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Objective To investigate the role and mechanism of eukaryotic translation elongation factor 1(EEF1) family members (EEF1D,EEF1A1,and EEF1A2) in lung adenocarcinoma (LUAD) based on public databases.Methods We examined EEF1 member expression levels in human LUAD samples via The Cancer Genome Atlas in the UCSC Xena browser and the Clinical Proteomic Tumor Analysis Consortium.We analyzed the mRNA and protein levels of EEF1D,EEF1A1,and EEF1A2 and their correlations with pathological variables via the Mann-Whitney U test.The Kaplan-Meier curves were established to assess the prognostic values of EEF1D,EEF1A1,and EEF1A2.The single-sample gene set enrichment analysis algorithm was employed to explore the relationship between the expression levels of EEF1 members and tumor immune cell infiltration.Spearman and Pearson correlation analyses were performed to examine the relationship between the expression levels of EEF1 members and those of the genes in the phosphatidylinositol 3-kinase/protein kinase B signaling pathway.The immunohistochemical assay was employed to determine the expression levels of EEF1D,EEF1A1,and EEF1A2 in the LUAD tissue (n=75) and paracancer tissue (n=75) samples.Results The mRNA and protein levels of EEF1D,EEF1A1,and EEF1A2 showed significant differences between tumor and paracancer tissues (all P<0.001).The patients with high protein levels of EEF1A1 showed bad prognosis in terms of overall survival (P=0.039),and those with high protein levels of EEF1A2 showed good prognosis in terms of overall survival (P=0.012).The influence of the mRNA level of EEF1D on prognosis was associated with pathological characteristics.The expression levels of EEF1 members were significantly associated with the infiltration of various immune cells and the expression of key molecules in the phosphatidylinositol 3-kinase/protein kinase B signaling pathway.Conclusion EEF1D,EEF1A1,and EEF1A2 are associated with the progression of LUAD,serving as the candidate prognostic markers for LUAD.
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Humans , Peptide Elongation Factor 1/metabolism , Proteomics , Proto-Oncogene Proteins c-akt/metabolism , Carcinogenesis , Adenocarcinoma of Lung , Lung Neoplasms , RNA, Messenger/genetics , Phosphatidylinositol 3-Kinases , PrognosisABSTRACT
Aims@#Knowledge of the Trichoderma taxa is important for both control efficiency and environmental conservation. Therefore, the objective of this study is to isolate and identify Trichoderma species from various rhizosphere soil samples using phenotypic and molecular characterization.@*Methodology and results@#Native Trichoderma spp. were isolated from agricultural fields in 17 sites from seven states of Malaysia. These isolates were characterized via morphological observation and molecular phylogenetic analysis based on the translation elongation factor-1α (tef1-α) gene. About 42 isolates were classified into eight Trichoderma species, which are Trichoderma asperellum, T. hamatum, T. harzianum, T. koningiopsis, T. rodmanii, T. spirale, T. viride and T. virens. Comparison of DNA sequences of tef1-α showed that the isolates were 98-100% similar to respective Trichoderma species from GenBank, thus confirming the fungal identity. Phylogenetic trees of maximum likelihood (ML) dataset of tef1-α inferred that the isolates were clustered according to species.@*Conclusion, significance and impact of study@#Findings in the present study will be beneficial for the purposes of biodiversity conservation and plant disease management using biocontrol agents.
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RhizosphereABSTRACT
Objective: To study the effects of eukaryotic translation elongation factor 1A1 (eEF1A1) on the cell cycle and proliferation of hepatocellular carcinoma (HCC) cells, and to explore the molecular mechanism. Methods: The expression of eEF1A1 protein in 101 HCC tissues and their adjacent tissues were detected by immunohistochemistry, and the correlations between the expression of eEF1A1 protein and the clinicopathologic parameters of HCC patients was further analyzed. The expressions of eEF1A1 mRNA and protein in HCC cells (including Huh7, SMMC7721, MHCC97H and Hep3B) and the normal liver cells HL-7702 were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The specific shRNA-1/2 targeting eEF1A1 gene (eEF1A1-shRNA-1/2) and the overexpression plasmid pcDNA3.1- eEF1A1 were respectively transfected into Huh7 and SMMC7721 cells, and the alteration of eEF1A1 expression was verified by real-time fluorescent quantitative PCR and Western blotting, respectively. Then the proliferation and cell cycle of HCC cells were detected by EdU and FCM assays, respectively. In addition, the expressions of cyclin D2 and the signal transducer and activator of transcription 1 (STAT1)-related pathway proteins in Huh7 cells transfected with eEF1A1-shRNA-1/2 and in SMMC7721 cells transfected with pcDNA3.1- eEF1A1 were detected by Western blotting. Results: The expression level of eEF1A1 protein in HCC tissues was markedly higher than that in the adjacent tissues (P 0.05) in SMMC7721 cells after eEF1A1 overexpression and treatment with STAT1 inhibitor fludarabine. Conclusion: eEF1A1 regulates the expression of cyclin D2 through STAT1 pathway, thereby promoting the cell cycle progression and proliferation of primary HCC cells.
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<p><b>OBJECTIVE</b>To explore the role of eukaryotic translation elongation factor 1A1 (eEF1A1) in regulating the invasion and metastasis of hepatocellular carcinoma (HCC) cells and the possible mechanism.</p><p><b>METHODS</b>qRT-PCR and Western blotting were used to detect the mRNA and protein expression of eEF1A1 and NOB1 in different HCC cell lines and normal liver cells. The invasion and migration abilities of HCC cells with eEF1A1 knockdown or overexpression were examined using Transwell chamber assay and RTCA assay, and the changes in NOB1 mRNA and protein expressions in the cells were detected. The effects of increasing NOB1 expression in HCCLM3-sheEF1A1 cells and decreasing NOB1 expression in eEF1A1-overexpressing MHCC97h cells on eEF1A1 expression and cell invasion and migration abilities were analyzed using Western blotting, Transwell chamber assay and RTCA assay.</p><p><b>RESULTS</b>The expressions of eEF1A1 and NOB1 were significantly increased in positive correlation in HCC cells as compared with normal hepatocytes. Knockdown of eEF1A1 significantly decreased the invasion and migration of HCC cells and reduced the mRNA and protein expression of NOB1 ( < 0.01). Overexpression of eEF1A1 significantly enhanced invasion and migration of HCC cells and increased NOB1 mRNA and protein expressions ( < 0.01). Increasing NOB1 expression in HCCLM3-sheEF1A1 cells led to the restoration of NOB1 expression and cell invasion and migration abilities ( < 0.01), whereas decreasing NOB1 in MHCC97h-eEF1A1 cells resulted in inhibition of NOB1 expression and cell invasion and migration ( < 0.01).</p><p><b>CONCLUSIONS</b>eEF1A1 positively regulates the expression of NOB1 to promote the invasion and migration of HCC cells .</p>
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BACKGROUND: The prevalence of novel type 1 diabetes mellitus (T1DM) antibodies targeting eukaryote translation elongation factor 1 alpha 1 autoantibody (EEF1A1-AAb) and ubiquitin-conjugating enzyme 2L3 autoantibody (UBE2L3-AAb) has been shown to be negatively correlated with age in T1DM subjects. Therefore, we aimed to investigate whether age affects the levels of these two antibodies in nondiabetic subjects. METHODS: EEF1A1-AAb and UBE2L3-AAb levels in nondiabetic control subjects (n=150) and T1DM subjects (n=101) in various ranges of age (18 to 69 years) were measured using an enzyme-linked immunosorbent assay. The cutoff point for the presence of each autoantibody was determined based on control subjects using the formula: [mean absorbance+3×standard deviation]. RESULTS: In nondiabetic subjects, there were no significant correlations between age and EEF1A1-AAb and UBE2L3-AAb levels. However, there was wide variation in EEF1A1-AAb and UBE2L3-AAb levels among control subjects <40 years old; the prevalence of both EEF1A1-AAb and UBE2L3-AAb in these subjects was 4.4%. When using cutoff points determined from the control subjects <40 years old, the prevalence of both autoantibodies in T1DM subjects was decreased (EEFA1-AAb, 15.8% to 8.9%; UBE2L3-AAb, 10.9% to 7.9%) when compared to the prevalence using the cutoff derived from the totals for control subjects. CONCLUSION: There was no association between age and EEF1A1-AAb or UBE2L3-AAb levels in nondiabetic subjects. However, the wide variation in EEF1A1-AAb and UBE2L3-AAb levels apparent among the control subjects <40 years old should be taken into consideration when determining the cutoff reference range for the diagnosis of T1DM.
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Humans , Young Adult , Antibodies , Autoantibodies , Diabetes Mellitus, Type 1 , Diagnosis , Enzyme-Linked Immunosorbent Assay , Eukaryota , Peptide Elongation Factor 1 , Peptide Elongation Factors , Prevalence , Reference ValuesABSTRACT
Acanthopanax divaricatus, a member of the Araliaceae family, has been used as an invigorant in traditional Korean medicine. During disease monitoring, a stem with small, irregular, brown lesions was sampled at a farm in Cheonan in 2011. The symptoms seen were sunken cankers and reddish-brown needles on the infected twig. The isolated fungal colonies were whitish, having crenated edges and aerial mycelium on the surface, and with black gregarious fruiting bodies. The reverse plate was creamy white. Conidia were 17~22 x 3.5~4.2 microm, fusiform, 4-septate, and straight to slightly curved. The nucleotide sequence of the partial translation elongation factor 1 alpha gene of the fungal isolate, shares 99% sequence identity with that of known Pestalotiopsis ellipsospora. Based on the results of the morphological and molecular analyses, the fungal isolate was identified as P. ellipsospora. In Korea, this is the first report of canker on A. divaricatus.
Subject(s)
Humans , Eleutherococcus , Araliaceae , Base Sequence , Fruit , Korea , Mycelium , Needles , Peptide Elongation Factor 1 , Spores, FungalABSTRACT
Objective: To investigate the effects of RNA interference targeting eukaryotic translation elongation factor 1A1(eEF1A1) expression on the proliferation of hepatocellular carcinoma Hep3B cells, and to explore its possible mechanism. Methods: A recombinant vector pGPU6/GFP/Neo-eEF1A1-shRNA containing short hairpin RNA (shRNA) targeting eEF1A1 was constructed, and then it was transfected into the Hep3B cells. After tansfection with pGPU6/GFP/Neo-eEF1A1-shRNA, the expressions of eEF1A1mRNA and protein were examined by real time fluorescence quantitative PCR and Western blotting, respectively, the cellular growth ability was examined by cell counting kit 8 (CCK-8) assay, the colony formation ability was detected by colony formation assay, and the cell cycle distribution of Hep3B cells was detected by flow cytometry (FCM). The expression levels of cyclin D1 and cyclin-dependent kinase 4 (CDK4) proteins in Hep3B cells were examined by Western blotting. Results: The recombinant vector pGPU6/GFP/Neo-eEF1A1-shRNA was successfully constructed, and the Hep3B cells with stable expession of eEF1A1 were established. The expression levels of eEF1A1mRNA and protein in Hep3B cells after transfection with pGPU6/GFP/NeoeEF1A1-shRNA were lower than those in the negative control cells (Hep3B cells transfected with a negative control vector pGPU6/GFP/Neo-NC) and the blank control cells (Hep3B cells without any transfection) (P < 0.01, P < 0.05). As compared with the negative control, the cellular growth ability and the colony formation ability of Hep3B cells after tranfection with pGPU6/GFP/Neo-eEF1A1-shRNA were obviously decreased (P < 0.05, P < 0.01); the percentage of the cells in G1 phase was increased (P < 0.01), and which in S-phase was decreased (P < 0.05); the expression levels of cyclin D1and CDK4 proteins were down-regulated (both P < 0.05). Conclusion: Down-regulation of eEF1A1 expression can inhibit the proliferation of Hep3B cells.
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Objective To investigate the effects of over-expression of homo sapiens eukaryotic translation elongation factor 1 alpha 2 (EEF1A2) on in vitro invasion and lung metastasis of human pancreatic cancer SW1990 cells.Methods Letivirus-mediated delivery of EEF1A2 was used to enhance the expression of EEF1A2 gene in human pancreatic cancer SW1990 cells,the SW1990 cells stably over-expressing EEF1A2 protein (SW1990/EEF1A2 cells) were obtained,and the parent SW1990 cells and SW1990/GFP cells were used as the control,and the expressions of EEF1 A2 mRNA and protein were determined by Real-time PCR and Western blotting.The invasion ability of cells was determined by Transwell assay.The lung metastasis model was established by injection of SW1990 cells into the tail vein.Whole lung tissues were harvested,and visible nodules on tung surface were counted macroscopically 8 weeks later.Results The EEF1 A2 mRNA expression of SW1990/EEF1A2 was 3.252 ± 0.344,which was significantly higher than those in SW1990/GFP cells (1.000 ±0.060) and SW1990 cells (0.944 ±0.041,t =2.255,2.305,P＜0.01) ; the EEF1A2 protein expression was 0.833 ± 0.050,which was significantly higher than those in SW1990/GFP cells (0.247 ± 0.035) and SW1990 cells (0.273± 0.041,t=0.572,0.559,P＜0.01).The ability of invasion of SW1990/EEF1A2 cells was (60 ±4) cells,which was sigmificantly higher than (33 ±4) cells in SW1990/GFP group and (26 ± 3) cells in SW1990 group (t =31.33,34.78,P ＜ 0.01).Furthernore,SW1990/EEF1 A2 cells had a much higher incidence of lung metastasis in nude mice than SW1990/GFP cells and SW1990 cells in vivo (100％ vs.20％,20％,P ＜ 0.05).Conclusions EEF1 A2 over-expression can obviously increase the in vitro invasion and lung metastasis of pancreatic cancer SW1990 cells.
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Objective: To investigate the effect of targeted gene silencing of homo sapiens EEF1A2 (eukaryotic translation elongation factor 1 alpha 2) on migration and invasion of human pancreatic cancer cell line BxPC-3 in vitro , and to explore its possible molecular mechanism. Methods: The mRNA and protein expression levels of EEF1A2 in four strains of human pancreatic cancer cells were detected by RT-PCR and Western blotting, respectively. EEF1A2-siRNA (small interference RNA) and negative control siRNA were transfected into BxPC-3 cells, respectively. Then the mRNA and protein expressions of EEF1A2 were determined by RT-PCR and Western blotting, respectively. The abilities of migration and invasion of BxPC-3 cells were determined by wound healing assay and Transwell invasion assay, respectively. The changes of expressions of p-Akt and total Akt proteins were detected by Western blotting. Results: Of the four strains of human pancreatic cancer cells, SW1990 cells displayed a low level of EEF1A2, and the remaining three strains of human pancreatic cancer cells including BxPC-3, Patu8988 and Panc-1 all displayed high expressions of EEF1A2. At 48 h post-transfection, the expression levels of EEF1A2 mRNA and protein in BxPC-3 cells transfected with EEF1A2-siRNA were significantly decreased as compared with those transfected with negative control siRNA or without any transfection (P 0.05). Conclusion: Targeted silencing of EEF 1A 2 by siRNA can obviously inhibit the migration and invasion abilities of human pancreatic cancer BxPC-3 cells in vitro . This effect may be associated with Akt signaling pathway regulated by EEF1A2. Copyright © 2012 by TUMOR.
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Objective To elucidate whether down-regulation of homo sapiens eukaryotic translation elongation factor 1 alpha 2 (EEF1A2) expression induces apoptosis in pancreatic cancer cells and its possible mechanisms. Methods Two siRNAs targeting human EEF1A2 were synthesized and the siRNA/liposome complexes were transfected into the pancreatic cancer cell line BxPC-3. RTPCR and Western blot were used to analyze the change of EEF1A2 expression and the apoptosis rate of BxPC-3 cells was studied using Annexin-V/PI assay. To identify the mechanisms involved, the apoptosis associated proteins such as caspase-3, caspase-8, caspase-9, PARP, cytochrome C and Bid were detected by Western blotting. Results Both EEF1A2-targeting siRNAs reduced the EEF1A2expression, and the No. 2 siRNA inhibited EEF1A2 expression to less than 25 % in mRNA and protein levels. Down-regulation of EEF1A2 expression in BxPC-3 cells enhanced cell apoptosis (15.28% ±3.65%) at a greater level than negative siRNA-expressing cells (10. 11% ± 3. 05%) or mock cells (9.41 % ±4.14 %). Furthermore, reduction of EEF1A2 activated the pro-caspase-8, pro-caspase-3,pro-caspase-9,PARP and Bid to their active forms, and increased the expression of cytochrome C.Conclusions These data suggest that EEF1A2 down-regulation could significantly induce apoptosis of pancreatic cancer cell line BxPC-3, which is likely mediated by the death receptor and mitochondrial apoptotic pathways.
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Although Fursarium oxysporum causes diseases in economically important plant hosts, identification of F. oxysporum formae speciales has been difficult due to confusing phenotypic classification systems. To resolve these complexity, we evaluated genetic relationship of nine formae speciales of F. oxysporum with random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), and translation elongation factor-1 alpha (EF-1alpha) gene. In addition, the correlation between mycotoxin content of fusaric acid and isolates based on molecular marker data was evaluated using the modified Mantel's test. According to these result, these fusaric acid-producing strains could not identify clearly, and independent of geographic locations and host specificities. However, in the identification of F. oxysporum formae speciales, especially, AFLP analysis showed a higher discriminatory power than that of a the RAPD and EF-1alpha analyses, all three techniques were able to detect genetic variability among F. oxysporum formae speciales in this study.
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Amplified Fragment Length Polymorphism Analysis , Classification , DNA , Fusaric Acid , Fusarium , Geographic Locations , Host Specificity , Korea , Peptide Elongation Factor 1 , PlantsABSTRACT
Objective To investigate the expression changes of translation elongation factor 1?1 in anti-BPDE transformed and carcinoma 16 HBEs. Methods Suppression subtractive hybridization (SSH), bioinformatics and semi-quantitative RT-PCR were applied. The cDNA of anti-BPDE transformed and carcinoma 16 HBE cells were used as tester respectively, and the cDNA of normal 16 HBE was used as driver, the library of subtractive hybridization were profiled and inverted into TA cloning vector after two times of hybridization and two times of PCR. After the screening, sequencing and analysis of sequences, semi-quantitative RT-PCR was performed accompanying to the inner reference of ?-actin. Results 9 differentially expressed fragments were consistent with translation elongation factor 1?1 in different regions in Genbank, and the expressions were up regulated in BPDE transformed and carcinoma 16 HBE cells. Conclusion Translation elongation factor 1?1 may be related to the transforming effect and carcinogenesis of anti-BPDE.