Effects of Heterologously Overexpressing PIP5K-Family Genes in Arabidopsis on Inflorescence Development

Castor is one of the top 10 oil crops in the world and has extremely valuable uses.Castor inflorescences directly affect yield,so the study of inflorescence development is very important in increasing castor yield.Our previous studies have shown that the PIP5K gene family(PIP5Ks)is associated with inflorescence development.In this study,to determine the function of each PIP5K gene in castor,a female Lm-type castor line,aLmAB2,was used to determine the relative expression levels of the PIP5Ks in castor inflorescences.Six PIP5K genes were heterologously overexpressed in Arabidopsis thaliana,the relative expression of each gene and the effect on plants was determined in A.thaliana,and the relationships among the PIP5Ks in castor were inferred.The expression levels of the PIP5Ks in the female Lm-type castor line aLmAB2 were analyzed.The relative expression levels of the PIP5K9 and PIP5K11 genes were high(p<0.05)in isofemale inflorescences,and those of PIP5K1,PIP5K2,PIP5K6,and PIP5K8 were high(p<0.05)in female inflorescences but low(p<0.05)in bisexual inflorescences.The PIP5Ks were heterologously overexpressed in A.thaliana,and T3-generation plants with stable genetic resistance,i.e.,AT-PIP5K^(+)plants(AT-PIP5K1^(+),AT-PIP5K2^(+),AT-PIP5K6^(+),AT-PIP5K8^(+),AT-PIP5K9^(+),and ATPIP5K11^(+) plants),were obtained.Biological tests of the AT-PIP5K+plants showed that the growth of the main stem was significantly delayed in AT-PIP5K+plants compared with Columbia wild-type(WT)A.thaliana plants;the PIP5K1 and PIP5K2 genes promoted lateral stem growth and flower and silique development;and the PIP5K6,PIP5K8,PIP5K9 and PIP5K11 genes inhibited lateral stem growth and flower and silique development.The correlations among PIP5Ks in castor suggest that there may be a synergistic relationship among PIP5K1,PIP5K2,and PIP5K6 in castor inflorescences,and PIP5K8,PIP5K9,and PIP5K11 are complementary to the other three genes.

The establishment of a stable PC-3 cell line overexpressing micro RNA-145

Objective To establish stable prostate cancer bone metastasis cell line overexpressing microRAN-145 (miR-145)for the study of the mechanism of miR-145 in bone metastasis.Methods pMSCV-miR-145 plasmids and retroviruses of pMSCV-vector

Cloning and Overexpressing Apo-phycoerythrocyanin α-subunit Gene of Mastigocladus laminosus in E.coli

By PCR method, apo phycoerythrocyanin α subunit gene (pecA) of Mastigocladus laminosus (M. laminosus) was amplified from its genomic DNA, and then cloned in pBluescript. The pecA gene was subcloned into the expression vector pGEMD, and then transformed into E.coli BL21 (DE3). After induction, a new protein of molecular weight 19×10 3 existing in inclusion body was overexpressed. The expressed product was confirmed to be apo phycoerythrocyanin α subunit by Dot ELISA.

Enhanced Production of Natural Carotenoids from Genetically Engineered Rhodobacter sphaeroides Overexpressing CrtA

Carotenoids act as precursors of vitamin A,antioxidants,enhancers of immunity,and are thus widely used in food and pharmaceutical industry.Microbial fermentation is one of the most important solutions for production of natural carotenoids.Rhodobacter sphaeroides is one of most promising bacteria employed for large scale production of carotenoids.In the present study,crtA located in the carotenoids biosynthesis pathway in R.sphaeroides was amplified by PCR.The overexpression vector pRKcrtA was constructed and subsequently transferred into R.sphaeroides,producing the genetically engineered strain R.sphaeroides 2.4.1/pRKcrtA overexpressing crtA.The carotenoid production from the genetically engineered strain was significantly increased.Fermentation procedure was optimized for further enhanced carotenoids production.

A novel TNKS/USP25 inhibitor blocks the Wnt pathway to overcome multi-drug resistance in TNKS-overexpressing colorectal cancer

Modulating Tankyrases(TNKS),interactions with USP25 to promote TNKS degradation,rather than inhibiting their enzymatic activities,is emerging as an alternative/specific approach to inhibit the Wnt/β-catenin pathway.Here,we identified UAT-B,a novel neoantimycin analog isolated from Streptomyces conglobatus,as a small-molecule inhibitor of TNKS-USP25 protein-protein interaction(PPI)to overcome multi-drug resistance in colorectal cancer(CRC).The disruption of TNKS-USP25 complex formation by UAT-B led to a significant decrease in TNKS levels,triggering cell apoptosis through modulation of the Wnt/β-catenin pathway.Importantly,UAT-B successfully inhibited the CRC cells growth that harbored high TNKS levels,as demonstrated in various in vitro and in vivo studies utilizing cell line-based and patient-derived xenografts,as well as APC^(min/+)spontaneous CRC models.Collectively,these findings suggest that targeting the TNKS-USP25 PPI using a small-molecule inhibitor represents a compelling therapeutic strategy for CRC treatment,and UAT-B emerges as a promising candidate for further preclinical and clinical investigations.

Identification of an Aux/IAA regulator for flesh firmness using combined GWAS and bulked segregant RNA-Seq analysis in watermelon

Watermelon is a highly cultivated fruit crop renowned for its quality properties of fruit flesh.Among various quality factors,fruit flesh firmness is a crucial quality parameter influencing the fruit texture,shelf life and its commercial value.The auxin/indole-3-acetic acid(Aux/IAA)plays a significant role in fruit development and ripening of non-climacteric fruits.However,the regulatory mechanism of Aux/IAA in controlling fruit flesh firmness and ripening in watermelon remains unknown.In this study,we employed an integrative approach combining genome-wide association study(GWAS)and bulked segregant RNA-Seq analysis(BSR-Seq)to identify an overlapping candidate region between 12776310 and 12968331 bp on chromosome 6,underlying an auxin-responsive gene(Aux/IAA)associated with flesh firmness in watermelon.Transcriptome analysis,followed by real-time quantitative reverse transcription PCR(qRT-PCR),confirmed that the expression of Aux/IAA was consistently higher in fruits with high flesh firmness.The sequence alignment revealed a single base mutation in the coding region of Aux/IAA.Furthermore,the concomitant Kompetitive/Competitive allele-specific PCR(KASP)genotyping data sets for F2 population and germplasm accessions identified Aux/IAA as a strong candidate gene associated with flesh firmness.Aux/IAA was enriched in the plant hormone signal transduction pathway,involving cell enlargement and leading to low flesh firmness.We determined the higher accumulation of abscisic acid(ABA)in fruits with low flesh firmness than hard flesh.Moreover,overexpression of Aux/IAA induced higher flesh firmness with an increased number of fruit flesh cells while reducing ABA content and flesh cell sizes.Additionally,the allelic variation in Aux/IAA for soft flesh firmness was found to exist in Citrullus mucosospermus and gradually fixed into Citrullus lanatus during domestication,indicating that soft flesh firmness was a domesticated trait.These findings significantly enhanced our understanding of watermelon fruit flesh firmness and consequently the watermelon fruit quality.

Enhancer of Shoot Regeneration 2(ESR2)regulates pollen maturation and vitality in watermelon(Citrullus lanatus)

Watermelon(Citrullus lanatus)holds global significance as a fruit with high economic and nutritional value.Exploring the regulatory network of watermelon male reproductive development is crucial for developing male sterile materials and facilitating cross-breeding.Despite its importance,there is a lack of research on the regulation mechanism of male reproductive development in watermelon.In this study,we identified that ClESR2,a VIIIb subclass member in the APETALA2/Ethylene Responsive Factor(AP2/ERF)superfamily,was a key factor in pollen development.RNA insitu hybridization confirmed significantClESR2 expression in the tapetum and pollen during the later stage of anther development.The pollens of transgenic plants showed major defects in morphology and vitality at the late development stage.The RNA-seq and protein interaction assay confirmed that ClESR2 regulates pollen morphology and fertility by interacting with key genes involved in pollen development at both transcriptional and protein levels.These suggest that Enhancer of Shoot Regeneration 2(ESR2)plays an important role in pollen maturation and vitality.This study helps understand the male reproductive development of watermelon,providing a theoretical foundation for developing male sterile materials.

Transcription factor CabHLH035 promotes cold resistance and homeostasis of reactive oxygen species in pepper

Plant basic helix-loop-helix(bHLH)transcription factors(TFs)play central roles in various abiotic stresses.However,its role in plant cold resistance is largely unknown.Previously,we characterised CaNAC035 in pepper,which positively regulates tolerance to cold,salt and drought stresses tolerance.Here,we identified CabHLH035,a CaNAC035-interacting protein in pepper.To explore its functions in cold stress tolerance,we silenced the gene in pepper via virus-induced gene silencing(VIGS)and overexpressed the gene in Arabidopsis.The results showed that CabHLH035 expression was induced by cold treatment,and silencing of CabHLH035 decreased cold stress tolerance.Conversely,overexpression of CabHLH035 in Arabidopsis increased cold stress tolerance.To investigate homologs genes of C-repeat binding factor(CBF)pathway proteins and reactive oxygen species(ROS)marker gene expression blocking by CabHLH035,we performed yeast one-hybrid(Y1H),dual luciferase and electrophoretic mobility shift assay experiments.The results showed that CabHLH035 bound to the region upstream of the CaCBF1A and CaAPX promoters.Additionally,CaCBF1A bound to the CaDHN4 promoter.Taken together,our results showed that CabHLH035 plays a crucial role in cold stress tolerance and its potential as a target for breeding cold-resistant crops.The findings provide a basis for studying the functions and regulatory network of cold stress tolerance in pepper.

Chemokine-like factor 1 (CKLF1) is expressed in myocardial ischemia injury in vivo and in vitro

Introduction:Chemokine-like factor 1(CKLF1)is a chemokine that is overexpressed in several diseases.Our previousfindings revealed a significant increase in CKLF1 expression in the ischemic brain,suggesting its potential as a therapeutic target for ischemic stroke.Methods:In this study,we examined the expression dynamics of CKLF1 in both in vivo and in vitro models of ischemic cardiac injury.Myocardial infarction(MI)was induced in vivo by ligation of the left anterior descending artery(LAD)of the rat heart.The levels of CKLF1,Creatine Kinase MB Isoenzyme(CK-MB),and Lactate dehydrogenase(LDH)in the serum were detected using Enzyme-linked immunosorbent assay(ELISA).The expression of CKLF1 in the infarcted area was detected by immunohistochemistry,immunofluorescence,quantitative PCR(qPCR),and Western blotting(WB).H9C2 and AC16 cardiomyocytes cultured in vitro were subjected to oxygen and glucose deprivation(OGD).LDH was used to detect cell damage,and CKLF1 expression was detected by qPCR and WB.Results:CKLF1 mRNA and protein expression were significantly increased in h9c2 cells at 1.5 h and in AC16 cells at 4 h after OGD.The serum CK-MB in rats increased significantly on thefirst day after infarction,while the LDH concentration increased significantly on the third day after infarction.CKLF1 blood levels significantly increased on thefirst day following MI in rats.CKLF1 expression notably increased in the infarct area on days 1,3,and 7 post-MI.In MI tissue,CKLF1 colocalizes with cardiomyocytes,macrophages,and neutrophils.Conclusion:CKLF1 was substantially expressed during myocardial ischemia injury both in vivo and in vitro and was colocalized with macrophages and neutrophils,indicating that CKLF1 is expected to be a biomarker and a drug target for the treatment of myocardial infarction.

Therapeutic strategies targeting the epidermal growth factor receptor signaling pathway in metastatic colorectal cancer

More than 1.9 million new colorectal cancer(CRC)cases and 935000 deaths were estimated to occur worldwide in 2020,representing about one in ten cancer cases and deaths.Overall,colorectal ranks third in incidence,but second in mortality.More than half of the patients are in advanced stages at diagnosis.Treatment options are complex because of the heterogeneity of the patient population,including different molecular subtypes.Treatments have included conventional fluorouracil-based chemotherapy,targeted therapy,immunotherapy,etc.In recent years,with the development of genetic testing technology,more and more targeted drugs have been applied to the treatment of CRC,which has further prolonged the survival of metastatic CRC patients.

TM9SF1 is implicated in promoting the proliferation and invasion of bladder cancer cells

Zhuo et al looked into the part of transmembrane 9 superfamily member 1(TM9SF1)in bladder cancer(BC),and evaluated if it can be used as a therapeutic target.They created a permanent BC cell line and tested the effects of TM9SF1 overexpression and suppression on BC cell growth,movement,invasion,and cell cycle advancement.Their results show that TM9SF1 can boost the growth,movement,and invasion of BC cells and their access into the G2/M stage of the cell cycle.This research gives a novel direction and concept for targeted therapy of BC.

Recombinant Pichia pastoris overexpressing bioactive phytase

Phytase genephyA2, whose signal peptide encoding sequence and intron sequence had been removed, was modified. The Arg-encoding codons CGG and CAG inphyA2 were mutated into synonymous codon AGA. The modifiedphyA2 was fused behind a-factor signal sequence under the control ofAOX1 promoter in plasmid pPIC9, then introduced into the hostPichia pastoris by electroporation. The results of Southern blotting analysis and Northem blotting analysis demonstrated that thephyA2 gene had integrated into the genome ofP. pastoris and transcribed. The result of SDS-PAGE of the phytase expressed by P.pastoris showed that the modifiedphyA2 had been overexpressed and secreted. The concentration of the phytase expressed by P.pastoris with modifiedphyA2 exceeded 15 000 U/mL, which had a 3 000-fold increase over that of originAspergillus niger 963 and was 37 times higher than that of recombinantP. pastoris with non-modifiedphyA2.

The Improvement of Soybean Salt Tolerance by Overexpressed GmPAO1

Polyamines play an important regulatory role during plant growth and development and adversity stress,and polyamine oxidase(PAO)is involved in polyamine catabolism.In this study,an up-regulated polyamine oxidase gene GmPAO1 was obtained by transcriptome sequencing analysis and screening at soybean seedling stages.Also,its expression pattern and function were analyzed.The identification results of transgenic GmPAO1 soybean positive lines showed that the relative expression level of GmPAO1 in the overexpressed lines was increased under salt stress.With increasing stress concentration,the seed germination rate decreased.However,the seed germination rate of the overexpressed lines was significantly higher than that of the control lines,and the phenotypic character of the root systems was also better than that of the control lines.The measurement of superoxide dismutase(SOD)and peroxidase(POD)activities and malondialdehyde and hydrogen peroxide contents revealed that the overexpressed soybean lines significantly increased the SOD and POD activities,significantly reducing the malondialdehyde content.Although the hydrogen peroxide content in the transformed plants gradually increased,the hydrogen peroxide content in the overexpression lines was still lower than that in the gene editing lines.Based on this,it was preliminarily judged that GmPAO1 can improve soybean salt tolerance.

Comparative Non Participating Transcriptome Analysis Response to Low Phosphorus by CmPht1;2 in Chrysanthemum

Chrysanthemum morifolium Ramat is one of the four major cut flowers in the world.Pht1 family is focus on the uptake and transport of phosphate in plants.In our previous studies,CmPht1;2 overexpression line(Oe1)had higher phosphate contents both in roots and shoots,and its root development was significantly enhanced than wild type(WT)at low phosphorus conditions in chrysanthemum.Metabolomics analysis showed that several metabolites had a change in pyruvate metabolism and tricarboxylic acid(TCA)cycle pathway.To explore the gene difference expression and the change of metabolic pathway between CmPht1;2-Oe1 and WT,we conducted the transcriptome analysis.A total of 617,681 and 207,271 unigenes were obtained from roots and shoots,respectively.They were classified into biological process,cellular component and molecular function by Gene Ontology(GO).In addition,450 different expression genes(DEGs)were found in the roots after 2 d treatment,and 1,787 DEGs were identified in shoots after 7 d treatment under LP condition between Oe1 and WT.From the top 20 pathways of DEGs assigned by Kyoto Encyclopedia of Genes and Genomes(KEGG),TCA cycle and pyruvate metabolism pathways mostly affected by overexpression of CmPht1;2 attracted our attention.This research will be helpful for elucidating the mechanism of effects by CmPht1;2 overexpression on growth,development and stress tolerance.

GmSKP1,a Novel S-phase Kinase-associated Protein 1 in Glycine max,Enhancing Resistance Against Phytophthora sojae Infection

Phytophthora root and stem rot of soybean caused by Phytophthora sojae(P.sojae)is a devastating disease that affects soybean[Glycine max(L.)Merr.]all over the world.S-phase kinase-associated protein 1(SKP1)proteins are key members of the SKP1/Cullin/F-box protein(SCF)ubiquitin ligase complex and play diverse roles in plant biology.However,the role of SKP1 in soybean against the phytopathogenic oomycete P.sojae remains unclear.In this study,a novel member of the soybean SKP1 gene family,GmSKP1 which was significantly induced by P.sojae,was reported.The expression of GmSKP1 was simultaneously induced by methyl jasmonate(MeJA),salicylic acid(SA)and ethylene(ET),which might suggest an important role for GmSKP1 of plant in responses to hormone treatments.Functional analysis using GmSKP1 overexpression lines showed that GmSKP1 enhanced resistance to P.sojae in transgenic soybean plants.Further analyses showed that GmSKP1 interacted with a homeodomain-leucine zipper protein transcription factor(GmHDL56)and a WRKY transcription factor(GmWRKY31),which could positively regulate responses to P.sojae in soybean.Importantly,several pathogenesis-related(PR)genes were constitutively activated,including GmPR1a,GmPR2,GmPR3,GmPR4,GmPR5a and GmPR10,in GmSKP1-OE soybean plants.Taken together,these results suggested that GmSKP1 enhanced resistance to P.sojae in soybean,possibly by activating the defense-related PR genes.

Molecular Cloning, Escherichia coli Expression and Genomic Organization of Squalene Synthase Gene from Artemisia annua

A 1 539 by squalene synthase (AaSQS) cDNA was cloned from a high-yield Artemisia annua L. strain 001 by reverse transcription-polymerise chain reaction (RT-PCR). The amino acid sequence of AaSQS is 70%, 77%, 44% and 39%a identical to that of squalene synthases from Arabidopsis thaliana, tobacco, human and yeast, respectively. The AaSQS genomic DNA has a complex organization containing 14 exons and 13 introns. Full-length or C-terminal truncated cDNA was subcloned into prokaryotic expression vector pET30a and the constructed plasmid was introduced to Escherichia coli strain BL21 (DE3) for induced overexpression. No squalene synthase protein with expected molecular mass was observed in E. cola containing the putative full-length squalene synthase cDNA, however, overexpression in E. coli was achieved by truncating 30 amino acids of hydrophobic region at the carboxy terminus.

Increased Endogenous Methyl Jasmonate Altered Leaf and Root Development in Transgenic Soybean Plants

Methyl jasmonate （MeJA） is a plant-signaling molecule that regulates plant morphogenesis and expression of plant defense genes. To determine the role of the endogenous MeJA levels in the development of plants, transgenic soybean [Glycine max （L.） Merrill] plants harboring NTR1 gene encoding for jasmonic acid carboxyl methyltransferase （JMT） were produced. The activation of NTR1 gene expression resulted in the production of MeJA. Overexpression of the NTR1 cDNA under the regulation of cauliflower mosaic virus （CaMV） 35S promoter in the transgenic soybean plants was confirmed using Northern blot analysis. The significant differences in leaf and root growth patterns were observed between the transgenic plants and the wild-type plants. The leaves of the transgenic plants were slightly elongated in length but dramatically narrowed in width compared with the nontrans-formed wild-type plants. In addition, elongation of primary root was inhibited in the overexpressed transgenic soybean plantlets, whereas the development of lateral root was stimulated relative to the nontransformed plants. The leaves of the transgenic plants showed 2-2.5-fold higher levels of MeJA than the control plants. These results indicated that the increased endogenous levels of MeJA is involved in regulation of morphogenesis in soybean plants.

Effect of diacylglycerol acyltransferase 2overexpression in 3T3-L1 is associated to an increase in mono-unsaturated fatty acid accumulation

Background：Fatty acid（FA） composition is the most important parameter affecting the flavor and nutritional value of the meat.The final and the only committed step in the biosynthesis of triglycerides is catalyzed by diacylglycerol acyltransferase 2（DGAT2）.The role of DGAT2 in lipid accumulation has been demonstrated in adipocytes,However,little is known about the effect of DGAT2 on the FA composition of these cells.Methods：To investigate the role of DGAT2 in regulating lipid accumulation,FA composition and the expression of adipogenic genes,we cloned the open reading frame of the porcine DGAT2 gene and established 3T3-L1 cells that overexpressed DGAT2.Cells were then cultured in differentiation medium（DM） without FA,with a mixture of FAs（FA-DM）,or containing a ^（13）C stable isotope-labeled FA mixture（IFA-DM）.The FA composition of adipocytes was analyzed by gas chromatography-mass spectrometry and gas chromatography-isotope ratio mass spectrometry.Quantitative PCR and western blotting were employed to detect expression of adipogenic genes in 3T3-L1 adipocytes cultured with FA-DM for 12 d.Results：The triacylglyceride（TAG） content was significantly higher in 3T3-L1 adipocytes overexpressing DGAT2 than in control cells.When cultured in DM or FA-DM for 12 d,cells overexpressing DGAT2 showed a higher proportion of unsaturated FAs（C16：1 and C18：1）.However,when cells overexpressing DGAT2 were cultured with FA-DM for30 min,the FA composition was almost identical to that of controls.Further,the proportion of stable isotope-labeled FAs were similar in 3T3-L1 adipocytes overexpressing DGAT2 and control cells cultured in IFA-DM for 12 d.These results collectively indicate that the higher proportion of mono-unsaturated FAs,C16：1 and C18：1,may originate from de novo FA synthesis but not from the uptake of specific FAs from the medium.This hypothesis is further supported by evidence that both mRNA and protein expression of genes involved in FA synthesis（ACACA,FASN,SCD1,and A-FABP）were significantly higher in cells overexpressing DGAT2 than in control cells.Conclusions：In conclusion,our study revealed that TAG accumulation,the proportion of MUFAs,and the expression of adipogenic genes were higher in 3T3-L1 cells overexpressing DGAT2 than in control cells.

Overexpression of B7-H3 augments anti-apoptosis of colorectal cancer cells by Jak2-STAT3

AIM:To investigate the role of the overexpression of B7-H3 in apoptosis in colorectal cancer cell lines and the underlying molecular mechanisms.METHODS:SW620 cells that highly overexpressed B7-H3(SW620-B7-H3-EGFP)and HCT8 cells stably transfected with B7-H3 sh RNA(HCT8-sh B7-H3)were previously constructed in our laboratory.Cells transfected with p IRES2-EGFP were used as negative controls(SW620-NC and HCT8-NC).Real-time PCR and western blotting analysis were used to detect the m RNA and protein expressions of the apoptosis regulator proteins Bcl-2,Bcl-xl and Bax.A cell proliferation assay was used to evaluate the survival rate and drug sensitivity of the cells.The effect of drug resistance was detected by a cell cycle assay.Active caspase-3western blotting was used to reflect the anti-apoptotic ability of cells.Western blotting was also performed to determine the expression of proteins associated with the Jak2-STAT3 signaling pathway and the apoptosis regulator proteins after the treatment with AG490,a Jak2 specific inhibitor,in B7-H3 overexpressing cells.The data were analyzed by Graph Pad Prism 6 using a non-paired t-test.RESULTS:Whether by overexpression in SW620cells or downregulation in HCT8,B7-H3 significantly affected the expression of anti-and pro-apoptotic proteins,at both the transcriptional and translational levels,compared with the negative control(P<0.05).A cell proliferation assay revealed that B7-H3overexpression increased the drug resistance of cells and resulted in a higher survival rate(P<0.05).In addition,the results of cell cycle and active caspase-3western blotting proved that B7-H3 overexpression inhibited apoptosis in colorectal cancer cell lines(P<0.05).B7-H3 overexpression improved Jak2 and STAT3phosphorylation and,in turn,increased the expression of the downstream anti-apoptotic proteins B-cell CLL/lymphoma 2(Bcl-2)and Bcl-xl,based on western blotting(P<0.05).After treating B7-H3 overexpressing cells with the Jak2-specific inhibitor AG490,the phosphorylation of Jak2 and STAT3,and the expression of Bcl-2 and Bcl-xl,decreased accordingly(P<0.05).This finding suggested that the Jak2-STAT3 pathway is involved in the mechanism mediating the anti-apoptotic ability of B7-H3.CONCLUSION:The overexpression of B7-H3 induces resistance to apoptosis in colorectal cancer cell lines by upregulating the Jak2-STAT3 signaling pathway,potentially providing new approaches to the treatment of colorectal cancer.

Prognosis of HER2 over-expressing gastric cancer patients with liver metastasis

AIM:To study the risk factors for liver metastasis and the prognosis in patients with human epidermal growth factor receptor 2(HER2) over-expressing gastric cancer(GC).METHODS:A total of 84 GC patients recruited from the General Hospital of the People's Liberation Army(PLA) between 2003 and 2010 were randomly enrolled in this study.HER2 expression was detected by immunohistochemistry in 84 GC patients with liver metastases.The study group consisted of 66 men and 18 women,with an average age of 54 years(range:19-74 years).Liver metastasis was diagnosed by magnetic resonance imaging or computed tomography.Patients were followed-up and predictive factors of liver metastasis were evaluated.RESULTS:The median follow-up period was 47 mo(range:6-85 mo).The characteristics of 35(25.7%) patients with HER2 over-expression of liver metastatic GC are presented.HER2 over-expression was detected in 23 out of 49(46.9%) patients with intestinal GC,and 9 out of 35(25.7%) patients with diffuse GC.29 out of 59(49.2%) patients aged < 60 years were HER2positive,while 8 out of 25(32%) patients aged ≥ 60 were HER2-positive;a significant difference(P < 0.05).Univariate analysis(log-rank test) showed that HER2 over-expression,sex,Lauren classification,differentiation and disease-free interval were correlated with poor survival(P < 0.05).Survival analysis with a survival curve showed that HER2 over-expression was significantly relevant,with a reduced survival time in GC patients with liver metastases(P < 0.01).2-year survival was not associated with the patient's age.A diseasefree survival longer than 12 mo has a significant association with extended overall survival(OS) in GC patients with liver metastases.The median survival time after the diagnosis of liver metastases was 18 mo [95% confidence interval(CI):9.07-26.94] among HER2 positive GC patients with liver metastases.In comparison,for 49(69.4%) out of 84 HER2 negative patients with liver metastatic GC,the median survival time was 47 mo(95% CI:19.37-74.63).In patients with HER2 positive liver metastatic GC,the median OS was significantly shorter than in HER2 negative patients(median,20.32 mo;95% CI:16.51-24.13 vs median,50.14 mo;95% CI:37.83-62.45;P < 0.01).CONCLUSION:HER2 over-expressing GC patients with liver metastases have a poor prognosis.Overall survival was significantly lower in HER2 positive patients.HER2overexpression is correlated with a lower survival rate.