Activated Protein Kinases/genetics , AMP-Activated Protein Kinases/metabolism , Mitochondria , Autophagy , Cell Line, Tumor , Apoptosis , PhenotypeABSTRACT
Targeting cancer cell metabolism has been an attractive approach for cancer treatment. However, the role of metabolic alternation in cancer is still unknown whether it functions as a tumor promoter or suppressor. Applying the cancer gene-metabolism integrative network model, we predict adenosine monophosphate-activated protein kinase (AMPK) to function as a central hub of metabolic landscape switching in specific liver cancer subtypes. For the first time, we demonstrate that the phytochemical levo-tetrahydropalmatine (l-THP), a Corydalis yanhusuo-derived clinical drug, as an AMPK activator via autophagy-mediated metabolic switching could kill the hepatocellular carcinoma HepG2 cells. Mechanistically, l-THP promotes the autophagic response by activating the AMPK-mTOR-ULK1 and the ROS-JNK-ATG cascades and impairing the ERK/AKT signaling. All these processes ultimately synergize to induce the decreased mitochondrial oxidative phosphorylation (OXPHOS) and mitochondrial damage. Notably, silencing AMPK significantly inhibits the autophagic flux and recovers the decreased OXPHOS metabolism, which results in HepG2 resistance to l-THP treatment. More importantly, l-THP potently reduces the growth of xenograft HepG2 tumor in nude mice without affecting other organs. From this perspective, our findings support the conclusion that metabolic change is an alternative approach to influence the development of HCC.
ABSTRACT
HRas-GTP has a transient intermediate state with a "non-signaling open conformation" in GTP hydrolysis and nucleotide exchange. Due to the same hydrolysis process and the structural homology, it can be speculated that the active KRas adopts the same characteristics with the "open conformation." This implies that agents locking this "open conformation" may theoretically block KRas-dependent signaling. Applying our specificity-affinity drug screening approach, NSC290956 was chosen by high affinity and specificity interaction with the "open conformation" structure HRasG60A-GppNp. In mutant KRas-driven non-small-cell lung cancer (NSCLC) model system, NSC290956 effectively suppresses the KRas-GTP state and gives pharmacological KRas inhibition with concomitant blockages of both the MAPK-ERK and AKT-mTOR pathways. The dual inhibitory effects lead to the metabolic phenotype switching from glycolysis to mitochondrial metabolism, which promotes the cancer cell death. In the xenograft model, NSC290956 significantly reduces H358 tumor growth in nude mice by mechanisms similar to those observed in the cells. Our work indicates that NSC290956 can be a promising agent for the mutant KRas-driven NSCLC therapy.
ABSTRACT
Autophagy and apoptosis both promote cell death; however, the relationship between them is subtle, and they mutually promote and antagonize each other. Apoptin can induce apoptosis of various tumor cells; however, tumor cell death is not only caused by apoptosis. Whether apoptin affects tumor cell autophagy is poorly understood. Therefore, the present study aimed to explore the potential mechanisms underlying the effects of apoptin using recombinant adenoviruses expressing apoptin. Reverse transcriptionquantitative polymerase chain reaction, immunoblotting, flow cytometry, fluorescence microscopy and proteomics analyses revealed that apoptin could induce autophagy in MCF7 breast cancer cells. The results also suggested that apoptin affected autophagy in a time and dosedependent manner. During the early stage of apoptin stimulation (6 and 12 h), the expression levels of autophagy pathwayassociated proteins, including Beclin1, microtubuleassociated protein 1A/1Blight chain 3, autophagyrelated 4B cysteine peptidase and autophagyrelated 5, were significantly increased, suggesting that apoptin promoted the upregulation of autophagy in MCF7 cells. Conversely, after 12 h of apoptin stimulation, the expression levels of apoptosisassociated proteins were decreased, thus suggesting that apoptosis may be inhibited. Therefore, it was hypothesized that apoptin may enhance autophagy and inhibit apoptosis in MCF7 cells at the early stage. In conclusion, apoptininduced cell death may involve both autophagy and apoptosis. The induction of autophagy may inhibit apoptosis, whereas apoptosis may inhibit autophagy; however, occasionally both pathways operate at the same time and involve apoptin. This apoptinassociated selection between tumor cell survival and death may provide a potential therapeutic strategy for breast cancer.
Subject(s)
Autophagy/genetics , Breast Neoplasms/therapy , Capsid Proteins/genetics , Oncolytic Virotherapy/methods , Adenoviridae/genetics , Apoptosis/drug effects , Breast Neoplasms/pathology , Cell Proliferation/drug effects , Cell Survival/drug effects , Drug Screening Assays, Antitumor , Female , Genetic Vectors/genetics , Humans , MCF-7 Cells , Oncolytic Viruses/geneticsABSTRACT
rVTT-TA35-TJ, an attenuated vaccinia virus Tian Tan strain (VTT), was constructed by knocking out two non-essential gene fragments (TA35R and TJ2R) related to virulence, immunomodulation, and host range; and by combining double marker screening with exogenous and endogenous selectable marker knock-out techniques. Here, the shuttle plasmids pSK-TA35 and pSK-TJ were constructed, containing two pairs of recombinant arms: early and late strong promoter pE/L and EGFP as an exogenous selectable marker. The recombinant vaccinia virus rVTT-TA35-TJ without exogenous selection markers was then obtained through homologous recombination technology and the Cre/loxP system. Knocking out the two gene fragments does not affect the replication ability of the virus and displays a good genetic stability. Furthermore, a series of in vivo and in vitro experiments demonstrate that although virulence of rVTT-TA35-TJ is attenuated significantly, high immunogenicity was maintained. These results support the potential development of rVTT-TA35-TJ as a safe viral vector or vaccine.
Subject(s)
Drug Carriers , Gene Knockout Techniques , Smallpox Vaccine/genetics , Smallpox Vaccine/immunology , Vaccinia virus/genetics , Vaccinia virus/immunology , Animals , Cell Line , Humans , Mice, Inbred BALB C , Rabbits , Vaccines, Attenuated/genetics , Vaccines, Attenuated/immunology , Vaccinia virus/pathogenicity , Virulence , Virulence Factors/geneticsABSTRACT
Goatpox virus (GTPV) is prevalent in goats and is associated with high mortality. This virus causes fever, skin nodules, lesions in the respiratory and lymph node enlargement. Considering the safety risks and side effects of vaccination with attenuated live GPTV vaccine strain AV41, an attenuated goatpox virus (GTPV-TK-ORF), was constructed by deleting non-essential gene fragments without affecting replication and related to the virulence and immunomodulatory functions of the goatpox virus AV41 strain (GTPV-AV41) using homologous recombination and the Cre (Cyclization Recombination Enzyme)/Loxp system. The results of both in vivo and in vitro experiments demonstrated that GTPV-TK-ORF was safer than wild type GTPV-AV41, possessed satisfactory immunogenicity, and could protect goats from a virulent GTPV-AV40 infection. Moreover, the IFN-γ, GTPV-specific antibody, and neutralizing antibody levels in the GTPV-TK-ORF-immunized group were significantly higher than that in the normal saline control group following immunization (Pâ¯<â¯0.01). Thus, GTPV-TK-ORF may be used as a potential novel vaccine and viral vector with good safety and immunogenicity.
Subject(s)
Capripoxvirus/growth & development , Capripoxvirus/genetics , Gene Deletion , Goat Diseases/prevention & control , Poxviridae Infections/veterinary , Viral Proteins/genetics , Viral Vaccines/administration & dosage , Animals , Antibodies, Neutralizing/blood , Antibodies, Viral/blood , Capripoxvirus/pathogenicity , Cell Line , Gene Knockout Techniques , Goats , Poxviridae Infections/prevention & control , Treatment Outcome , Vaccines, Attenuated/administration & dosage , Vaccines, Attenuated/adverse effects , Vaccines, Attenuated/genetics , Vaccines, Attenuated/isolation & purification , Viral Vaccines/adverse effects , Viral Vaccines/genetics , Viral Vaccines/isolation & purification , VirulenceABSTRACT
Newcastle disease virus (NDV) can specifically kill cancer cells and has less toxicity to normal cells. The hemagglutinin-neuraminidase (HN) protein is an important structural protein in NDV pathogenesis and has been postulated as a promising candidate for antitumor therapy. The aim of this study was to investigate the anticancer potential of recombinant adenovirus Ad-HN-PEG3p-E1a. An MTS assay was performed to determine viral proliferation after viral infection, the data showed that the proliferation ability of osteosarcoma cells decreased, whereas there was no significant change in normal hepatic cells. DAPI and Annexin V experiments showed that osteosarcoma cells were killed because of apoptosis, active oxygen content, and augmented mitochondrial membrane potential loss. Caspase Activity Assay Kits were used to detect the caspase-3 activities of the treated OS-732 for increased expression. Western blot analysis showed that cytochrome C increased significantly and apoptosis of the virus was confirmed in tumor cells. In-vivo experiments show that NDV has an inhibitory effect on tumor growth. The recombinant adenovirus, which is composed of a HN protein and progressive increment promoter PEG3p, could inhibit the growth of OS-732 and promote the apoptosis of tumor cells. However, there was no clear relationship with normal cell (L02) apoptosis.