Regulation of tumor cell migration by protein tyrosine phosphatase (PTP)-proline-, glutamate-, serine-,and threonine-rich sequence(PEST)

Protein tyrosine phosphatase (PTP)-proline-,glutamate-,serine-,and threonine-rich sequence (PEST) is ubiquitously expressed and is a critical regulator of cell adhesion and migration.PTP-PEST activity can be regulated transcriptionally via gene deletion or mutation in several types of human cancers or via post-translational modifications,including phosphorylation,oxidation,and caspase-dependent cleavage.PTP-PEST interacts with and dephosphorylates cytoskeletal and focal adhesion-associated proteins.Dephos-phorylation of PTP-PEST substrates regulates their enzymatic activities and/or their interaction with other proteins and plays an essential role in the tumor cell migration process.

Comparison of genetic impact on growth and wood traits between seedlings and clones from the same plus trees of Pinus koraiensis

To evaluate the relationships among clones and open pollinated families from the same plus trees and to select elite breeding materials,growth,and wood characteristics of 33-year-old Pinus korainsis clones and families were measured and analyzed.The results show that growth and wood characters varied significantly.The variation due to clonal effects was higher than that of family effects.The ratio of genetic to phenotypic coefficient of variation of clones in growth and wood traits was above 90%,and the repeatability of these characteristics was more than 0.8,whereas the ratio of genetic to phenotypic coefficient of variation of families was above 90%.The broad-sense heritability of all characteristics exceeded 0.4,and the narrow-sense family heritability of growth traits was less than 0.3.Growth characteristics were positively correlated with each other,but most wood properties were weakly correlated in both clones and families.Fiber length and width were positively correlated between clones and families.Using the membership function method,eleven clones and four families were selected as superior material for improved diameter growth and wood production,and two families from clonal and open-pollinated trees showed consistently better performance.Generally,selection of the best clones is an effective alternative to deployment of families as the repeatability estimates from clonal trees were higher than narrow-sense heritability estimates from open pollinated families.The results provide valuable insight for improving P.koraiensis breeding programs and subsequent genetic improvement.

Study on the physiological indices of Pinus sibirica and Pinus koraiensis seedlings under cold stress

Although Pinus sibirica and Pinus koraiensis are resistant to cold or low temperatures in the cold temperate regions of the northern hemisphere,the former has a stronger cold resistance.Research has been limited to the comparison of physiological responses of the two species to cold stress.In this study,5-year-old seedlings of P.sibirica and P.koraiensis were subjected to six temperature treatments,[20℃(control),0℃,-20℃,-40℃,-60℃,and-80℃],under different stress periods(6,12,24,and 48 h).The results showed that differences in each physiological index were significant between P.sibirica and P.koraiensis,except for the permeability of cell membranes,reactive oxygen species,proline and soluble proteins.An ANOVA test indicated that there were extreme differences among the temperatures for each index,stress time and temperature 9 time for most indices.All indices showed a similar trend for P.sibirica and P.koraiensis with decreasing temperature or the extension of stress time.Soluble sugars and proline increased at 0 to-20℃and then remained unchanged with temperature decline.Other indices showed an increase from 20 to-20℃,stable from-20 to-40℃and a decrease from-40 to-80℃.All the indices increased and then declined along with the prolonged cold stress time,except for the control.From 0 to-40℃,the permeability of cell membranes,relative conductivity,reactive oxygen species and malonaldehyde of P.koraiensis seedlings were higher than in P.sibirica,but superoxide dismutase,peroxidase,catalase activity and soluble sugars,proline,and soluble proteins content emerged as opposite.This study compared the physiological mechanism responses to cold stress between P.sibirica and P.koraiensis to provide the basis for the introduction,distribution,and genetic improvement of these coniferous species.

Nonmetabolic functions of pyruvate kinase isoform M2 in controlling cell cycle progression and tumorigenesis

Pyruvate kinase catalyzes the rate-limiting final step of glycolysis, generating adenosine triphosphate (ATP) and pyruvate. The M2 tumor-specific isoform of pyruvate kinase (PKM2) promotes glucose uptake and lactate production in the presence of oxygen, known as aerobic glycolysis or the Warburg effect. As recently reported in Nature, PKM2, besides its metabolic function, has a nonmetabolic function in the direct control of cell cycle progression by activating β-catenin and inducing expression of the β-catenin downstream gene CCND1 (encoding for cyclin D1). This nonmetabolic function of PKM2 is essential for epidermal growth factor receptor (EGFR) activation-induced tumorigenesis.

Hybrid classification of coal and biomass by laser-induced breakdown spectroscopy combined with K-means and SVM

Laser-induced breakdown spectroscopy(LIBS) is a new technology suitable for classification of various materials. This paper proposes a hybrid classification scheme for coal, municipal sludge and biomass by using LIBS combined with K-means and support vector machine(SVM)algorithm. In the study, 10 samples were classified in 3 groups without supervision by K-means clustering, then a further supervised classification of 6 kinds of biomass samples by SVM was carried out. The results show that the comprehensive accuracy of the hybrid classification model is over 98%. In comparison with the single SVM classification model, the hybrid classification model can save 58.92% of operation time while guaranteeing the accuracy. The results demonstrate that the hybrid classification model is able to make an efficient, fast and accurate classification of coal, municipal sludge and biomass, furthermore, it is precise for the detection of various kinds of biomass fuel.

Analyzing of mixing performance determination factors for the structure of radial multiple jets-in-crossflow

The radial multiple jets-in-crossflow mixing structure(RMJCMS) is extensively used in industrial manufacture. In this research, the effects of thickness of injection ring on mixing performance and factors influencing the mixing performance of RMJCMS were discussed based on the results of computational fluid dynamics. The simulation results showed that the dimensionless mixing distance, with the increase of the thickness of injection ring, drops from 1.1 to 0.18 first and then increases to 0.27 while the uniformity of flux monotonously improves, manifesting that the consistency of flux is not the single element determining the mixing performance. Analyzing the simulation results, a conclusion was drawn that the consistency of flux, penetration mode and interaction among injection flows which can be altered by adjusting the thickness of injection ring, determine the mixing performance of RMJCMS jointly. That is to say, in RMJCMS an injection ring with a suitable thickness can realize the function of injection and rectification simultaneously, which not only improves the mixing performance but also reduces the complexity of RMJCMS as well.

PGK1-coupled HSP90 stabilizes GSK3βexpression to regulate the stemness of breast cancer stem cells

Objective:Glycogen synthase kinase-3β(GSK3β)has been recognized as a suppressor of Wnt/β-catenin signaling,which is critical for the stemness maintenance of breast cancer stem cells.However,the regulatory mechanisms of GSK3βprotein expression remain elusive.Methods:Co-immunoprecipitation and mass spectral assays were performed to identify molecules binding to GSK3β,and to characterize the interactions of GSK3β,heat shock protein 90(Hsp90),and co-chaperones.The role of PGK1 in Hsp90 chaperoning GSK3βwas evaluated by constructing 293T cells stably expressing different domains/mutants of Hsp90α,and by performing a series of binding assays with bacterially purified proteins and clinical specimens.The influences of Hsp90 inhibitors on breast cancer stem cell stemness were investigated by Western blot and mammosphere formation assays.Results:We showed that GSK3βwas a client protein of Hsp90.Hsp90,which did not directly bind to GSK3β,interacted with phosphoglycerate kinase 1 via its C-terminal domain,thereby facilitating the binding of GSK3βto Hsp90.GSK3β-bound PGK1 interacted with Hsp90 in the“closed”conformation and stabilized GSK3βexpression in an Hsp90 activity-dependent manner.The Hsp90 inhibitor,17-AAG,rather than HDN-1,disrupted the interaction between Hsp90 and PGK1,and reduced GSK3βexpression,resulting in significantly reduced inhibition ofβ-catenin expression,to maintain the stemness of breast cancer stem cells.Conclusions:Our findings identified a novel regulatory mechanism of GSK3βexpression involving metabolic enzyme PGK1-coupled Hsp90,and highlighted the potential for more effective cancer treatment by selecting Hsp90 inhibitors that do not affect PGK1-regulated GSK3βexpression.

Activation of the complement system sensitizes immune checkpoint blockade

1.Introduction.ecently,a study published in Nature Cancer by Shao et al.1 shows that intratumoral activation of the complement system promotes antitu-mor immunity and eliminates epidermal growth factor receptor(EGFR)-activated lung cancer resistance to immune-checkpoint inhibitor(ICI)treatment.This study unveiled the molecular and genetic mechanism underlying the EGFR-elicited intratumoral complement inhibition and provided a novel strategy to treat lung cancer with combination of ICIs and intratumoral complement system activation.

A non-metabolic function of hexokinase 2 in small cell lung cancer:promotes cancer cell stemness by increasing USP11-mediated CD133 stability

Background:Maintenance of cancer stem-like cell(CSC)stemness supported by aberrantly regulated cancer cell metabolism is critical for CSC self-renewal and tumor progression.As a key glycolytic enzyme,hexokinase 2(HK2)plays an instrumental role in aerobic glycolysis and tumor progression.However,whether HK2 directly contribute to CSC stemness maintenance in small cell lung cancer(SCLC)is largely unclear.In this study,we aimed to investgate whether HK2 independent of its glycolytic activity is directly involved in stemness maintenance of CSC in SCLC.Methods:Immunoblotting analyses were conducted to determine the expression of HK2 in SCLC CSCs and their differentiated counterparts.CSC-like properties and tumorigenesis of SCLC cells with or without HK2 depletion or overexpression were examined by sphere formation assay and xenograft mouse model.Immunoprecipitation and mass spectrometry analyses were performed to identify the binding proteins of CD133.The expression levels of CD133-associated and CSC-relevant proteins were evaluated by immunoblotting,immunoprecipitation,immunofluorescence,and immunohistochemistry assay.RNA expression levels of Nanog,POU5F1,Lin28,HK2,Prominin-1 were analyzed through quantitative reverse transcription PCR.Polyubiquitination of CD133 was examined by in vitro or in vivo ubiquitination assay.CD133+cells were sorted by flow cytometry using an anti-CD133 antibody.Results:We demonstrated that HK2 expression was much higher in CSCs of SCLC than in their differentiated counterparts.HK2 depletion inhibited CSC stemness and promoted CSC differentiation.Mechanistically,nonmitochondrial HK2 directly interacted with CD133 and enhanced CD133 expression without affecting CD133 mRNA levels.The interaction of HK2 and CD133 promoted the binding of the deubiquitinase ubiquitin-specific protease 11(USP11)to CD133,thereby inhibiting CD133 polyubiquitylation and degradation.HK2-mediated upregulation of CD133 expression enhanced the expression of cell renewal regulators,SCLC cell stemness,and tumor growth in mice.In addition,HK2 expression was positively correlated with CD133 expression in human SCLC specimens,and their expression levels were associated with poor prognosis of SCLC patients.Conclusions:These results revealed a critical non-metabolic function of HK2 in promotion of cancer cell stemness.Our findings provided new insights into the multifaceted roles of HK2 in tumor development.

Governing glutaminolysis by regulation of glutaminase succinylation

Of the 20 ribosomally coded amino acid residues,lysine(K)can be frequently and uniquely modified by many types of posttranslational acylation,including acetylation,succinylation,crotonylation,propionylation,butyrylation,glutarylation,malonylation,β-hydroxybutyrylation,and 2-hydroxyisobutyrylation in histones and other proteins(Li et al.,2018a;Xu et al.,2021).Among these acylations,succinylation is a commonly occurred modification,and histone H3K79 succinylation is identified in the transcription start sites of more than 7,000 genes,implying its critical role in the expression regulation of a large number of genes(Wang et al.,2017).Protein succinylation requires the highenergy metabolite,succinyl-coenzyme A(CoA).

Genome-wide CRISPR screening identifies critical role of phosphatase and tensin homologous (PTEN) in sensitivity of acute myeloid leukemia to chemotherapy

Although significant progress has been made in the development of novel targeted drugs for the treatment of acute myeloid leukemia(AML)in recent years,chemotherapy still remains the mainstay of treatment and the overall survival is poor in most patients.Here,we demonstrated the antileukemia activity of a novel small molecular compound NL101,which is formed through the modification on bendamustine with a suberanilohydroxamic acid(SAHA)radical.NL101 suppresses the proliferation of myeloid malignancy cells and primary AML cells.It induces DNA damage and caspase 3-mediated apoptosis.A genome-wide clustered regularly interspaced short palindromic repeats(CRISPR)library screen revealed that phosphatase and tensin homologous(PTEN)gene is critical for the regulation of cell survival upon NL101 treatment.The knockout or inhibition of PTEN significantly reduced NL101-induced apoptosis in AML and myelodysplastic syndrome(MDS)cells,accompanied by the activation of protein kinase B(AKT)signaling pathway.The inhibition of mammalian target of rapamycin(mTOR)by rapamycin enhanced the sensitivity of AML cells to NL101-induced cell death.These findings uncover PTEN protein expression as a major determinant of chemosensitivity to NL101 and provide a novel strategy to treat AML with the combination of NL101 and rapamycin.

Molecular Basis of KAT2A Selecting Acyl-CoA Cofactors for Histone Modifications

Emerging discoveries about undocumented acyltransferase activities of known histone acetyltransferases(HATs)advance our understandings in the regulation of histone modifications.However,the molecular basis of HATs selecting acyl coenzyme A(acyl-CoA)substrates for histone modification is less known.We here report that lysine acetyltransferase 2A(KAT2A)as an illustrative instance of HATs can selectively utilize acetyl-CoA,propionyl-CoA,butyryl-CoA,and succinyl-CoA to directly deposit 18 histone acylation hallmarks in nucleosome.By analyzing the co-crystal structures of the catalytic domain of KAT2A in complex with acetyl-CoA,propionyl-CoA,butyryl-CoA,malonyl-CoA,succinyl-CoA,and glutaryl-CoA,we conclude that the alternative substrate-binding pocket of KAT2A and the length and electrostatic features of the acyl chain cooperatively determine the selection of the acyl-CoA substrates by KAT2A.This study reveals the molecular basis underlying the pluripotency of HATs that selectively install acylation hallmarks in nucleosomes,which might serve as instrumental mechanism to precisely regulate histone acylation profiles in cells.

Abrogation of nuclear entry of TERT by fructose 1,6-bisphosphatase 1-mediated dephosphorylation

Telomeres maintain chromosome integrity.Loss of telomere function,which is attributed to progressively shortened telomeres in each round of DNA replication,induces endto-end fusion of chromosomes,anaphase bridges with subsequent chromosome breakage,and eventually leads to senescence and apoptosis in normal cells[1].In cancer cells,highly activated telomerase synthesizes telomere repeats to promote telomere elongation.

Cancer metabolism and tumor microenvironment:fostering each other?

The changes associated with malignancy are not only in cancer cells but also in environment in which cancer cells live.Metabolic reprogramming supports tumor cells’high demand of biogenesis for their rapid proliferation,and helps tumor cells to survive under certain genetic or environmental stresses.Emerging evidence suggests that metabolic alteration is ultimately and tightly associated with genetic changes,in particular the dysregulation of key oncogenic and tumor suppressive signaling pathways.Cancer cells activate HIF signaling even in the presence of oxygen and in the absence of growth factor stimulation.This cancer metabolic phenotype,described firstly by German physiologist Otto Warburg,ensures enhanced glycolytic metabolism for the biosynthesis of macromolecules.The conception of metabolite signaling,i.e.,metabolites are regulators of cell signaling,provides novel insights into how reactive oxygen species(ROS)and other metabolites deregulation may regulate redox homeostasis,epigenetics,and proliferation of cancer cells.Moreover,the unveiling of noncanonical functions of metabolic enzymes,such as the moonlighting functions of phosphoglycerate kinase 1(PGK1),reassures the importance of metabolism in cancer development.The metabolic,microRNAs,and ncRNAs alterations in cancer cells can be sorted and delivered either to intercellular matrix or to cancer adjacent cells to shape cancer microenvironment via media such as exosome.Among them,cancer microenvironmental cells are immune cells which exert profound effects on cancer cells.Understanding of all these processes is a prerequisite for the development of a more effective strategy to contain cancers.

Metabolic features of cancer cells

Cancer cells uniquely reprogram their cellular activities to support their rapid proliferation and migration and to coun-teract metabolic and genotoxic stress during cancer progression.In this reprograming,cancer cells’metabolism and other cellular activities are integrated and mutually regulated,and cancer cells modulate metabolic enzymes spatially and temporally so that these enzymes not only have altered metabolic activities but also have modulated subcellular localization and gain non-canonical functions.This review and several others in this issue of Cancer Communications discuss these enzymes’newly acquired functions and the non-canonical functions of some metabolites as features of cancer cell metabolism,which play critical roles in various cellular activities,including gene expression,anabolism,catabolism,redox homeostasis,and DNA repair.

Associations of PGK1 promoter hypomethylation and PGK1-mediated PDHK1 phosphorylation with cancer stage and prognosis:a TCGA pan-cancer analysis

Background:Cancer cells reprogram metabolism for proliferation.Phosphoglycerate kinase 1(PGK1),as a glycolytic enzyme and newly identified protein kinase,coordinates glycolysis and mitochondrial metabolism.However,the clini-cal significance of PGK1 expression and function in cancer progression is unclear.Here,we investigated the relation-ship between the progression and prognosis of multiple cancer types and PGK1 expression and its function in the mitochondrial metabolism regulation.Methods:We performed pan-cancer analyses of PGK1 mRNA level and DNA methylation in 11,908 tumor tissues and 1582 paired normal tissues across 34 cancer types in The Cancer Genome Atlas datasets.Using specific antibodies against PGK1 S203 and PDHK1 T338 phosphorylation,we performed immunohistochemistry with tissue microarray assay in additional 818 cancer cases with 619 paired normal tissues from five cancer types.Results:The PGK1 mRNA level was significantly elevated with hypomethylation in promotor regions and associated with advanced TNM stage in 15 and four cancer types,respectively.In breast carcinoma,elevated PGK1 mRNA level and promoter hypomethylation were associated with poor prognosis.Positively correlated PGK1 S203 and PDHK1 T338 phosphorylation levels were significantly associated with short overall survival(OS)in cancers of the breast,liver,lung,stomach,and esophagus and with advanced TNM stage in breast and esophageal cancers.PGK1 pS203 and PDHK1 pT338 were also independent predictors of short OS in liver,lung,and stomach cancer.Conclusions:The elevated expression,promoter hypomethylation,and phosphorylation of PGK1 and PDHK1 were related with disease progression and short OS in diverse types of cancer.PGK1 and PDHK1 phosphorylation may be potential prognostic biomarkers.

Fructose and fructose kinase in cancer and other pathologies

Fructose metabolism and fructose kinase KHKe C/A are key factors in the development of lipid oversynthesis-promoted metabolic disorders and cancer.Here,we summarize and discuss the current knowledge about the specific features of fructose metabolism and the distinct roles of KHKe C and KHKe A in metabolic liver diseases and their relevant metabolic disorders and cancer,and we highlight the specific protein kinase activity of KHKe A in tumor development.In addition,different approaches that have been used to inhibit KHK and the exploration of KHK inhibitors in clinical treatment are introduced.

A newly discovered role ofmetabolic enzyme PCK1 as a protein kinase to promote cancer lipogenesis

Lipid metabolism,in particular fatty acid and cholesterol synthesis,is essential to convert nutrients into metabolic intermediates for membrane biosynthesis,energy storage and the generation of signaling molecules.Tumor cells maintain high level of lipid metabolism for rapid cell proliferation[1-4].Transcription of genes required for fatty acid and cholesterol synthesis and cholesterol uptake is controlled by membrane-bound transcription factor sterol regulatory element-binding proteins(SREBPs),including SREBP-1a,SREBP-1c/ADD1 and SREBP-2 isoforms[5].The function of SREBPs is mainly regulated by an escort protein(the SREBP cleavage-activating protein[SCAP])and endoplasmic reticulum(ER)anchor proteins(insulininduced genes[Insigs]),during the feedback loop of cholesterol synthesis[6-8].Under sterol-depleted conditions,SCAP and SREBPs complex is captured by COPIImediated vesicles and transported from the ER to the Golgi apparatus[7,9],where the SREBPs are proteolytically processed by Site-1 protease(S1P)and Site-2 protease(S2P)to yield active amino-terminal fragments that enter the nucleus for gene transcription[10,11].Under high intracellular sterol conditions,abundant cholesterol in the ER membrane binds to SCAP,induces its conformational change,and enables it to bind to Insigs.

Aerobic glycolysis promotes tumor immune evasion and tumor cell stemness through the noncanonical function of hexokinase 2

The Warburg effect is a key metabolic feature of cancer cells and manifests as substantially increased glycolysis regardless of the presence of oxygen[1].The initial step of glycolysis is catalyzed by hexokinase(HK),which converts glucose to the metabolic intermediate glucose-6-phosphate(G-6-P).Four kinds of HK isozymes,HK1,HK2,HK3 and HK4,have been found in mammals.HK1 and HK2 bind to the mitochondrial outer membrane through interac-tions with voltage-dependent anion channels(VDACs),and both of thesemolecules have a high affinity for glucose[2].HK2 overexpression is frequently detected in cancer cells,leading to enhanced aerobic glycolysis and tumorigenesis that renders HK2 an attractive target for cancer treatment[3].Important advances in research on cancer metabolism in recent decades have revealed thatmetabolic enzymes have non-metabolic functions,which play pivotal roles in tumor development and progression[4–6].Recent studies uncovered that HK2 directly promoted tumor immune evasion through the activity of a previously unidentified protein kinase and tumor cell stemness via upregulation of CD133 expression.

A moonlighting function of choline kinase alpha 2 in the initiation of lipid droplet lipolysis in cancer cells

Cancer cells in the tumormicroenvironment,where nutrient availability is consistently changing during tumor progression,harness lipid metabolism to support their proliferation,survival,migration,invasion,and metastasis[1].When nutrients in the tumor microenvironment are sufficient,lipids such as triglycerides,steryl esters,and retinyl esters are stored in lipid droplets,which are dynamic lipid-rich cellular organelles surrounded by a single layer of polar and amphipathic phospholipids and structural proteins in the perilipin(PLIN)family.