Cycle Progression Research Articles

TEMPORAL VARIATION AND SPATIAL DISTRIBUTION OF RELATIVE INDICES OF LEAF CHLOROPHYLL IN GRAPEVINE cv. CHARDONNAY

Variation in the spatial distribution of leaf chlorophyll content associated with the progression of the phenological cycle of crops may occur in cultivated areas as a result of the variability of environmental conditions and of the intrinsic properties of the plants. The objective of the present study was to model the trend in variation and assess the temporal stability of index of chlorophyll a, b, and total chlorophyll (Chla, Chlb, and Chlt, respectively), and to characterize the spatial distribution of Chlt index in grapevine cv. ‘Chardonnay’. The assessments consisted of in situ measurements made with a portable meter in a commercial vineyard located in the municipality of Espírito Santo do Pinhal, state of São Paulo, Brazil, in the period between flowering and fruit maturation. Descriptive statistics were applied to the indices and regression models were fitted to ascertain the relationship of their mean variation with time. The temporal stability of Chlt index was estimated using Spearman's rank correlation analysis and thematic maps were created using geostatistical analysis and spatial estimation by ordinary kriging. The Chlb and Chlt indices were non-linearly associated with cycle progression and their decrease after the start of maturation was estimated. The temporal stability of the Chlt index was low and variation in its spatial distribution was observed over the assessed period.

Effects of rotary cell culture system‐simulated microgravity on the ultrastructure and biological behavior of human MDA‐MB‐231 breast cancer cells

AbstractObjectiveThis study aimed to investigate changes in the ultrastructure, apoptosis, cycle progression, and migration ability of human MDA‐MB‐231 breast cancer cells under simulated microgravity conditions.MethodsMDA‐MB‐231 breast cancer cells were cultured in a rotary cell culture system bioreactor to simulate microgravity for 7 days. The cells were randomly divided into a simulated microgravity (SMG) group and normal gravity control (NG) group. Changes in apoptosis, ultrastructure, cell cycle, and migration ability were evaluated.ResultsTransmission electron microscopy showed that cells in the SMG group had more secondary lysosomes than the NG groups. The flow cytometry assay showed that the proportion of apoptotic cells in the SMG group was significantly increased compared with the NG group; additionally, the median and interquartile spacing of the proportion of cells in the G0/G1 phase in the SMG group was lower than in the NG group, but the proportion of cells in the S phase was increased in the SMG group compared with the NG group. The transwell migration assay showed that cell migration in the SMG group was significantly reduced compared with the NG group. The expression of BCL‐2 and MMP9 in the SMG group was decreased compared with the NG group, whereas the expression of cyclin D3 in the SMG group was increased.ConclusionSimulated microgravity through rotary cell culture system had significant effects on human MDA‐MB‐231 breast cancer cells: the cell cycle and ultrastructure were impaired, apoptosis increased, migration ability decreased, and the expression of BCL‐2, cyclin D3, and MMP9 were affected.

Circular RNA hsa_circRNA_103809 promoted hepatocellular carcinoma development by regulating miR-377-3p/FGFR1/ERK axis.

In the last decade, circular RNAs (circRNAs) emerge as important regulators in multiple biological processes. Lately, it is reported hsa_circRNA_103809 could play vital parts in several types of cancers. Based on the analysis of GEO data (GSE97332), hsa_circRNA_103809 was found to be dysregulated in hepatocellular carcinoma (HCC). However, the biological function and underlying regulatory mechanisms of hsa_circRNA_103809 in HCC remain unclear. Our results suggested that hsa_circRNA_103809 was overexpressed in HCC patients, and hsa_circRNA_103809 knockdown remarkably inhibited the proliferation, cycle progression, and migration of HCC cells. The investigations of molecular showed that hsa_circRNA_103809 could elevate the protein expression of a miR-377-3p target, fibroblast growth factor receptor 1 (FGFR1), through interacting with miR-377-3p and decreasing its expression level. Additionally, in vivo assays revealed hsa_circRNA_103809 short hairpin RNA served as a tumor suppressor through downregulating FGFR1 in HCC. This study systematically investigated novel regulatory signaling of hsa_circRNA_103809/miR-377-3p/FGFR1 axis, providing insights into hepatocellular carcinoma treatment from bench to clinic.

Characterization of the human E2F4 promoter region and its response to 12-O-tetradecanoylphorbol-13-acetate.

The E2F transcription factors (TFs), which control the progression of the cell cycle in response to DNA-damage and various stresses, are known to interact with a tumor suppressor, Retinoblastoma 1 (RB1). We previously showed that the response of the human RB1 promoter to a 12-O-tetradecanoylphorbol-13-acetate (TPA) in HL-60 cells is mediated by a duplicated GGAA-motif, which is also present in the 5'-upstream of the E2F family genes. The motifs are especially rich in the 5'-upstream of the E2F4 gene. In the present study, we constructed Luciferase (Luc) expression vectors containing a 466-bp of the 5'-upstream of the human E2F4 gene. The transfection of this plasmid and deletion/mutation-introduced derivatives into HL-60 cells and a Luc reporter assay showed that duplicated and triplicated GGAA (TTCC) motifs in the E2F4 promoter respond to TPA. As expected, electrophoretic mobility shift assay (EMSA) indicated that SPI1 (PU.1) binds to the GGAA-motif containing element. A quantitative RT-PCR and Western blotting showed that the E2F4 transcripts and its encoding proteins accumulate during the differentiation of HL-60 into macrophage-like cells. In contrast, the expression of the E2F1 gene and the protein, which possibly acts as a cell cycle accelerator, was greatly diminished.

Long noncoding RNA TUSC7 inhibits cell proliferation, migration and invasion by regulating SOCS4 (SOCS5) expression through targeting miR-616 in endometrial carcinoma

BackgroundLong non-coding RNA (lncRNA) is emerging as an important regulator in various physiological and pathological processes. Recently, it was found that lncRNA long non-coding RNA tumor suppressor candidate 7 (TUSC7) could play tumor suppressive roles in several cancers. However, the function and underlying regulatory mechanism of lncRNA TUSC7 in endometrial carcinoma (EC) remains largely unclear. MethodsThe expression levels of TUSC7 and microRNAs-616 (miR-616) were analyzed by real-time PCR and in situ hybridization. Cell cycle and cell metastasis associated protein expressions were determined by western blotting. Cell proliferation, cycle and metastasis were determined by CCK-8 cell viability, colony formation, flow cytometer, wound scratch and transwell assays respectively in vitro. RNA pull-down, luciferase and western blotting assays were used to examine the target relationship between TUSC7 and miR-616 or that between miR-616 and suppressors of cytokine signaling 4 (5) (SOCS4 (SOCS5)). The functional effects of TUSC7 through sponging miR-616 were further examined using a xenograft tumor mouse model in vivo. ResultsTUSC7 was downexpressed in EC tissues and cell lines, and TUSC7 upregulation could remarkably inhibit cell proliferation, cycle progression and metastasis in EC cells. Mechanistic investigations demonstrated that TUSC7 can interact with miR-616 and decrease its expression, thereby upregulating the expression of miR-616's targets SOCS4 (SOCS5). Additionally, in vivo experiments using a xenograft tumor mouse model revealed that TUSC7 can serve as a tumor suppressor through sponging miR-616, and upregulating SOCS4 (SOCS5) in EC. ConclusionsIn this study, a newly identified regulatory mechanism of lncRNA TUSC7/miR-616/ SOCS4 (SOCS5) axis was systematically studied, which may hold promise as a promising target for EC treatment.

WD repeat and SOCS box containing protein 2 in the proliferation, cycle progression, and migration of melanoma cells

BackgroundWD repeat and SOCS box containing protein (WSB) molecules have important roles in tumorigenesis. WSB1 is dysfunctional in many malignancies. However, the effects of WSB2 in tumors, including melanoma, have not been reported. Here, we investigated the effects of WSB2 in melanoma cell proliferation, cycle progression, and migration, and the underlying mechanisms. MethodsFirst, WSB2 expression levels and their association with clinicopathological features were evaluated in human melanoma tissue samples. Then, WSB2 was knocked down, using specific shRNA, in melanoma A375 and G361 cells. Proliferation, cycle progression, and migration of A375 and G361 cells were evaluated by 3-(4,5-diethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-etrazolium, inner salt (MTS), colony formation, 5-ethynyl-2′-deoxyuridine (EdU), cycle, and transwell assays. The effects of WSB2 knockdown on melanoma in vivo were determined using a xenograft mouse model. To investigate the underlying mechanisms, levels of c-Myc, β-catenin, phosphorylated retinoblastoma (p-Rb), cyclin-dependent kinase 4 (CDK4), and Cyclin D3 proteins were determined by western blotting in melanoma A375 cells with WSB2 knocked down. Furthermore, β-catenin agonism, SKL2001, was used to evaluate the mechanisms by which knockdown of WSB2 regulated cell proliferation. ResultsWSB2 levels were high and they were associated with clinicopathological features in patients with melanoma. shRNA-mediated knockdown of WSB2 could inhibit proliferation, both in vivo and in vitro. Cycle progression and migration of A375 and G361 cells were also significantly inhibited by WSB2 knockdown. Moreover, down-regulation of WSB2 decreased the levels of c-Myc, β-catenin, p-Rb, Cyclin-dependent kinase 4 (CDK4), and Cyclin D3 in melanoma G361 cells with WSB2 knocked down. Moreover, SKL2001 could effectively rescue WSB2 knockdown-mediated inhibition of cell proliferation in melanoma. ConclusionThis is the first report to demonstrate the effects of WSB2 on melanoma cell function. WSB2 has potential to become a new therapeutic target in patients with melanoma.

NEAT1 promotes retinoblastoma progression via modulating miR-124.

The long noncoding RNA nuclear‐enriched abundant transcript 1 (NEAT1) is reportedly involved in the initiation and progression of cancers of several types. However, the role, expression status, and the detailed mechanism of NEAT1 in retinoblastoma (RB) yet need to be unraveled. We explored the role and the mechanism of NEAT1 activity in RB. Our data show enhanced NEAT1 expression in RB‐affected tissues compared with the corresponding control. Functional experiments reveal that a NEAT1 knockdown in RB cells significantly inhibits proliferation, cycle progression, and facilitates apoptosis and caspase‐3 and ‐9 activities. Besides that, miR‐124 was predicted to be a target of NEAT1 and its reduced expression, as well as the inverse correlation of NEAT1 with miR‐124, was observed in RB‐affected tissues. Further, luciferase and RNA immunoprecipitation (RIP) assays confirmed the interaction between NEAT1 and miR‐124. Rescue experiments confirmed that the inhibition of miR‐124 could reverse the effect of NEAT1 on RB cell proliferation, cycle arrest, apoptosis, and caspase‐3 and ‐9 activities. Thus, NEAT1 promotes RB progression by sponging miR‐124, providing a therapeutic target for RB.

Phase Dynamics on Conversion-Reaction-Based Tin-Doped Ferrite Anode for Next-Generation Lithium Batteries.

The conventional view of conversion reaction is based on the reversibility, returning to an initial material structure through reverse reaction at each cycle in cycle life, which impedes the complete understanding on a working mechanism upon a progression of cycles in conversion-reaction-based battery electrodes. Herein, a series of tin-doped ferrites (Fe3- xSn xO4, x = 0-0.36) are prepared and applied to a lithium-ion battery anode. By achieving the ideal reoxidation into SnO2, the Fe2.76Sn0.24O4 composite anchored on reduced graphene oxide shows a high reversible capacity of 1428 mAh g-1 at 200 mA g-1 after 100 cycles, which is the best performance of Sn-based anode materials so far. Significantly, a newly formed γ-FeOOH phase after 100 cycles is identified from topological features through synchrotron X-ray absorption spectroscopy with electronic and atomic structural information, suggesting the phase transformation from magnetite to lepidocrocite upon cycling. Contrary to the conventional view, our work suggests a variable working mechanism in an iron-based composite with the dynamic phases from iron oxide to iron oxyhydroxide in the battery cycle life, based on the reactivity of metal nanoparticles formed during reaction toward the solid electrolyte interface layer.

Golgi Oncoprotein GOLPH3 Gene Expression Is Regulated by Functional E2F and CREB/ATF Promoter Elements.

The Golgi organelle duplicates its protein and lipid content to segregate evenly between two daughter cells after mitosis. However, how Golgi biogenesis is regulated during interphase remains largely unknown. Here we show that messenger RNA (mRNA) expression of GOLPH3 and GOLGA2, two genes encoding Golgi proteins, is induced specifically in G1 phase, suggesting a link between cell cycle regulation and Golgi growth. We have examined the role of E2F transcription factors, critical regulators of G1 to S progression of the cell cycle, in the expression of Golgi proteins during interphase. We show that promoter activity for GOLPH3, a Golgi protein that is also oncogenic, is induced by E2F1-3 and repressed by E2F7. Mutation of the E2F motifs present in the GOLPH3 promoter region abrogates E2F1-mediated induction of a GOLPH3 luciferase reporter construct. Furthermore, we identify a critical CREB/ATF element in the GOLPH3 promoter that is required for its steady state and ATF2-induced expression. Interestingly, depletion of GOLPH3 with small interfering RNA (siRNA) delays the G1 to S transition in synchronized U2OS cells. Taken together, our results reveal a link between cell cycle regulation and Golgi function, and suggest that E2F-mediated regulation of Golgi genes is required for the timely progression of the cell cycle.

Virtual Screening, Molecular Modelling and Biochemical Studies to Exploit PF14_0660 as a Target to Identify Novel Anti-malarials

Background: Malaria Parasite relies heavily on signal transduction pathways to control growth, the progression of the life cycle and sustaining stress for its survival. Unlike kinases, Plasmodium's phosphatome is one of the smallest and least explored for identifying drug target for clinical intervention. PF14_0660 is a putative protein present on the chromosome 14 of Plasmodium falciparum genome. Methods: Multiple sequence alignment of PF14_0660 with other known protein phosphatase indicate the presence of phosphatase motif with specific residues essential for metal binding, catalysis and providing structural stability. PF14_0660 is a mixed α/β type of protein with several β -sheet and α-helix arranged to form βαβαβα sub-structure. The surface properties of PF14_0660 is conserved with another phosphate of this family, but it profoundly diverges from the host protein tyrosine phosphatase. PF14_0660 was cloned, over-expressed and protein is exhibiting phosphatase activity in a dose-dependent manner. Docking of Heterocyclic compounds from chemical libraries into the PF14_0660 active site found nice fitting of several candidate molecules. Results: Compound PPinh6, PPinh 7 and PPinh 5 are exhibiting antimalarial activity with an IC50 of 1.4 ± 0.2µM, 3.8 ± 0.3 µM and 9.4 ± 0.6&#181M respectively. Compound PPinh 6 and PPinh 7 are inhibiting intracellular PF14_0660 phosphatase activity and killing parasite through the generation of reactive oxygen species. Conclusion: Hence, a combination of molecular modelling, virtual screening and biochemical study allowed us to explore the potentials of PF14_0660 as a drug target to design anti-malarials.

Upregulated gga-miR-16-5p Inhibits the Proliferation Cycle and Promotes the Apoptosis of MG-Infected DF-1 Cells by Repressing PIK3R1-Mediated the PI3K/Akt/NF-κB Pathway to Exert Anti-Inflammatory Effect.

Mycoplasma gallisepticum (MG) mainly infects chickens to initiate chronic respiratory disease (CRD). microRNAs (miRNAs) play vital roles according to previously reported studies. Our previous study showed that gga-miR-16-5p, in MG-infected lungs of chicken embryo, was upregulated by Illumina sequencing. The study aimed to reveal what role gga-miR-16-5p plays in CRD progression. gga-miR-16-5p was upregulated in MG-infected fibroblast cells (DF-1). Phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1) was demonstrated as the target gene of gga-miR-16-5p. Furthermore, PIK3R1 expression was lower in MG-infected groups than it in noninfected controls measured by qPCR. Additionally, overexpressed gga-miR-16-5p could downregulate PIK3R1 and phosphorylated serine/threonine kinase (p-Akt) to express protein, whereas there is an opposite effect on inhibition. Overexpressed gga-miR-16-5p resulted in decreased activity of tumor necrosis factor alpha (TNF-α) and the nuclear factor-kappaB (NF-κB) by qPCR. Furthermore, overexpressed gga-miR-16-5p restricted cell multiplication, cycle progression, and increased apoptosis of MG-infected DF-1 cells, whereas inhibited gga-miR-16-5p led to the opposite effect. Collectively, upregulated gga-miR-16-5p could decrease multiplication, cycle progression, and increase apoptosis of MG-infected DF-1 cells, at least partly through directly targeting PIK3R1 and inhibiting PI3K/Akt/NF-κB pathway to exert an anti-inflammatory effect. Our results will provide more experimental evidence to bring pathogenesis of MG infection to light.

Abstract P4-07-07: Triple targeted combination therapy with tucatinib, palbociclib and fulvestrant is active in hormone receptor and HER2-positive human breast tumor cell lines

Abstract Hormone receptor and HER2-positive (HR+/HER2+) breast cancer displays increased resistance to anti-hormonal and HER2-targeted agents. HER2 and estrogen receptor signaling converge at cyclin D1 and CDK4/6 complex, resulting in the increased cyclin D1 expression and accelerated progression of the cell cycle. Therefore, triple blockade of the estrogen receptor, HER2 and CDK4/6 is reasonable as a novel approach to treatment of HR+/HER2+ disease. Here for the first time we report high activity of novel combination therapy with HER2-targeted small molecule inhibitor tucatinib, CDK4/6 inhibitor palbociclib, and selective estrogen receptor blocker fulvestrant in human breast tumor cell lines. Methods: HR+/HER2+ human breast tumor cell lines BT474, MDA-MB-361 and UACC-812 were cultured in standard conditions. The subclones of BT474 and MDA-MB-361 resistant to tucatinib and palbociclib were generated by culturing cells in the increasing concentrations of inhibitors over 6 months. Cell survival assays were performed with Cell Titer Glow (Promega, Madison, WI) after 72 hrs of treatment with vehicle, single agents tucatinib, palbociclib and fulvestrant, and with dual and triple combinations. Tucatinib was provided by Seattle Genetics (Seattle, WA), palbociclib and fulvestrant were purchased from Selleckchem (Houston, TX). Protein lysates for western blot were prepared after 24, 48 or 72 hrs of drug treatment at IC70 using RIPA lysis buffer (Thermo Fisher, Waltham, MA) containing protease/phosphatase inhibitors (Roche Diagnostic, Indianapolis, IN). Signals were quantified with NIH ImageJ Imaging Analysis Software or Odyssey Imager software (Li-Cor Bioscience, Lincoln, NE) and normalized to vinculin or alpha-tubulin. Results: Dual combinations of tucatinib with fulvestrant and tucatinib with palbociclib were synergistic in all three HR+/HER2+ cell lines. Addition of a constant dose of fulvestrant to a combination of tucatinib and palbociclib further improved tumor cell killing in all three breast tumor cell lines tested. Combination of tucatinib and palbociclib resulted in greater suppression of phospho-AKT and total and phospho-CDK2 comparing to either agent alone. Treatment with single agents tucatinib and palbociclib lead to a significant increase of cyclin E expression at 48 and 72hrs after treatment, however, this compensatory mechanism of cell cycle progression was not observed after treatment with tucatinib and palbociclib combination. Experiments with the resistant BT474 and MDA-MB-361 subclones showed that cross-treatment of cells resistant to palbociclib with tucatinib and vice versa suppressed cell growth and decreased expression of phospho-ERK1/2, phospho-AKT, phospho-CDK2 and cyclin E. Conclusions: In HR+/HER2+ human breast tumor cell lines combination therapy with tucatinib, palbociclib and fulvestrant demonstrated significant synergy, potentially acting through suppression of phospho-AKT and cyclin E pathways, therefore suggesting a promising novel approach to treatment of HR+/HER2+ breast cancer. Clinical testing of tucatinib, palbociclib and anti-hormonal therapy in patients with HR+/HER2+ metastatic breast cancer is ongoing (phase Ib/II clinical trial NCT03054363). Citation Format: Shagisultanova E, Borakove M, Kabos P, Borges V. Triple targeted combination therapy with tucatinib, palbociclib and fulvestrant is active in hormone receptor and HER2-positive human breast tumor cell lines [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P4-07-07.

Semi-Intact Cell System for Reconstituting and Analyzing Cellular Golgi Dynamics.

Morphology of Golgi apparatus changes frequently and diversely depending on various cellular conditions and these changes correlate with the balance between membrane inflow and outflow at the Golgi via vesicular transports. In a previous study, we introduced a semi-intact cell system suitable for the reconstitution of morphological changes that organelles undergo as well as the vesicular transport between them. Semi-intact cells are cells that have undergone plasma membrane permeabilization by the cholesterol-dependent pore-forming cytolysin, streptolysin O (SLO). Permeabilization enables the introduction of various molecules into the cells, as well as the substitution of the original cytosol with an exogenously made cytosol prepared from cells in various stages of cell cycle, differentiation, and disease progression. Coupled with a green fluorescent protein(GFP)-visualization technique, this cell-based system enables the analysis of the molecular mechanisms underlying biological processes that are highly dependent on the integrity of the intracellular architecture. In this chapter, we present a variety of reconstitution assays concerning biological reactions pertaining to the Golgi apparatus.

The oncoprotein HBXIP – its functions and roles in oncogenesis

Nowadays, Hepatitis B X interacting protein (HBXIP) is an object of scientists’ interest worldwide. It is a protein with significant involvement in the development of malignant tumors like breast or ovarian cancer. One of the most important functions of HBXIP is the regulation of cell proliferation, which is related to the progression of a cell cycle. Many studies provide the growing number of evidence that HBXIP plays various important roles, including the regulation of a cell cycle through complexes with survivin, belonging to the inhibitors of apoptosis and interactions with transcriptional factors like STAT4, SP1, TFIID or E2F1. It also has the influence on the promotion of tumor angiogenesis thanks to the association with VEGF and FGF8. Another important role of HBXIP is a reprogramming of glucose metabolism to conditions favorable to growing cancerous cells due to regulating the activation of SCO2 and PDHA1. Furthermore, it impacts on the complement-dependent cytotoxicity, also, HBXIP affects on lipid metabolism through disturbing of metabolic pathways of FAS. According to recent studies, HBXIP can be used as a prognostic biomarker in many tumors, including cervical cancer, ovarian cancer, and esophageal squamous cell carcinoma thanks to the high expression of this protein noted exclusively in these tumor tissues. What is even more interesting, it significantly correlates with clinical attributes like metastasis to lymph nodes or grading and in some cases can potentially be used as the indicator of prognosis of treatment effectiveness. The paper is review through main functions of HBXIP and its possible applications.

A novel and safe small molecule enhances hair follicle regeneration by facilitating metabolic reprogramming

Targeting hair follicle regeneration has been investigated for the treatment of hair loss, and fundamental studies investigating stem cells and their niche have been described. However, knowledge of stem cell metabolism and the specific regulation of bioenergetics during the hair regeneration process is currently insufficient. Here, we report the hair regrowth-promoting effect of a newly synthesized novel small molecule, IM176OUT05 (IM), which activates stem cell metabolism. IM facilitated stemness induction and maintenance during an induced pluripotent stem cell generation process. IM treatment mildly inhibited mitochondrial oxidative phosphorylation and concurrently increased glycolysis, which accelerated stemness induction during the early phase of reprogramming. More importantly, the topical application of IM accelerated hair follicle regeneration by stimulating the progression of the hair follicle cycle to the anagen phase and increased the hair follicle number in mice. Furthermore, the stem cell population with a glycolytic metabotype appeared slightly earlier in the IM-treated mice. Stem cell and niche signaling involved in the hair regeneration process was also activated by the IM treatment during the early phase of hair follicle regeneration. Overall, these results show that the novel small molecule IM promotes tissue regeneration, specifically in hair regrowth, by restructuring the metabolic configuration of stem cells.

LncRNA LINC00968 accelerates the proliferation and fibrosis of diabetic nephropathy by epigenetically repressing p21 via recruiting EZH2

Emerging evidence have indicated the vital roles of long noncoding RNAs (lncRNAs) in the pathophysiological process of diabetic nephropathy. However, the deepgoing mechanism that lncRNAs regulate the diabetic nephropathy pathogenesis is still ambiguous. In present study, we found that lncRNA LINC00968 expression was high-expressed in the diabetic db/db mouse tissue and high-glucose induced mesangial cells. Functional experiments indicated that LINC00968 silencing by siRNAs significantly inhibited the proliferation and cycle progression, and decreased the extracellular matrix (ECM) proteins (fibronectin, collagen IV) expression in the high glucose induced of mesangial cells. RNA immunoprecipitation (RIP) and chromatin immunoprecipitation (ChIP) assay revealed that LINC00968 recruit EZH2 to the promoter of p21 to inhibit its expression. In summary, our results support the conclusion that lncRNA LINC00968 accelerates the proliferation and fibrosis of mesangial cells by epigenetically repressing p21 via recruiting EZH2, providing a novel insight for the diabetic nephropathy pathogenesis.

Effect of Crystal Orientation of Cu Current Collectors on Cycling Stability of Li Metal Anodes

Li metal anode is an attractive material for secondary batteries due to its high capacity of 3860 mAh/g. However, Li metal is well known for its hazardous behavior with air and moisture. Therefore, reducing the amount of Li metal in anodes is effective not only increasing energy density but also guarantee the safety of the batteries. For these reason, anode-free lithium batteries(AFLBs), in which Li electrodeposits onto Cu current collector, has been proposed by in-situ formation of Li metal1. Despite the potential increase in energy density of AFLBs comparing to conventional Li-ion batteries2, low coulombic efficiency due to non-uniform deposition nature of Li metal prevents AFLBs for practical applications. To address this issue, many methods such as high concentrated electrolyte, formation of protective layer, and electrolyte additives are developed, but AFLBs are still not realized. On the other hand, our previous work3 revealed that shape of Li precipitates is influenced by crystal orientation of Cu current collectors. This fact suggests that cycling stability of Li metal anode is also affected by the orientation of the Cu collector. Thus, in the present work, we performed cyclic voltammetry to investigate the transition of the characteristics. The experiments of Li electrodeposition and stripping on single crystal Cu current collectors were conducted in a three-pole cell. The orientation of Cu are (111), (101), and (001). Counter and reference electrodes are both Li metal. Prior to experiments, the crystal orientation of collectors was analyzed by electron backscatter diffraction (EBSD). An electrolyte is consisted of 1 M LiPF6 in EC:EMC=5:5. Li was deposited at a current density of 0.5 mA/cm2 with charge capacity of 0.1 mA/cm2. Subsequently, the Li was stripped at same current density with 0.05 mA/cm2. Cyclic voltammetry was performed at a scan rate of 5.0 mV/s. SEM analysis was also conducted to examine Li morphology. Figure 1 shows cyclic voltammograms of Li deposition and stripping on various Cu collectors. Negative and positive peaks represent Li deposition and stripping respectively. Each peak current decreases substantially as cycle proceeds, probably because of the development of the resistive SEI layer. The smallest overpotential for the first Li deposition was observed in Cu(111), followed by Cu(101) and Cu(001). Uniform formation of the resistive layer expectedly causes this since the SEM analysis revealed that Li morphology on Cu(111) is the most homogeneous. Moreover, Cu(111) delivered the lowest and the most stable overpotential over 10 cycle, while those in the other cases are larger and increase as the cycle progress. Therefore, it can we concluded that crystal orientation of collectors is an important factor for Li metal batteries and Cu(111) is effective for high cycle stability.

Abstract 3488: Elucidation of Warburg effect and cell cycle regulations through analysis of cancer tissue omics data and kinetic modeling

Abstract Cancer cells tend to depend more on anerobic glycolysis to generate adenosine 5'- triphosphate (ATP), which is an inefficient energy producing process termed as “The Warburg effect”. The causes or functions of Warburg effect have been extensively studied, however, the growth advantage it confers to cancer cells still remains unclear. In this study, we propose that the metabolic shift in cancer cells would facilitate the accumulation of nutrients and biomass (e.g., nucleotides, amino acids and lipids), which would promote the cell cycle initiation and progression as unicellular organisms do. Specifically, by comprehensively analyze the gene expression and mutational profile in 16 cancer types, we identified (1) the gene expression profile of cancer cells is evolutionarily more unicellular like when compared to normal; and (2) certain cell cycle associated mutations are associated with the metabolic alternations in cancer cells, such as nucleotide synthesis pathways and lipids metabolism. Moreover, we developed a system of kinetic equations to capture the metabolic relations between nutrient metabolism and cell cycle initiation and progression. The kinetic model shows that the rapid nucleotide synthesis as well as high influx of glutamate would affect the activities of cell cycle signaling and CDKs genes. Our study provides a novel mechanistic view in linking the Warburg effect and cancer cell cycle initiation and progression. Citation Format: Tao Sheng. Elucidation of Warburg effect and cell cycle regulations through analysis of cancer tissue omics data and kinetic modeling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3488.

Inhibition of Hornification of Eucalyptus Kraft Pulp by Acetylation

Bleached eucalyptus Kraft pulp was acetylated with acetic anhydride under mild conditions, and recycling treatments were performed for acetylated pulp and unmodified pulp. The results indicated that Kraft pulp could be acetylated with acetic anhydride in absence of the catalyst, and hydrophobic acetyl functional groups were introduced into cellulosic fibres. Acetylation can restored the swelling ability of Kraft fibres, inhibit the hornification of Kraft pulp and improve pulp performance with the progress of cycles.

Abstract 2814: Antitumor effects of WEE1 inhibitor, AZD1775 alone or in combination with PARP inhibitor in triple negative breast cancer cells

Abstract Background: Wee1 is a critical component of the G2/M cell cycle checkpoint control and mediates cell cycle arrest by regulating the phosphorylation of CDC2. Inhibition of Wee1 has been reported to enhance the cytotoxic effect of DNA damaging agents in different types of carcinomas. Especially, recent reports have shown that DNA replication stress and double strand breaks are induced by the activation of CDK1 following WEE1 inhibitor treatment. We investigated the effects and underlying mechanisms of WEE1 inhibitor, AZD1775, in Triple Negative Breast Cancer (TNBC). In addition, we examined whether AZD1775 would enhance the anti-tumor effect of PARP inhibitor in TNBC cells by modulating DNA repair activity. Methods: MTT assay was performed to investigate the antitumor effect of WEE1 inhibitor, AZD1775 alone or with PARP inhibitor in breast cancer cell lines. The regulation of DNA damage response activity by AZD1775 or/and PARP inhibitor was accessed by the comet and western blotting. The status of the S phase was evaluated by EdU-BrdU dual pulse labeling, and the drug effect was also confirmed in vivo by mouse xenograft model of TNBC. Results: AZD1775 showed heterogeneous antitumor effects among TNBC cells regardless of TP53 mutation statues. WEE1 inhibition led to accelerated S phase progression and mitotic entry in the sensitive MDA-MB-231 cells. In addition, increased DNA damage and caspase-3 dependent apoptosis were observed in sensitive cells following AZD1775 treatment. Furthermore, AZD1775 enhanced cellular sensitivity to PARP inhibitor through increase apoptosis and DNA damage accumulation. In a MDA-MB-231 xenograft model, AZD1775 impedes tumor growth as well. Conclusions: In the TNBC, the WEE1 inhibitor has a heterogeneous cytotoxic effect regardless of the p53 mutation status. In MDA-MB-231 cell, which is a sensitive TNBC cells to WEE1 inhibitor AZD1775, the progression of the cell cycle was accelerated and the aberrant DNA content was increased. This resulted in DNA damage, which led to cell death, and this effect could be applied in vivo. In addition, we investigated that WEE1 inhibition can increase PARP inhibitor sensitivity in TNBC cells via increasing accumulation of DNA damage. Our results provide a rationale for the future clinical trials of PARP inhibitor combined with WEE1 inhibition in triple negative breast cancer. Citation Format: Dong Hyeon Ha, Arrum Min, Seongyeong Kim, So Hyeon Kim, Hyemin Jang, Yu Jin Kim, Daeun Jung, YoonJung Park, Koung Jin Suh, Kyung-Hun Lee, Tae-Yong Kiim Kim, Seock-Ah Im. Antitumor effects of WEE1 inhibitor, AZD1775 alone or in combination with PARP inhibitor in triple negative breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2814.