scholarly journals A Facilitated Diffusion Mechanism Establishes the Drosophila Dorsal Gradient

2016 ◽  
Author(s):  
Sophia N. Carrell ◽  
Michael D. O’Connell ◽  
Amy E. Allen ◽  
Stephanie M. Smith ◽  
Gregory T. Reeves
Keyword(s):  
Target Genes ◽  
Diffusion Mechanism ◽  
Ventral Side ◽  
Inflammatory Factor ◽  
Facilitated Diffusion ◽  
Dependent Manner ◽  
Predominant Role ◽  
Concentration Dependent Manner ◽  
Ventral Midline ◽  
Toll Signaling

SummaryThe transcription factor NF-κB plays an important role in the immune system as an apoptotic and inflammatory factor. In the Drosophila melanogaster embryo, a homolog of NF-ΚB called Dorsal (dl) patterns the dorsal-ventral (DV) axis in a concentration-dependent manner. During early development, dl is sequestered outside the nucleus by Cactus (Cact), homologous to IκB. Toll signaling at the ventral midline breaks the dl/Cact complex, allowing dl to enter the nucleus where it transcribes target genes. Here we show that dl accumulates on the ventral side of the embryo over the last 5 cleavage cycles and that this accumulation is the result of facilitated diffusion of dl/Cact complex. We speculate that the predominant role for Cact in DV axis specification is to shuttle dl towards the ventral midline. Given that this mechanism has been found in other, independent systems, we suggest it may be more prevalent than previously thought.

Anti-tumor activity of SKLB-0322, a novel EZH2 covalent inhibitor, in ovarian cancer.

2021 ◽  
Vol 39 (15_suppl) ◽  
pp. e15052-e15052
Author(s):  
Yongxia Zhu ◽  
Xinyi Chen ◽  
Qiangsheng Zhang ◽  
Lihong Shi ◽  
Luoting Yu ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Target Genes ◽  
Competitive Inhibitor ◽  
Negative Control ◽  
Phase Cell ◽  
Dependent Manner ◽  
Independent Manner ◽  
Western Blot Assay ◽  
Concentration Dependent Manner ◽  
Covalent Inhibitor

e15052 Background: Enhancer of Zeste Homolog 2 (EZH2) is the catalytic subunit of the Polycomb Repressive Complex 2 (PRC2) that regulate downstream target genes expression, and then promotes tumor cell proliferation, metastasis and drug resistance. EZH2 also performs some functions in a PRC2-independent manner. Most of reported EZH2 inhibitors are S-adenosyle-methionine (SAM)-competitive inhibitor, and are less selective for EZH2 close homolog EZH1, which resulted in safety concerns and insufficient efficacy. To obtain irreversible EZH2 inhibitor, a novel covalent inhibitor was developed and characterized. Methods: SKLB-0322 and its derivatives were designed, synthesized and confirmed as EZH2 covalent inhibitor by us. The anti-tumor activities of SKLB-0322 were investigated by MTT assay, flow cytometry, and western blot assay. The reversible analog of SKLB-0322 (SKLB-0322’) was used as negative control. Results: SKLB-0322 inhibited EZH2 methyltransferase activity with nanomolar potency, while the inhibitory activities of SKLB-0322’ was reduced. The mass spectrometry (MS) analyses revealed that SKLB-0322 could efficiently forms a single modified covalent adduct. SKLB-0322 displayed noteworthy potency against ovarian cancer cell lines at low micromolar level and reduced the expression level of H3K27me3 in a concentration-dependent manner, which was about 5-fold more active than the reversible negative control SKLB-0322’. Besides, SKLB-0322 caused G2/M phase cell cycle arrest in A2780 and PA-1 cells. Furthermore, SKLB-0322 induced A2780 and PA-1 cell apoptosis in a time- and concentration- dependent manner. Conclusions: Our data clarified that SKLB-0322 is an EZH2 covalent inhibitor for ovarian cancer therapy which is worthy of further evaluation.

scholarly journals Coffee polyphenols modulate whole-body substrate oxidation and suppress postprandial hyperglycaemia, hyperinsulinaemia and hyperlipidaemia

2011 ◽  
Vol 107 (12) ◽  
pp. 1757-1765 ◽  
Author(s):  
Takatoshi Murase ◽  
Yuka Yokoi ◽  
Koichi Misawa ◽  
Hideo Ominami ◽  
Yasuto Suzuki ◽  
...  
Keyword(s):  
Energy Metabolism ◽  
Digestive Enzymes ◽  
Substrate Oxidation ◽  
Postprandial Hyperglycaemia ◽  
Whole Body ◽  
Dependent Manner ◽  
Predominant Role ◽  
Concentration Dependent Manner ◽  
Potent Inhibitory Activity

Postprandial energy metabolism, including postprandial hyperglycaemia, hyperinsulinaemia and hyperlipidaemia, is related to the risk for developing obesity and CVD. In the present study, we examined the effects of polyphenols purified from coffee (coffee polyphenols (CPP)) on postprandial carbohydrate and lipid metabolism, and whole-body substrate oxidation in C57BL/6J mice. In mice that co-ingested CPP with a lipid–carbohydrate (sucrose or starch)-mixed emulsion, the respiratory quotient determined by indirect calorimetry was significantly lower than that in control mice, whereas there was no difference inVO2(energy expenditure), indicating that CPP modulates postprandial energy partitioning. CPP also suppressed postprandial increases in plasma glucose, insulin, glucose-dependent insulinotropic polypeptide and TAG levels. Inhibition experiments on digestive enzymes revealed that CPP inhibits maltase and sucrase, and, to a lesser extent, pancreatic lipase in a concentration-dependent manner. Among the nine kinds of polyphenols (caffeoyl quinic acids (CQA), di-CQA, feruloyl quinic acids (FQA)) contained in CPP, di-CQA showed more potent inhibitory activity than CQA or FQA on these digestive enzymes, suggesting a predominant role of di-CQA in the regulation of postprandial energy metabolism. These results suggest that CPP modulates whole-body substrate oxidation by suppressing postprandial hyperglycaemia and hyperinsulinaemia, and these effects are mediated by inhibiting digestive enzymes.

scholarly journals Aspirin inhibits tumor progression and enhances cisplatin sensitivity in epithelial ovarian cancer

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11591
Author(s):  
Jianfeng Guo ◽  
Yapei Zhu ◽  
Lili Yu ◽  
Yuan Li ◽  
Jing Guo ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Tumor Growth ◽  
Growth Inhibition ◽  
Epithelial Ovarian Cancer ◽  
Target Genes ◽  
Tumor Growth Inhibition ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
P53 Target Genes

Background Ovarian cancer is the most common gynecological malignancy and is difficult to manage due to the emergence of resistance to various chemotherapeutic drugs. New efforts are urgently awaited. Aspirin, which is traditionally considered a nonsteroidal anti-inflammatory drug (NSAID), has been reported to exert potential chemopreventive effects. Therefore, we aimed to investigate the anticancer effect and explore the underlying molecular mechanisms of aspirin on epithelial ovarian cancer (EOC) cells. Methods We conducted wound healing, transwell migration, EdU cell proliferation, colony formation and apoptosis detection assays to observe the effects of aspirin on the migration, proliferation and apoptosis of EOC cells (A2870, Caov-3, and SK-OV-3). EOC cells were treated with a combination of aspirin and cisplatin (CDDP) to observe the effect of aspirin on enhancing CDDP sensitivity. Orthotopic xenograft models of ovarian cancer established with A2780-Luciferase-GFP cells were applied to compare tumor growth inhibition in the control, CDDP and CDDP plus aspirin groups through in vivo imaging, which can be used to continuously monitor tumor growth. The expression and acetylation levels of p53 in EOC cells treated with aspirin were determined using western blotting, and p53 acetylation levels were examined in tumors harvested from the transplanted mice. Quantitative real-time PCR was used to assess the mRNA expression of p53 target genes. Results Aspirin inhibited migration and proliferation and induced apoptosis in EOC cell lines in a concentration-dependent manner. In vitro, aspirin enhanced the sensitivity of EOC cells to CDDP by increasing its inhibitory effect on proliferation and its effect on inducing apoptosis. In vivo, the differences in the tumor growth inhibition rates among the different CDDP experimental groups were statistically significant (p < 0.05). Aspirin did not affect p53 protein expression but increased the p53 acetylation level in a concentration-dependent manner. In addition, the mRNA levels of CDKN1A, BAX, FOXF1, PUMA, and RRAD in EOC cells were significantly increased by the aspirin treatment. Conclusions Aspirin inhibits tumor progression and enhances the CDDP sensitivity of EOC cells. These antitumor effects of aspirin might be mediated by p53 acetylation and subsequent activation of p53 target genes.

scholarly journals Global transcriptome analysis reveals partial estrogen-like effects of karanjin in MCF-7 breast cancer cells

2021 ◽  
Author(s):  
Gaurav Bhatt ◽  
Akshita Gupta ◽  
Latha Rangan ◽  
Anil Mukund Limaye
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Target Genes ◽  
Biological Activities ◽  
Biological Effects ◽  
Dependent Manner ◽  
Cell Type ◽  
Concentration Dependent Manner ◽  
Mcf 7

Karanjin, an abundantly occurring furanoflavonoid in edible and non-edible legumes, exerts diverse biological effects in vivo, and in vitro. Its potential as an anticancer agent is also gaining traction following recent demonstrations of its anti-proliferative, cell cycle inhibitory, and pro-apoptotic effects. However, the universality of its anticancer potential is yet to be scrutinized, particularly so because flavonoids can act as selective estrogen receptor modulators (SERMs). Even the genomic correlates of its biological activities are yet to be examined in hormone responsive cells. This paper presents the early and direct transcriptomic footprint of 10 μM karanjin in MCF-7 breast cancer cells, using next generation sequencing technology (RNA-seq). We show that karanjin-modulated gene-expression repertoire is enriched in several hallmark gene sets, which include early estrogen-response, and G2/M checkpoint genes. Genes modulated by karanjin overlapped with those modulated by 1 nM 17β-estradiol (E2), or 1 μM tamoxifen. Karanjin altered the expression of selected estrogen-regulated genes in a cell-type, and concentration dependent manner. It downmodulated the expression of ERα protein in MCF-7 cells. Furthermore, ERα knockdown negatively impacted karanjins ability to modulate the expression of selected E2 target genes. Our data suggest that karanjin exerts its effects on ERα-positive breast cancer cells, at least in part, via ERα. The apparent SERM-like effects of karanjin pose a caveat to the anticancer potential of karanjin. In-depth studies on cell-type and concentration-dependent effects of karanjin may bring out its true potential in endocrine therapies.

Pelle kinase is activated by autophosphorylation during Toll signaling in Drosophila

Development ◽  
2002 ◽  
Vol 129 (8) ◽  
pp. 1925-1933 ◽  
Author(s):  
Baohe Shen ◽  
James L. Manley
Keyword(s):  
Dependent Manner ◽  
Loss Of Function ◽  
Concentration Dependent Manner ◽  
Downstream Target ◽  
Toll Signaling ◽  
Early Embryos ◽  
High Concentrations ◽  
L2 Cells

The Drosophila Pelle kinase plays a key role in the evolutionarily conserved Toll signaling pathway, but the mechanism responsible for its activation has been unknown. We present in vivo and in vitro evidence establishing an important role for concentration-dependent autophosphorylation in the signaling process. We first show that Pelle phosphorylation can be detected transiently in early embryos, concomitant with activation of signaling. Importantly, Pelle phosphorylation is enhanced in a gain-of-function Toll mutant (Toll10b), but decreased by loss-of-function Toll alleles. Next we found that Pelle is phosphorylated in transfected Schneider L2 cells in a concentration-dependent manner such that significant modification is observed only at high Pelle concentrations, which coincide with levels required for phosphorylation and activation of the downstream target, Dorsal. Pelle phosphorylation is also enhanced in L2 cells co-expressing Toll10b, and is dependent on Pelle kinase activity. In vitro kinase assays revealed that recombinant, autophosphorylated Pelle is far more active than unphosphorylated Pelle. Importantly, unphosphorylated Pelle becomes autophosphorylated, and activated, by incubation at high concentrations. We discuss these results in the context of Toll-like receptor mediated signaling in both flies and mammals.

scholarly journals Targeted Delivery of Gene Silencing in Fungi Using Genetically Engineered Bacteria

2021 ◽  
Vol 7 (2) ◽  
pp. 125
Author(s):  
Jonatan Niño-Sánchez ◽  
Li-Hung Chen ◽  
Jorge Teodoro De Souza ◽  
Sandra Mosquera ◽  
Ioannis Stergiopoulos
Keyword(s):  
Targeted Delivery ◽  
Pathogenic Bacteria ◽  
Target Genes ◽  
Genetically Engineered ◽  
Biologically Active ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Whole Cell ◽  
Thaw Cycle ◽  
Living Bacteria

Exploiting RNA interference (RNAi) in disease control through non-transformative methods that overcome the hurdle of producing transgenic plants has attracted much attention over the last years. Here, we explored such a method and used non-pathogenic bacteria as a versatile system for delivering RNAi to fungi. Specifically, the RNaseIII-null mutant strain of Escherichia coli HT115(DE3) was transformed with two plasmid vectors that enabled the constitutive or IPTG-inducible production of double-stranded RNAs (dsRNAs) against genes involved in aflatoxins production in Aspergillus flavus (AflC) or virulence of Botrytis cinerea (BcSAS1). To facilitate the release of the dsRNAs, the bacterial cells were further genetically engineered to undergo a bacteriophage endolysin R-mediated autolysis, following a freeze-thaw cycle. Exposure under in vitro conditions of A. flavus or B. cinerea to living bacteria or their whole-cell autolysates induced silencing of AflC and BcSAS1 in a bacteria concentration-dependent manner, and instigated a reduction in aflatoxins production and mycelial growth, respectively. In planta applications of the living bacteria or their crude whole-cell autolysates produced similar results, thus creating a basis for translational research. These results demonstrate that bacteria can produce biologically active dsRNA against target genes in fungi and that bacteria-mediated RNAi can be used to control fungal pathogens.

scholarly journals N-Phenyl Cinnamamide Derivatives Protect Hepatocytes against Oxidative Stress by Inducing Cellular Glutathione Synthesis via Nuclear Factor (Erythroid-Derived 2)-Like 2 Activation

Molecules ◽  
2021 ◽  
Vol 26 (4) ◽  
pp. 1027
Author(s):  
Sou Hyun Kim ◽  
Minwoo Kim ◽  
Doyoung Kwon ◽  
Jae Sung Pyo ◽  
Joo Hyun Kim ◽  
...  
Keyword(s):  
Target Genes ◽  
Cell Injury ◽  
Antioxidant Response ◽  
Luciferase Reporter ◽  
Dependent Manner ◽  
Nuclear Factor ◽  
Quinone Oxidoreductase ◽  
Concentration Dependent Manner ◽  
Endogenous Antioxidant ◽  
Antioxidative Therapy

Substituted N-phenyl cinnamamide derivatives were designed and synthesized to confirm activation of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) pathway by the electronic effect on beta-position of Michael acceptor according to introducing the R1 and R2 group. Compounds were screened using the Nrf2/antioxidant response element (ARE)-driven luciferase reporter assay. Compound 1g showed desirable luciferase activity in HepG2 cells without cell toxicity. mRNA and protein expression of Nrf2/ARE target genes such as NAD(P)H quinone oxidoreductase 1, hemeoxygenase-1, and glutamate-cysteine ligase catalytic subunit (GCLC) were upregulated by compound 1g in a concentration-dependent manner. Treatment with 1g resulted in increased endogenous antioxidant glutathione, showing strong correlation with enhanced GCLC expression for synthesis of glutathione. In addition, tert-butyl hydroperoxide (t-BHP)-generated reactive oxygen species were significantly removed by 1g, and the results of a cell survival assay in a t-BHP-induced oxidative cell injury model showed a cytoprotective effect of 1g in a concentration dependent manner. In conclusion, the novel compound 1g can be utilized as an Nrf2/ARE activator in antioxidative therapy.

scholarly journals A Novel function for Cactus/IκB inhibitor to promote Toll signals in the Drosophila embryo

2016 ◽  
Author(s):  
Maira Arruda Cardoso ◽  
Marcio Fontenele ◽  
Bomyi Lim ◽  
Paulo Mascarello Bisch ◽  
Stanislav Shvartsman ◽  
...  
Keyword(s):  
Gene Expression ◽  
Innate Immunity ◽  
Nuclear Localization ◽  
Target Genes ◽  
Ventral Side ◽  
Drosophila Embryo ◽  
Embryonic Patterning ◽  
Toll Signaling ◽  
Pathway Activation ◽  
Evolutionarily Conserved

AbstractThe evolutionarily conserved Toll signaling pathway controls innate immunity across phyla and embryonic patterning in insects. In the Drosophila embryo Toll is required to establish gene expression domains along the dorsal-ventral axis. Pathway activation induces degradation of the IκB inhibitor Cactus resulting in a nuclear gradient of the NFκB effector Dorsal. Here we investigate how cactus modulates Toll signals through its effects on the Dorsal gradient and Dorsal target genes. Quantitative analysis using a series of loss and gain-of-function conditions shows that the ventral and lateral aspects of the Dorsal gradient behave differently respective to Cactus fluctuations. Unexpectedly, Cactus favors Dorsal nuclear localization required as response to high Toll signals at the ventral side of the embryo. Furthermore, N-terminal deleted Cactus mimics these effects, indicating that the ability of Cactus to favor Toll stems from mobilization of a free Cactus pool induced by the Calpain A protease. These results indicate that unexplored mechanisms are at play to ensure a correct response to high Toll signals.Summary:The IκB protein Cactus favors high Toll signals, revealing that the ventral and lateral aspects of the Dorsal/NFκB nuclear gradient behave differently respective to Cactus concentrations in the Drosophila embryo.

scholarly journals Troxerutin Stimulates Osteoblast Differentiation of Mesenchymal Stem Cell and Facilitates Bone Fracture Healing

2021 ◽  
Vol 12 ◽  
Author(s):  
Xiao Yang ◽  
Jiang Shao ◽  
Xiao-Min Wu ◽  
Fei-Fei Pan ◽  
Shao-An Yang ◽  
...  
Keyword(s):  
Fracture Healing ◽  
Bone Fracture ◽  
Target Genes ◽  
Fracture Repair ◽  
Marker Genes ◽  
Nodule Formation ◽  
Dependent Manner ◽  
Concentration Dependent Manner

Troxerutin (TRX), a semi-synthetic derivative of the natural bioflavonoid rutin, is a bioactive flavonoid widely abundant in various fruits and vegetables. Known as vitamin P4, TRX has been demonstrated to have several activities including anti-inflammation, anti-oxidants, vasoprotection, and immune support in various studies. Although rutin, the precursor of troxerutin, was reported to have a protective role against bone loss, the function of TRX in skeletal system remains unknown. In the present study, we found that TRX promoted osteogenic differentiation of human mesenchymal stem cells (MSCs) in a concentration-dependent manner by stimulating the alkaline phosphatase (ALP) activity, calcium nodule formation and osteogenic marker genes expression in vitro. The further investigation demonstrated that TRX stimulated the expression of the critical transcription factor β-catenin and several downstream target genes of Wnt signaling, thus activated Wnt/β-catenin signaling. Using a femur fracture rats model, TRX was found to stimulate new bone formation and accelerate the fracture healing in vivo. Collectively, our data demonstrated that TRX could promote osteogenesis in vitro and facilitate the fracture healing in vivo, indicating that TRX may be a promising therapeutic candidate for bone fracture repair.

Long-range action of Wingless organizes the dorsal-ventral axis of the Drosophila wing

Development ◽  
1997 ◽  
Vol 124 (4) ◽  
pp. 871-880 ◽  
Author(s):  
C.J. Neumann ◽  
S.M. Cohen
Keyword(s):  
Cell Fate ◽  
Long Range ◽  
Target Genes ◽  
Dependent Manner ◽  
Short Range Interaction ◽  
Range Interaction ◽  
Concentration Dependent Manner ◽  
Drosophila Wing ◽  
Compartment Boundary ◽  
Organizing Center

Short-range interaction between dorsal and ventral (D and V) cells establishes an organizing center at the DV compartment boundary that controls growth and specifies cell fate along the dorsal-ventral axis of the Drosophila wing. The secreted signaling molecule Wingless (Wg) is expressed by cells at the DV compartment boundary and has been implicated in mediating its long-range patterning activities. Here we show that Wg acts directly, at long range, to define the expression domains of its target genes, Distal-less and vestigial. Expression of the Achaete-scute genes, Distal-less and vestigial at different distances from the DV boundary is controlled by Wg in a concentration-dependent manner. We propose that Wg acts as a morphogen in patterning the D/V axis of the wing.

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