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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 343 records · Page 19
All sourcesNASA NTRSDOE OSTI
DOE OSTI2017

miR-141-3p functions as a tumor suppressor modulating activating transcription factor 5 in glioma

Glioma is the most common malignant primary brain tumor which arises from the central nervous system. Our studies reported that an anti-apoptotic factor, activating transcription factor 5 (ATF5), is highly expressed in malignant glioma specimens and cell lines. Downregulation by dominant-negetive ATF5 could repress glioma cell proliferation and accelerate apoptosis. Here, we further investigate the upstream factor which regulates ATF5 expression. Bioinformatic analysis showed that ATF5 was a potential target of miR-141-3p. Luciferase reporter assay verified that miR-141-3p specifically targeted the ATF5 3′-UTR in glioma cells. Functional studied suggested that miR-141-3p overexpression inhibited proliferation and promoted apoptosis of glioma cells (U87MG and U251). Xenograft experiments proved the inhibition of miR-141-3p on glioma growth in vivo. Moreover, exogenous ATF5 without 3′-UTR restored the cell proliferation inhibition triggered by miR-141-3p. Taken together, we put forward that miR-141-3p is a new upstream target towards ATF5. It can serve as a crucial tumor suppressor in regulating the ATF5-regulated growth of malignant glioma. - Highlights: • ATF5 was highly expressed in gliomas and was essential for cell proliferation and apoptosis. • MiR-141-3p specifically targeted the ATF5 3′-UTR in glioma cells. • MiR-141-3p was significantly downregulated in glioma tissues and cell lines. • MiR-141-3p inhibited glioma growth through repressing ATF5 expression in vitro and in vivo. • Exogenous expression of ATF5 rescued cells from proliferation inhibition triggered by miR-141-3p.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

miR-217 suppresses proliferation and promotes apoptosis in cardiac myxoma by targeting Interleukin-6

Cardiac myxoma (CM) is a prevalent primary cardiac tumor. miR-217 plays a vital role in tumorigenesis of various cancers, however, its role and underlying molecular mechanism in human CM remain poorly understood. Here, we reported that the expression of miR-217 was downregulated in CM tissues and inversely correlated with the expression of Interleukin-6 (IL-6) mRNA. Gain-of-function analysis indicated that overexpression of miR-217 inhibited the proliferation and promoted the apoptosis of the primary CM cells. Bioinformatics analysis showed that IL-6 was a direct target gene of miR-217, which is confirmed by the dual luciferase assays. Moreover, downregulation of IL-6 by small interference RNA (siRNA) mimicked the tumor-suppressive effects of miR-217 in CM. Furthermore, rescue experiments pointed out that restoration of IL-6 expression abrogated the anti-proliferative and pro-apoptotic effect induced by miR-217 overexpression in CM cells. Taken together, we validated that miR-217 could act as a tumor suppressor in CM by directly targeting 3′UTR of IL-6 gene, indicating that manipulation of miR-217 may be a potential therapeutic strategy for CM patients. - Highlights: • Lower expression of miR-217 and higher expression of IL-6 mRNA was observed in CM tissues. • miR-217 overexpression suppressed proliferation and promoted apoptosis of primary CM cells. • miR-217 targeted IL-6 3′-UTR to inhibit its expression. • Reintroduction of IL-6 reversed the effect of miR-217 overexpression on CM cell proliferation and apoptosis.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

MiR-361 targets Yes-associated protein (YAP) mRNA to suppress cell proliferation in lung cancer

Yes-associated protein (YAP) contributes to the development of multiple tumors, but the post-transcription modulation of YAP remains unexplored. Here, we present a new regulatory microRNA of YAP, miR-361, which directly targets YAP to inhibit cell proliferation in lung cancer. We used bioinformatics to predict that miR-361 could target 3′-untranslated region (3′UTR) of YAP mRNA. Luciferase reporter gene assays demonstrated that miR-361 could decrease the luciferase activities of 3′UTR vector of YAP. Furthermore, YAP expression was obviously abated by miR-361 using RT-PCR and immunoblotting in lung cancer A549 cells. In terms of function, MTT and colony formation analysis showed that ectopic miR-361 expression significantly suppressed cell proliferation in lung cancer. Notably, overexpressed YAP accelerated miR-361-bated proliferation of lung cancer cells. MiR-361 inhibitor promoted cell proliferation in lung cancer, but YAP RNA interference reversed miR-361 inhibitor-driven cell proliferation. Interestingly, miR-361 was capable of affecting the cell cycle in lung cancer progression. Finally, the negative correlation of miR-361 with YAP was found in clinical human lung cancer tissues. In conclusion, miR-361 targets 3′UTR of YAP mRNA to depress the proliferation of lung cancer cells. - Highlights: • MiR-361 is able to target 3′UTR of YAP mRNA. • MiR-361 suppresses the proliferation of lung cancer by inhibiting YAP. • YAP is negatively correlated with miR-361 in lung cancer tissues. • YAP is up-regulated in clinical lung cancer tissues.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

MiR-362-3p inhibits the proliferation and migration of vascular smooth muscle cells in atherosclerosis by targeting ADAMTS1

Atherosclerosis is a progressive condition of the large arteries that can cause coronary artery disease (CAD). Growing amounts of evidence have indicated that microRNAs (miRNAs, miRs) can be used as diagnostic biomarkers in many cellular processes associated with CAD. MiR-362-3p has been implicated in many biological cellular functions. However, little is known about the role of miR-362-3p during atherosclerosis. In the present study, significant downregulation of miR-362-3p was observed in 110 atherosclerotic CAD patients and not in the 84 controls. The upregulation of miR-362-3p was demonstrated to inhibit vascular smooth muscle cell (VSMC) proliferation and migration, and impede the G1/S cell cycle transition. Bioinformatics analysis indicated that a disintegrin and metalloproteinase with thrombospondin motifs 1 (ADAMTS1) was a direct target of miR-362-3p. Subsequent experiments demonstrated that miR-362-3p binds to the 3′-untranslated region (UTR) of ADAMTS1 and decreases its levels of mRNA and protein expression. Overexpression of ADAMTS1 partially restored the miR-362-3p-mediated inhibition of VSMC proliferation, cell cycle, and migration. Upregulation of ADAMTS1 in plasma samples was detected in atherosclerotic CAD patients. Taken together, our findings suggested that miR-362-3p inhibits the proliferation and migration of VSMCs by directly targeting ADAMTS1, which might provide novel insight into the molecular mechanisms underlying the action of miR-362-3p in atherosclerosis. - Highlights: • MiR-362-3p expression is downregulated in the plasma samples of atherosclerotic CAD patients. • MiR-362-3p overexpression inhibits VSMC proliferation, cell cycle progression, and migration. • ADAMTS1 is a direct target of miR-362-3p. • Dysfunction of miR-362-3p may contribute to the pathophysiology of atherosclerosis.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

miR-19 promotes osteosarcoma progression by targeting SOCS6

Highlights: • miR-19 was upregulated and SOCS6 was downregulated in OS. • miR-19 dramatically suppressed OS growth in vitro and in vivo. • SOCS6 is a direct target gene of miR-19 in OS. • miR-19 regulated SOCS6/JAK2/STAT3 pathway in OS. microRNAs (miRNAs) play critical roles in cancer development and progression. This study investigated the effects of miR-19 in human osteosarcoma (OS) development. Here, we showed that miR-19 was frequently upregulated in OS tissues and cell lines. Moreover the expression of miR-19 was associated with TNM stage, metastasis, size and poor overall survival. Mechanistically, miR-19 dramatically suppressed OS growth in vitro and in vivo. Bioinformatics analyses predicted that SOCS6 is a potential target gene of miR-19 in OS, which was confirmed by luciferase-reporter assay. We also found that SOCS6 expression was downregulated and negatively correlated with miR-19 expression in OS tissues clinically. Moreover, ectopic SOCS6 could reverse miR-19 induced OS growth. Finally, JAK2/STAT3 signaling pathway involves miR-19/SOCS6-mediated OS progression. Together, our data provide important evidence for miR-19 mediated SOCS6 in OS growth and revealed miR-19/SOCS6/JAK2/STAT3 pathway as a potential therapeutic strategy for OS patients.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

Comparative analysis of methicillin-sensitive and resistant Staphylococcus aureus exposed to emodin based on proteomic profiling

Highlights: • Emodin is effective against methicillin-sensitive and resistant Staphylococcus aureus strains. • Global proteomic profiling revealed 145 and 122 significant changes in MSSA and MRSA after emodin treatment, respectively. • Profiling interaction network analysis demonstrated emodin potentially targeted MRSA via multiple mechanisms. Emodin has a strong antibacterial activity, including methicillin-resistant Staphylococcus aureus (MRSA). However, the mechanism by which emodin induces growth inhibition against MRSA remains unclear. In this study, the isobaric tags for relative and absolute quantitation (iTRAQ) proteomics approach was used to investigate the modes of action of emodin on a MRSA isolate and methicillin-sensitive S. aureus ATCC29213(MSSA). Proteomic analysis showed that expression levels of 145 and 122 proteins were changed significantly in MRSA and MSSA, respectively, after emodin treatment. Comparative analysis of the functions of differentially expressed proteins between the two strains was performed via bioinformatics tools blast2go and STRING database. Proteins related to pyruvate pathway imbalance induction, protein synthesis inhibition, and DNA synthesis suppression were found in both methicillin-sensitive and resistant strains. Moreover, Interference proteins related to membrane damage mechanism were also observed in MRSA. Our findings indicate that emodin is a potential antibacterial agent targeting MRSA via multiple mechanisms.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

Quantitative proteome analysis of bovine mammary gland reveals protein dynamic changes involved in peak and late lactation stages

Highlights: • Revealed dynamic protein changes between two lactation stages in bovine mammary by TMT-based proteomics (first time). • Longitudinal design avoided the influence of different genetic backgrounds on the DEP detection. • 179 differentially expressed proteins (DEPs) identified. • Identified novel DEPs: AACS, DHCR7, GSTM3, SFRP1 and SFRP4. • Major pathways: PI3K-Akt, localization, lipid metabolic, amino acid transport immune system and apoptotic signaling. Mammary gland is an important organ for milk synthesis and secretion. It undergoes dramatic physiological changes to adapt the shift from peak to late lactation stage. Protein plays a final very vital role in many life functions, and the protein changes during different lactation stages potentially reflect the biology of lactation and the functions of mammary gland in cows. In current study, we adopted tandem mass tags label-based quantitative analysis technique and to investigate proteome changes occurring in bovine mammary gland from peak to late lactation stages. A total of 3753 proteins from mammary tissues taken at two lactation points from four individual cows by biopsy were quantified, out of which 179 proteins were expressed differentially between two stages. We observed five new DEPs (AACS, DHCR7, GSTM3, SFRP1 and SFRP4) and nine functional well-studies known proteins (PLIN2, LPIN1, PLIN3, GSN, CD74, MMP2, SOD1, SOD3 and GPX3) related to milk performance and mammary morphology. Bioinformatics analyses of the DEPs showed a majority of the up-regulated proteins during late lactation stage were related to apoptosis and immune process, while the downregulated proteins were mainly involved in localization, lipid metabolic and transport process. This suggests that the mammary gland can adapt to different molecular functions according to the biological need of the animal. From the integrated analysis of the differentially expressed proteins with known quantitative trait loci and genome-wide association study data, we identified 95 proteins may potentially affect milking performance. We expect findings in this study could be a valuable resource for future studies investigating the bovine proteome and functional studies.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

Characterization of the zinc-induced Shank3 interactome of mouse synaptosome

Highlights: • The synaptosomal Shank3 complexes incubated with different ZnCl{sub 2} concentrations were analyzed by mass spectrometry. • The 71 proteins that specifically interacted with Shank3 under extra ZnCl{sub 2} conditions were identified. • The zinc-induced Shank3 interactome is associated with mitochondria and brain disorders. Variants of the SHANK3 gene, which encodes a core scaffold protein of the postsynaptic density of excitatory synapses, have been causally associated with numerous brain disorders. Shank3 proteins directly bind zinc ions through their C-terminal sterile α motif domain, which enhances the multimerization and synaptic localization of Shank3, to regulate excitatory synaptic strength. However, no studies have explored whether zinc affects the protein interactions of Shank3, which might contribute to the synaptic changes observed after zinc application. To examine this, we first purified Shank3 protein complexes from mouse brain synaptosomal lysates that were incubated with different concentrations of ZnCl{sub 2}, and analyzed them with mass spectrometry. We used strict criteria to identify 71 proteins that specifically interacted with Shank3 when extra ZnCl{sub 2} was added to the lysate. To characterize the zinc-induced Shank3 interactome, we performed various bioinformatic analyses that revealed significant associations of the interactome with subcellular compartments, including mitochondria, and brain disorders, such as bipolar disorder and schizophrenia. Together, our results showing that zinc affected the Shank3 protein interactions of in vitro mouse synaptosomes provided an additional link between zinc and core synaptic proteins that have been implicated in multiple brain disorders.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Coatomer subunit beta 2 (COPB2), identified by label-free quantitative proteomics, regulates cell proliferation and apoptosis in human prostate carcinoma cells

Highlights: • Our paper offered an available method to identified and confirm some differential expression of genes in prostate cancer. • Our paper provided us a common method to research the function of gene for prostate cancer. • Our paper helps us to identify a novel cancer related gene to explain for development of novel targeted drugs. Label-free quantitative proteomics has broad applications in the identification of differentially expressed proteins. Here, we applied this method to identify differentially expressed proteins (such as coatomer subunit beta 2 [COPB2]) and evaluated the functions and molecular mechanisms of these proteins in prostate cancer (PCA) cell proliferation. Proteins extracted from surgically resected PCA tissues and adjacent tissues of 3 patients were analyzed by label-free quantitative proteomics. The target protein was confirmed by bioinformatics and GEO dataset analyses. To investigate the role of the target protein in PCA, we used lentivirus-mediated small-interfering RNA (siRNA) to knockdown protein expression in the prostate carcinoma cell line, CWR22RV1 cells and assessed gene and protein expression by reverse transcription quantitative polymerase chain reaction and western blotting. CCK8 and colony formation assays were conducted to evaluate cell proliferation. Cell cycle distributions and apoptosis were assayed by flow cytometry. We selected the differentiation-related protein COPB2 as our target protein based on the results of label-free quantitative proteomics. High expression of COPB2 was found in PCA tissue and was related to poor overall survival based on a public dataset. Cell proliferation was significantly inhibited in COPB2-knockdown CWR22RV1 cells, as demonstrated by CCK8 and colony formation assays. Additionally, the apoptosis rate and percentage of cells in the G{sub 1} phase were increased in COPB2-knockdown cells compared with those in control cells. CDK2, CDK4, and cyclin D1 were downregulated, whereas p21 Waf1/Cip1 and p27 Kip1 were upregulated, affecting the cell cycle signaling pathway. COPB2 significantly promoted CWR22RV1 cell proliferation through the cell cycle signaling pathway. Thus, silencing of COPB2 may have therapeutic applications in PCA.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Biomarker identification for statin sensitivity of cancer cell lines

Highlights: • Graphical modeling is used to find a biomarker signature for statin sensitivity. • Predictions from the model were experimentally validated in four human cell lines. • Enrichment analysis and predicted combination therapies show relevance of signature. Statins are potent cholesterol reducing drugs that have been shown to reduce tumor cell proliferation in vitro and tumor growth in animal models. Moreover, retrospective human cohort studies demonstrated decreased cancer-specific mortality in patients taking statins. We previously implicated membrane E-cadherin expression as both a marker and mechanism for resistance to atorvastatin-mediated growth suppression of cancer cells; however, a transcriptome-profile-based biomarker signature for statin sensitivity has not yet been reported. Here, we utilized transcriptome data from fourteen NCI-60 cancer cell lines and their statin dose-response data to produce gene expression signatures that identify statin sensitive and resistant cell lines. We experimentally confirmed the validity of the identified biomarker signature in an independent set of cell lines and extended this signature to generate a proposed statin-sensitive subset of tumors listed in the TCGA database. Finally, we predicted drugs that would synergize with statins and found several predicted combination therapies to be experimentally confirmed. The combined bioinformatics-experimental approach described here can be used to generate an initial biomarker signature for anticancer drug therapy.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Inhibition of CIP2A attenuates tumor progression by inducing cell cycle arrest and promoting cellular senescence in hepatocellular carcinoma

Highlights: • High expression level of CIP2A was frequently observed in hepatocellular carcinoma (HCC). • CIP2A expression level was associated with unfavorable overall and recurrence-free survival in HCC patients. • Inhibition of CIP2A attenuated HCC progression via inducing cell cycle arrest and promoting cellular senescence. CIP2A is a recent identified oncogene that inhibits protein phosphatase 2A (PP2A) and stabilizes c-Myc in cancer cells. To investigate the potential oncogenic role and prognostic value of CIP2A, we comprehensively analyzed the CIP2A expression levels in pan-cancer and observed high expression level of CIP2A in majority cancer types, including hepatocellular carcinoma (HCC). Based on a validation cohort including 60 HCC and 20 non-tumorous tissue samples, we further confirmed the high mRNA and protein expression levels of CIP2A in HCC, and found high CIP2A mRNA expression level was associated with unfavorable overall and recurrence-free survival in patients with HCC. Mechanistic investigations revealed that inhibition of CIP2A significantly attenuated cellular proliferation in vitro and tumourigenicity in vivo. Bioinformatic analysis suggested that CIP2A might be involved in regulating cell cycle. Our experimental data further confirmed CIP2A knockdown induced cell cycle arrest at G1 phase. We found accumulated cellular senescence in HCC cells with CIP2A knockdown, companying expression changes of senescence associated proteins (p21, CDK2, CDK4, cyclin D1, MCM7 and FoxM1). Mechanistically, CIP2A knockdown repressed FoxM1 expression and induced FoxM1 dephosphorylation. Moreover, inhibition of PP2A by phosphatase inhibitor rescued the repression of FoxM1. Taken together, our results showed that CIP2A was highly expressed in HCC. Inhibition of CIP2A induced cell cycle arrest and promoted cellular senescence via repressing FoxM1 transcriptional activity, suggesting a potential anti-cancer target for patients with HCC.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

MiR-181a contributes gefitinib resistance in non-small cell lung cancer cells by targeting GAS7

Tyrosine kinase inhibitors (TKIs) exert potent therapeutic efficacy in non-small cell lung cancers (NSCLC) harboring epidermal growth factor receptor (EGFR) activating mutations. However, a major impediment for the effective treatment is the development of drug resistance. Some evidence supports a role for miRNAs in modulating NSCLC TKIs resistance. Here we show that miR-181a is significantly up-regulated in gefitinib-resistant cells compared with gefitinib-sensitive cells. Upregulation of miR-181a caused resistance of gefitinib, whereas downregulation of miR-181a sensitized NSCLC cells to gefitinib. Furthermore, the miR-181a plasma levels were significantly increased in acquired gefitinib resistant NSCLC patients compared with the plasma levels prior to gefitinib treatment in each patient. Bioinformatics analysis and luciferase reporter assay showed that growth arrest-specific 7 (GAS7) was a direct target gene of miR-181a. A significant inverse correlation between the expression of miR-181a and GAS7 was identified in NSCLC tissues. Downregulation of GAS7 expression could antagonize gefitinib re-sensitivity in PC9GR mediated by knockdown of miR-181a via AKT/ERK pathways and epithelial-to-mesenchymal transition markers. Additionally, GAS7 expression was downregulated in a large cohort of NSCLC patients, and a high mRNA level of GAS7 was associated with improved overall survival. Collectively, our findings provide a novel basis for using miR-181a/GAS7-based therapeutic strategies to reverse gefitinib resistance in NSCLC.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

MiR-7-5p functions as a tumor suppressor by targeting SOX18 in pancreatic ductal adenocarcinoma

Highlights: • MiR-7-5p is down-regulated in pancreatic cancer tissues. • Down-regulation of miR-7-5p indicates poor prognosis for PDAC patients. • MiR-7-5p targets SOX18 to inhibit pancreatic cancer growth in vivo and vitro. • MiR-7-5p inhibits PANC-1 cell migration and invasion by targeting SOX18. • SOX18 activates JAK2/STAT3 pathway by promoting gp130 transcription. Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal malignancies. Recently, many kinds of microRNAs (miRNAs) have been found to play a significant role in development of PDAC. However, there is no investigation about expression and function of miR-7-5p in PDAC. In this study, we found that miR-7-5p was down-regulated in PDAC tissues and its low expression level indicated a poor survival rate for PDAC patients. By bioinformatic analysis, we found that miR-7-5p targeted SOX18, and there was a negative correlation between them in PDAC tissues. Then luciferase reporter and western blot assays were used to verify the binding of miR-7-5p on SOX18 3′UTR. Cell function assays demonstrated that miR-7-5p inhibited proliferation, migration and invasion of PANC-1 cells by targeting SOX18. The nude mouse tumorigenicity assay further proved that miR-7-5p targeted SOX18 to inhibit pancreatic cancer growth in vivo. In order to further understand the mechanism, we applied a transcription factor prediction tool to explore the underlying targets that transcripted by SOX18, and the result indicated that SOX18 was a transcription factor for gp130 (a subunit of IL-6 receptor), and ChIP assays was performed to prove this prediction. Furthermore, we detected the suppression of gp130/JAK2/STAT3 signaling pathway after silencing SOX18 in PANC-1 cells, which demonstrated the transcriptional activation role of SOX18 on gp130. Thus, our present study revealed that miR-7-5p targeted SOX18 to inhibit gp130/JAK2/STAT3 signaling pathway to exert its suppressing role in PDAC.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

The role of Foxq1 in proliferation of human dental pulp stem cell

Highlights: • Foxq1 was found to be up-regulated in dental pulp stem cell after calcium hydroxide stimulation. • Overexpression of Foxq1 promote proliferation of dental pulp stem cell, while inhibition of Foxq1 repress its proliferation. • MiR-320b depression after calcium hydroxide stimulation ease its inhibition on Foxq1, thus promote Foxq1 expression. This study aimed to investigate the role for Foxq1 in proliferation activity regulation of dental pulp stem cells (DPSCs). Proliferation of DPSC was induced by calcium hydroxide, then expression alteration of Foxq1 was evaluated. Lentivirus was employed to manipulate Foxq1 level in DPSC, and proliferation activities were evaluated. To look into mechanism regulating Foxq1 level after calcium hydroxide stimulation, expressions of various microRNAs were evaluated, then bioinformatics study and dual-luciferase study were carried out to confirm targeting relationship between microRNA and Foxq1. The result of our study indicated that proliferation activities of DPSCs were enhanced after calcium hydroxide stimulation, during which expression of Foxq1 was also up-regulated. Cell viability and progression from G1 to S phase were both improved with overexpression of Foxq1, and microRNAs profiling study and dual-luciferase result suggested miR-320b contributed to the up-regulation of Foxq1 after calcium hydroxide stimulation. These results suggested that miR-320b mediated Foxq1 up-regulation promote proliferation of dental pulp stem cells.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Identification of aberrant circular RNA expression and its potential clinical value in primary great saphenous vein varicosities

Highlights: • This study identified 232 significantly aberrant circRNAs in primary great saphenous vein varicosities (PGSVVs). • A circRNA-miRNA-gene co-expression network was created. • Hsa{sub c}irc{sub 0}006427, hsa{sub c}irc{sub 0}089810 and hsa{sub c}irc{sub 0}005267 might be potential biological markers for PGSVVs. This study aimed to identify aberrant circRNA expression in primary great saphenous vein varicosities (PGSVVs) and investigate its potential clinical significance. High-throughput sequencing was applied to analyze the differential expression of circRNAs in three paired vein samples of PGSVVs and a control group. Computational bioinformatics analyses, including gene ontology (GO) analysis and encyclopedia of genes and genomes (KEGG) pathway analysis, were performed to further enrich microRNA data. Real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to verify the selected aberrant circRNAs in all vein samples, and a receiver operating characteristic (ROC) curve was plotted to evaluate the diagnostic and potential clinical significance. A total of 232 circRNAs were significantly aberrantly expressed in PGSVVs, including 105 and 127 upregulated and downregulated circRNAs, respectively. The predicted top 10 downstream miRNAs of the significantly altered circRNAs are hsa-miR-103a-2-5p, hsa-miR-141-5p, hsa-miR-3692-5p, hsa-miR-4659a-3p, hsa-miR-4659b-3p, hsa-miR-4691-5p, hsa-miR-4778-3p, hsa-miR-6738-3p, hsa-miR-6792-3p and hsa-miR-6873-3p. GO analysis revealed that the most enriched and meaningful biological process, cellular component, and molecular function terms were related to the regulation of catabolic processes, the cytoplasm and ATP binding, respectively. The most enriched and meaningful pathways revealed by KEGG pathway analysis were related to “axon guidance” “vitamin digestion and absorption” and “NF-Kappa B signaling pathway”. Hsa{sub c}irc{sub 0}006427 (P = 0.007), hsa{sub c}irc{sub 0}089810 (P = 0.044) and hsa{sub c}irc{sub 0}005267 (P = 0.049) were significantly downregulated in the PGSVV group relative to their expression levels in the control group, and the area under the ROC curve was 0.9091, 0.8025 and 0.8148, respectively. This study demonstrated that circRNAs were differentially expressed in PGSVVs, and highlights the crucial roles of circRNAs in the pathogenesis of PGSVVs and indicates that hsa{sub c}irc{sub 0}006427, hsa{sub c}irc{sub 0}089810 and hsa{sub c}irc{sub 0}005267 might be potential diagnostic and clinical biological markers.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Alterations of circular RNAs in hyperglycemic human endothelial cells

Circular RNA (circRNA), a family of RNA generated by RNA circularization, is ubiquitously expressed in tissues and possesses increasingly important biological functions. Hyperglycemia-induced endothelial dysfunction is an initiating event in the pathogenesis of diabetes-associated cardiovascular complications. How high glucose may affect circRNAs is unknown. To address this issue, human endothelial cells were exposed to high glucose treatment and the changes of circRNAs were measured by RNA sequencing. A total 3686 circRNAs, including 1040 previously unrecorded circRNAs, were detected; and 95 different expression (DE) circRNAs were observed. The host genes of these DE circRNAs were further studied by function enrichment analyses. These analyses revealed genes of phosphoproteins, transferases, and zine finger proteins. Since circRNAs can function as a microRNA (miRNA) sponge, circRNAs-miRNAs interaction networks were explored by bioinformatics. These analyses identified a number of miRNAs, which might interact with DE circRNAs and play roles in the actions of high glucose on endothelial cells. These results demonstrate that high glucose exposure profoundly changes circRNA expression in endothelial cells. Altered circRNA expression may contribute to the effects of high glucose on endothelial function in diabetes.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Loss of nucleolar localization of NAT10 promotes cell migration and invasion in hepatocellular carcinoma

Highlights: • Residues 68–75 and 989–1018 are complete NuLSs of NAT10. • NuLSs-deleted NAT10 translocates from nucleolus to cytoplasm and membrane. • NuLSs-deleted NAT10 facilitates α-tubulin acetylation, stabilizes microtubule and promotes HCC cell migration. • Cytoplasmic and membranous NAT10 is correlated with poor survival of HCC patients. NAT10, a nucleolar acetyltransferase, participates in a variety of cellular processes including ribosome biogenesis and DNA damage response. Immunohistochemistry staining showed that cytoplasmic and membranous NAT10 is related to the clinical pathologic characteristics in human cancer tissues. However, the mechanism about how NAT10 translocates from the nucleolus to cytoplasm and membrane is unclear. Here, we obtain a NAT10 deletion mutant localizing in cytoplasm and membrane. Bioinformatics analysis showed that residues 68–75 and 989–1018 are two potential nuclear localization signals (NLS) of NAT10. GFP-NAT10 deletion mutant (Δ989-1018) predominantly translocates into cytoplasm with faint signal retained in the nucleolus, while GFP-NAT10(Δ68-75) still remains in the nucleolus and nucleoplasm, indicating residues 989–1018 is the main nucleolar localization signal (NuLS). GFP-NAT10-D3, with both fragments (residues 68–75 and 989–1018) deleted, completely excludes from the nucleolus and translocates to cytoplasm and membrane. Therefore, complete NuLSs of NAT10 should include residues 68–75 and 989–1018. The cytoplasmic and membranous NAT10 mutant (Flag-NAT10-D3) colocalizes with α-tubulin in cytoplasm and with integrin on cell membrane. Importantly, Flag-NAT10-D3 promotes α-tubulin acetylation and stabilizes microtubules. Consequently, Flag-NAT10-D3 promotes migration and invasion in hepatocellular carcinoma (HCC) cells. Statistical analysis of immunohistochemistry staining of NAT10 in HCC tissues demonstrates that the cytoplasmic NAT10 is correlated with poorer prognosis compared with nuclear NAT10, while the membranous NAT10 predicts the poorest clinical outcome of the patients. We thus provide the evidence for the function of cytoplasmic and membranous NAT10 in the metastasis and prognosis of HCC patients.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

A novel small RNA S042 increases acid tolerance in Lactococcus lactis F44

Lactococcus lactis, a gram-positive bacterium, encounters various environmental stresses, especially acid stress, during fermentation. Small RNAs (sRNAs) that serve as regulators at post-transcriptional level play important roles in acid stress response. Here, a novel sRNA S042 was identified by RNA-Seq, RT-PCR and Northern blot. The transcription level of s042 was upregulated 2.29-fold under acid stress by Quantitative RT-PCR (qRT-PCR) analysis. Acid tolerance assay showed that overexpressing s042 increased the survival rate of L. lactis F44 and deleting s042 significantly inhibited the viability under acidic conditions. Moreover, the targets were predicted by online software and four genes were chosen as candidates. Among them, argR (arginine regulator) and accD (acetyl-CoA carboxylase carboxyl transferase subunit beta) were validated to be the direct targets activated by S042 through reporter fusion assay. The regulatory mechanism between S042 and its targets was further investigated through Bioinformatics and qRT-PCR. This study served to highlight the role of the novel sRNA S042 in acid resistance of L. lactis and provided new insights into the response mechanism of acid stress.

60 APPLIED LIFE SCIENCES↗