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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 703 records · Page 39
All sourcesNASA NTRSDOE OSTI
DOE OSTI2018

SIS3, a specific inhibitor of smad3, attenuates bleomycin-induced pulmonary fibrosis in mice

Highlights: • SIS3 ameliorate bleomycin-induced pulmonary fibrosis. • SIS3 reduced phosphorylation of Smad3 in lung of mice treated with bleomycin. • SIS3 suppressed acute lung inflammation induced by bleomycin. Pulmonary fibrosis (PF) is a fatal respiratory disease with no effective medical treatments available. TGF-β/Smads signaling has been implicated to play an essential in the pathogenesis of PF, in which Smad3 act as the integrator of pro-fibrosis signals. In this study, we determined the effect of SIS3, a specific inhibitor of Smad3, in an experimental mouse model of lung fibrosis. We observed that SIS3 treatment significantly reduced bleomycin (BLM)-induced pathological changes and collagen deposition in the lung as indicated by Masson staining, real-time PCR and hydroxyproline content assay. As expected, the levels of Smad3 phosphorylation were decreased in the lung of mice treated with SIS3. Furthermore, SIS3 treatment also suppressed BLM-induced infiltration of inflammatory cells in the lung. Taken together, our results suggest that SIS3 ameliorated BLM-induced PF in mouse lungs. Thus, targeting Smad3 with SIS3 may be an effective approach for treatment of fibrotic disorders.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

SPATULA regulates floral transition and photomorphogenesis in a PHYTOCHROME B-dependent manner in Arabidopsis

Highlights: • SPT promotes floral transition in Arabidopsis. • SPT promotes flowering via FLC, SVP, FT and SOC1. • SPT regulates photomorphogenesis in a PHYB-dependent manner. Light is the most important exogenous stimulus regulating plant growth and various developmental processes. Phytochromes, especially PHYTOCHROME B (PHYB) mediates the various light-mediated processes in Arabidopsis. SPATULA (SPT) is an important transcription factor, which has been reported previously to participate in temperature-mediated transition from seed dormancy to germination. Here we investigate the function of SPT in the floral transition under long day conditions and photomorphogenesis in Arabidopsis. In this study, spt-2 shows significantly delayed flowering time. But mutation of SPT in the background of phyb-1 rescues the phenotype of spt-2. The flowering time of double mutant of spt-2/phyb-1 is similar with the wild type. These results indicate that SPT promotes the transition from vegetative stage to floral stage and it regulates this transition in a PHYB-dependent manner. With qRT-PCR analysis, it is found that SPT regulates flowering time via FLC, SVP, FT and SOC1. Furthermore, SPT also controls photomorphogenesis. spt-2 displays shortened hypocotyls and increased chlorophylls contents compared with the wild type. These phenotypes are also rescued in the double mutant of spt-2/phyb-1. These results indicate that SPT is also involved in photomorphogenic development in Arabidopsis and SPT regulates photomorphogenesis in a PHYB-dependent manner. Collectively, SPT is not only a temperature responder but it is also an important light regulator during plant growth and development.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

l-asparaginase induces intrinsic mitochondrial-mediated apoptosis in human gastric adenocarcinoma cells and impedes tumor progression

Highlights: • We first confirmed that l-asparagine is essential for the growth and proliferation of AGS cells. • We further clarified that l-asparagine deprivation by l-asparaginase promotes anti-proliferation and apoptosis of AGS cells. • We demonstrated that action of l-asparaginase alters mitochondrial membrane permeability and activates intrinsic apoptosis. • We also showed that l-asparaginase impedes tumor progression. l-asparagine essentially regulates growth and proliferation of cancer cells. l-asparaginase is an anti-cancer enzyme that deprives the cancer cells of l-asparagine. The purpose of this study was to explore the mechanism of a novel l-asparaginase from Pseudomonas fluorescens on l-asparagine deprivation mediated anti-proliferation, apoptosis in human gastric adenocarcinoma cells and to evaluate inhibition of angiogenesis. We observed that, the presence of extracellular l-asparagine was essential for the growth of AGS cells. l-asparagine deprivation by l-asparaginase induced metabolic stress, cytotoxicity and apoptosis by G0 phase cell-cycle arrest, modulated the mitochondrial membrane integrity, accelerated caspase-3 activation and instigated DNA damage. The RT-PCR analysis of pro-apoptosis genes: bak1, bax, bbc3, bik, pmaip1, bnip3l, apaf1, casp3, casp7 and casp9 were significantly higher (P xiap, bid, mcl1, and death receptor genes tnf and tradd were significantly down-regulated (P l-asparaginase alters mitochondrial membrane permeability and auxiliary activates intrinsic apoptosis. Therefore, this mechanistic approach might be considered as a targeted enzymotherapy against gastric adenocarcinoma.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Upregulation of lncRNA VDR/CASC15 induced by facilitates cardiac hypertrophy through modulating miR-432-5p/TLR4 axis

Sustained cardiac hypertrophy has threatened human health. With the development of human genome project, non-coding RNAs (ncRNAs) have attracted more and more attentions of researchers. As a subgroup of ncRNAs, long non-coding RNAs (lncRNAs) has been widely studied in human diseases, including cardiac hypertrophy. According to search results of bioinformatics website, lncRNA CASC15 potentially participates in the course of cardiac hypertrophy. According to the result of qRT-PCR, CASC15 expression was upregulated when cardiomyocytes were treated with Ang-II. Moreover, CASC15 was highly expressed in cardiac hypertrophic model. Upregulation of CASC15 was accompanied with some hypertrophic responses. To explore the specific biological function of CASC15 in cardiac hypertrophy, loss-of-function experiments were conducted in Ang-II-induced cardiomyocytes. Results of immunofluorence staining revealed that cell surface area enlarged by Ang-II was decreased when CASC15 was silenced. The expression levels of hypertrophic factors were attenuated by knockdown of CASC15. To detect the molecular mechanism by which CASC15 regulates the progression of cardiac hypertrophy, mechanism experiments were designed and carried out. It was found that CASC15 was activated by the transcription factor VDR. Furthermore, CASC15 can upregulate TLR4 by competitively binding miR-432-5p. In conclusion, Upregulation of lncRNA CASC15 induced by VDR facilitates cardiac hypertrophy via miR-432-5p/TLR4 axis.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

miR-410–3p promotes prostate cancer progression via regulating PTEN/AKT/mTOR signaling pathway

Prostate cancer has become one of commonest urologic tumors in male. In recent years, miRNAs are continually attracting attentions of researchers for their special regulatory function in human cancers. Previous study has revealed that miR-410 acts as a biomarker for the diagnosis of prostate cancer. Whereas, the specific biological function of miR-410–3p in prostate cancer remains unknown. The aim of this study is to explore the function and molecular mechanism of miR-410–3p in prostate cancer. The high expression of miR-410–3p was examined in prostate cancer tissues and cell lines by qRT-PCR. Next, the prognostic value was identified by Kaplan Meier method. High expression of miR-410–3p indicated poor prognosis of prostate cancer patients. To investigate the biological function of miR-410–3p in prostate cancer, loss-of function assays were designed and performed in two prostate cancer cell lines (PC3 and DU145). As a result, downregulated miR-410–3p suppressed cell proliferation, migration and EMT progress. Moreover, flow cytometry analysis was performed to determine that the acceleration effects of miR-410–3p on cell apoptosis. Mechanistically, further analysis demonstrated that the effects of miR-410–3p exert oncogenic functions through downregulating PTEN. All findings in this study revealed that miR-410–3p inhibits prostate cancer progression via downregulating PTEN/AKT/mTOR signaling pathway.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

ZKSCAN3 promotes breast cancer cell proliferation, migration and invasion

Highlights: • ZKSCAN3 is upregulated in breast cancer tissue and cell lines. • Knocking down ZKSCAN3 suppresses breast cancer cell proliferation, enhances cell apoptosis and blocks cell cycle. • Knocking down ZKSCAN3 significantly inhibits breast cancer cell migration and invasion. • Knocking down ZKSCAN3 significantly inhibits breast tumor growth in vivo. • Knocking down ZKSCAN3 reduces activity of the Akt/mTOR pathway. ZKSCAN3, a zinc-finger transcription factor, which has been shown to be upregulated in several human cancer. However, the expression level, function and mechanism of ZKSCAN3 in breast cancer remains unknown. In the current study, immunohistochemistry, western blot and quantitative real time polymerase chain reaction (qRT-PCR) results showed that ZKSCAN3 was overexpressed in breast cancer tissue compared with normal breast tissue. Through analyzing the clinicopathological characteristics, we demonstrated that positive ZKSCAN3 expression predicted poor prognosis of patients with breast cancer. The expression level of ZKSCAN3 protein/mRNA in breast cancer cells (MCF-7 and MDA-MB-231) was higher than its expression in normal breast cells (HBL-100). Knocking down ZKSCAN3 via its short hairpin RNA (shRNA) in MCF-7 and MDA-MB-231 inhibited cell viability, migration and invasion. Western blot analysis showed that ZKSCAN3 silencing lead to significant decreases in the expression of Cyclin D1, B-cell lymphoma-2 (Bcl-2), and matrix metalloproteinase (MMP)-2/MMP-9, as well as increases in the expression of Bcl2 Associated X Protein (Bax) in breast cancer cells. Additionally, ZKSCAN3-shRNA expression markedly suppressed tumor growth in breast cancer xenograft mice. Finally, we demonstrated that silencing of ZKSCAN3 was able to inhibit Akt/mTOR signaling pathway by blocking p-Akt and p-mTOR protein expression in breast cancer cells. These results demonstrate that ZKSCAN3 plays a significant role in the progression of breast cancer. Therefore, ZKSCAN3 is a potential therapeutic target for breast cancer.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Circular RNA circ{sub 0}001649 acts as a prognostic biomarker and inhibits NSCLC progression via sponging miR-331-3p and miR-338-5p

Accumulating evidence documented the key functions of circular RNAs (circRNAs) in various malignancies. However, the study regarding the involvement of circRNAs in non-small cell lung cancer (NSCLC) has just begun. In the present study, qRT-PCR was used to determine the expression of circ{sub 0}001649 in NSCLC tissues and cells. Its clinical significance was further assessed by Fisher's exact test, Kaplan-Meier analysis and Cox regression model. Additionally, loss-of-function and gain-of-function experiments were carried out to detect the functional role of circ{sub 0}001649 in cell proliferation, migration and invasion. Furthermore, animal study was performed to confirm the in vitro results. Importantly, luciferase reporter assay was induced to reveal the underlying mechanism of circ{sub 0}001649. As a result, circ{sub 0}001649 was decreased in NSCLC tissues and cells and this downregulation is correlated with advanced TNM stage, positive lymph node metastasis and unfavorable prognosis. Additionally, circ{sub 0}001649 inhibited cell growth and metastasis both in vitro and in vivo. In mechanism, circ{sub 0}001649 was identified as the sponge of miR-331-3p and miR-338-5p. Moreover, the biological functions of circ{sub 0}001649 is partly dependent on its regulation on miR-331-3p and miR-338-5p. Collectively, this study suggested that circ{sub 0}001649/miR-331-3p/miR-338-5p regulatory signaling might be a potential target for NSCLC therapy.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Autophagy induced by type III secretion system toxins enhances clearance of Pseudomonas aeruginosa from human corneal epithelial cells

Highlights: • Ocular P. aeruginosa isolates induce autophagy in human corneal epithelial cells. • T3SS mutants were defective in inducing autophagy and becn1 expression. • T3SS (−)ve strains were less sensitive to external autophagy modulators. • Pro-autophagic role of T3SS regulates intracellular survival of P. aeruginosa. Corneal ulcers caused by Pseudomonas aeruginosa may lead to severe visual disability due to impaired bacterial clearance from corneal tissues. Our purpose was to study the role of autophagy in the intracellular clearance of P. aeruginosa from human corneal epithelial cells (HCET) and its regulation by the bacterial type III secretion system (T3SS) toxins. Nine different corneal ulcer isolates of P. aeruginosa, PAO1 and T3SS mutants of PAO1 were used to infect HCET cells. Induction of autophagy (Immunofluorescence and Western blot) and pro-inflammatory gene expression (real time PCR) by P. aeruginosa and the role of autophagy in intracellular bacterial clearance were studied in the context of T3SS genotypes. The clinical isolates and PAO1 induced autophagy irrespective of the T3SS genotype, whereas the T3SS mutants were relatively defective in inducing autophagy and becn1 gene expression. External induction of autophagy significantly reduced the intracellular load of P. aeruginosa strains that were associated with worst clinical outcomes. The T3SS negative isolate and PAO1ΔexoST were less sensitive to pharmacological modulation of autophagy and had relatively higher replication potential, suggesting a possible mechanism of bacterial survival in the absence of T3SS toxins. Overall, our results highlight a selective role for autophagy in bacterial clearance from corneal epithelial cells and emphasize the pro-autophagic role of bacterial toxins in the context of corneal ulcers.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Saturated fatty acids-induced miR-424–5p aggravates insulin resistance via targeting insulin receptor in hepatocytes

Highlights: • miR-424–5p expression is upregulated by SFA or HFD-induced obesity in hepatocytes. • Mechanistically, miR-424–5p targets 3′UTR of INSR directly, thereby downregulating INSR at the post-transcriptional step. • The ectopic expression of miR-424–5p impairs insulin signaling and insulin-induced glycogen synthesis in hepatocytes. • This study highlights the potential role of miR-424–5p in the insulin signaling pathway through the repression of INSR. The excessive intake of saturated fatty acids (SFA) causes obesity and liver steatosis, which are major risk factors for insulin resistance and type 2 diabetes. Although the expression of certain microRNAs (miRNAs) targeting the insulin signaling molecules are regulated aberrantly in SFA-induced obesity, their implications on hepatic insulin resistance are largely unknown. This study examined the associations of miR-424–5p, which is induced by SFA, with the development of insulin resistance. SFA palmitate (PA)-treated HepG2 cells and high fat diet (HFD)-induced obese mouse livers showed an impairment of insulin signaling due to a significant decrease in INSR and IRS-1 expression. Based on expression profiling and qRT-PCR analysis, miR-424–5p, which presumably targets the 3′UTR of INSR, was upregulated in both PA-treated HepG2 cells and the liver of HFD-fed mice. miR-424–5p was found to target the 3′UTR of INSR directly and downregulated INSR expression at the post-transcriptional step. Furthermore, the overexpression of miR-424–5p suppressed INSR expression significantly, leading to impaired insulin signaling and glycogen synthesis in hepatocytes. A novel mechanism for how SFA-induced miR-424–5p impairs insulin signaling through the targeting of INSR is reported. In addition, the crucial role and underlying mechanism of miR-424–5p in the obesity-induced hepatic insulin resistance is explained.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Up-regulation of TRAF2 inhibits chondrocytes apoptosis in lumbar facet joint osteoarthritis

Tumor necrosis factor receptor-associated factor 2 (TRAF2) has been demonstrated that it plays a significant role in cell death receptor signal transduction. The purpose of this study was to investigate the expression of TRAF2 and its possible role in FJOA. We observed an up-regulation of TRAF2 in FJOA by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR) compared to normal tissues. In vitro, we used TNF-α to stimulate Human SW1353 chondrosarcoma cells to establish the chondrocytes injury model. Western blot analysis revealed significant expression of TRAF2 and cleaved caspase-3/8 in SW1353 cells. Co-localization of TRAF2/cleaved caspase-3/8 was detected in the cells injury model by double-labeling immunofluorescent staining. We demonstrated a possible anti-apoptotic effect of TRAF2 in chondrocyte apoptosis in FJOA by knockdown of its expression with siRNA. Moreover, TRAF2 knockdown was demonstrated to enhance TNF-α-induced apoptosis by flow cytometry assay. In conclusion, our results show that the up-regulation of TRAF2 may play an important role in the inhibition of chondrocyte apoptosis of FJOA.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Upregulated circular RNA circ{sub 0}016760 indicates unfavorable prognosis in NSCLC and promotes cell progression through miR-1287/GAGE1 axis

Non-small cell lung cancer (NSCLC) is an aggressive malignancy with increasing worldwide incidence and is characterized by dismal prognosis due to its early recurrence and metastasis. Accumulating evidence documented that aberrantly expressed circular RNAs (circRNAs) are critically involved in tumorigenesis. In the current study, we focused on a novel circRNA, circ{sub 0}016760 in NSCLC. qRT-PCR was carried out to determine the expression level of circ{sub 0}016760 in tissue samples and cell lines. The clinical value of circ{sub 0}016760 was also investigated. The functions of circ{sub 0}016760 was explored by CCK-8, colony formation, flow cytometry, Transwell and animal experiments. Luciferase reporter assays were conducted to investigate the association between circ{sub 0}016760 and miR-1287 and miR-1287 and GAGE1. We found that circ{sub 0}016760 was enhanced in NSCLC tissues and cells and the upregulation of circ{sub 0}016760 is associated with advanced TNM stages, lymph node metastasis and adverse prognosis in NSCLC patients. Moreover, circ{sub 0}016760 could significantly promote cell growth and metastatic properties and inhibit cell apoptosis in NSCLC cells. In mechanism, circ{sub 0}016760 functioned as a sponge for miR-1287 and regulated the expression of GAGE1. Subsequently, functional assays illustrated that the oncogenic properties of circ{sub 0}016760 is partly attribute to the regulation of miR-1287/GAGE1 axis. In summary, circ{sub 0}016760/miR-1287/GAGE1 signaling might play vital roles in the tumorigenesis and progression of NSCLC.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Overexpression of circular RNA circ{sub 0}01569 indicates poor prognosis in hepatocellular carcinoma and promotes cell growth and metastasis by sponging miR-411-5p and miR-432-5p

Emerging evidence indicated that abnormally expressed circular RNAs (circRNAs) are critically involved in tumorigenesis and development of several cancers. However, the study relevant to the relationship between circRNAs and hepatocellular carcinoma (HCC) is rare. In this study, the expression of circ{sub 0}01569 in HCC tissue specimens and cells were determined by qRT-PCR. The clinical relevance of circ{sub 0}01569 was also analyzed. In addition, loss-of-function and gain-of-function assays were conducted to explore the biological functions of circ{sub 0}01569. Furthermore, tumor formation and lung metastasis studies were induced to confirm the in vitro data. Mechanistically, bioinformatics analysis and luciferase reporter assays were conducted to illustrate the underlying mechanism of circ{sub 0}01569. The results indicated that circ{sub 0}01569 was overexpressed in HCC tissue samples and cells. This overexpression is correlated with larger tumor size, advanced TNM stages and unfavorable prognosis in the patients with HCC. Additionally, circ{sub 0}01569 significantly facilitated HCC cell growth, migration and invasion. The animal studies further confirmed the in vitro results. More importantly, circ{sub 0}01569 could directly sponge miR-411-5p and miR-432-5p. The oncogenic functions of circ{sub 0}01569 is partially dependent on its regulation on miR-411-5p and miR-432-5p. In summary, circ{sub 0}01569/miR-411-5p/miR-432-5p regulatory signaling might be a rational HCC-related therapeutic target.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

MiR-590-5p alleviates intracerebral hemorrhage-induced brain injury through targeting Peli1 gene expression

Highlights: • Down-regulation of miR-590-5p in ICH mice and LPS-induced microglia cells. • MiR-590-5p attenuates ICH-induced brain injure. • Peli1-1 is a direct target of miR-590-5p. • Peli1 overexpression abrogates the effect of miR-590-5p mimic on ICH-induced brain injure. Intracerebral haemorrhage (ICH) is a common and devastating cerebrovascular disease with high morbidity and mortality, and its pathophysiological mechanisms were complex and still unclear. Increasing researches reported that microRNAs (miRNAs) played an important role in ICH-induced brain injury and microglial activation. In this study, we investigated the biological function of miR-590-5p and explored its molecular mechanism in ICH mice. The results of qRT-PCR showed that miR-590-5p expression level was down-regulated in perihematomal brain samples of ICH mice compared with that of sham group. In LPS-induced microglia cells, miR-590-5p level was also down-regulated at 24 h post-LPS compared with that of control group. Moreover, miR-590-5p overexpression remarkably increased the cerebral water content and neurological severity scores compared with that of scramble group in ICH mice. The production of inflammatory cytokine including IL-6, interleukin (IL)-1β, and tumor necrosis factor (TNF)-α in ICH mice was notably inhibited by miR-590-5p overexpression. Furthermore, the results of dual-luciferase reporter assay indicated that Pellino-1 (Peli1) was a direct target of miR-590-5p. MiR-590-5p overexpression remarkably inhibited the Peli1 gene expression both mRNA and protein levels. In addition, Peli1 overexpression partly abrogated the inhibitory effect of miR-590-5p mimic. Taken together, these datas suggested that miR-590-5p attenuated brain injury in ICH mice through inhibiting Peli1 gene expression, indicating that miR-590-5p may be a promising molecular target for ICH treatment.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

CCN3 secreted by prostaglandin E2 inhibits intimal cushion formation in the rat ductus arteriosus

Highlights: • CCN3 is secreted from smooth muscle cells of the ductus arteriosus by prostaglandin E{sub 2}. • Nano-flow LC-MS/MS analysis is a powerful tool for detecting a novel secreted protein. • CCN3 inhibits intimal cushion formation in the ductus arteriosus. The ductus arteriosus (DA), an essential fetal shunt between the pulmonary trunk and the descending aorta, changes its structure during development. Our previous studies have demonstrated that prostaglandin E{sub 2} (PGE{sub 2})-EP4 signaling promotes intimal cushion formation (ICF) by activating the migration of DA smooth muscle cells via the secretion of hyaluronan. We hypothesized that, in addition to hyaluronan, PGE{sub 2} may secrete other proteins that also regulate vascular remodeling in the DA. In order to detect PGE{sub 2} stimulation-secreted proteins, we found that CCN3 protein was increased in the culture supernatant in the presence of PGE{sub 2} in a dose-dependent manner by nano-flow liquid chromatography coupled with tandem mass spectrometry analysis and enzyme-linked immunosorbent assay. Quantitative RT-PCR analysis revealed that PGE{sub 2} stimulation tended to increase the expression levels of CCN3 mRNA in DA smooth muscle cells. Immunohistochemical analysis revealed that CCN3 was highly localized in the entire smooth muscle layers and the endothelium of the DA. Furthermore, exogenous CCN3 inhibited PGE{sub 2}-induced ICF in the ex vivo DA tissues. These results suggest that CCN3 is a secreted protein of the DA smooth muscle cells induced by PGE{sub 2} to suppress ICF of the DA. The present study indicates that CCN3 could be a novel negative regulator of ICF in the DA to fine-tune the PGE{sub 2}-mediated DA remodeling.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Wound-induced polypeptides improve resistance against Pseudomonas syringae pv. tomato DC3000 in Arabidopsis

Wound-induced polypeptides (WIPs) are a novel class of polypeptides with the length less than 100 amino acids. Our previous research has identified a number of WIP genes in soybean (Glycine max) root nodules. However, functions of WIPs in planta remains largely unknown. Here, we identified five WIP-encoding genes, AtWIP1-5, in Arabidopsis. Among them, AtWIP1 and -2 are ubiquitously expressed in a partially overlapping pattern as revealed by both qRT-PCR and promoter:GUS assays. Subcellular localization analyses reveal that both AtWIP1 and -2 are localized at the plasma membrane while AtWIP1 shows a punctate distribution pattern. AtWIP1, -2 are transcriptionally induced by flg22 treatment, but repressed by effector(s) of Pseudomonas syringae pv. tomato DC3000 (Pst DC3000). Heterologous overexpression of GmWIP genes enhances resistance of Arabidopsis to Pst DC3000 at the cost of growth inhibition. Moreover, overexpression of GmWIP genes promotes pattern-triggered immunity (PTI) evidenced by increased expressions of flg22-inducible genes and enhanced seedling growth inhibition under flg22 treatment. Taken together, our results indicate that WIPs positively regulate plant resistance against Pst DC3000 by enhancing PTI responses.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Pigment epithelium derived factor (PEDF) expression in the male tract of Wistar rats

Highlights: • PEDF expression is detected in epididymal tract of male rats. • PEDF expression was not detected in rat testes. • PEDF expression was decreased by using flutamide. • PEDF expression was recovered after flutamide cessation. Pigment epithelium derived factor (PEDF) expression has been described in many organs as showing neurotrophic, anti-angiogenic, anti-apoptotic, anti-inflammatory, anti-oxidant and pro-cell survival properties. However, references to its activity in the male reproductive system are scarce. We aimed to characterize the expression of PEDF in the male reproductive tract of Wistar rats by using RT-PCR, western blot and immunostaining and also evaluate the effect of flutamide in PEDF expression. We found that PEDF is expressed in the epididymis, prostate and seminal vesicles in Wistar rats, but notably not in the testes. Under the effect of flutamide PEDF expression decreased, recovering by suppressing the antiandrogen. The epididymis is an essential organ in sperm maturation-storages. The role of PEDF in this physiological process has not been fully elucidated yet, but considering that in other systems PEDF has anti-apoptotic, anti-oxidants and pro-cell survival properties, its expression along the epididymis could play a role in the protection of spermatozoa while they are stored.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Grhl2 regulation of SPINT1 expression controls salivary gland development

Development of the salivary gland is characterized by extensive branching morphogenesis and lumen formation, the latter of which is closely associated with differentiation into acinar and ductal cells. Although various molecules, including signaling and cell adhesion molecules, have been implicated in salivary gland development, transcription factors (TFs) regulating the expression of those molecules and morphological development of the gland are largely unknown. Here we show that knockdown of the epithelial TF, Grainyhead-like 2 (Grhl2), with siRNA in developing mouse submandibular salivary gland (SMG) cultured ex vivo resulted in retardation of epithelial development. This retardation was concomitant with suppression of gene expression for the cell adhesion molecules, such as E-cadherin and the extracellular protease inhibitor SPINT1, and with the disorganized deposition of the basal lamina protein laminin. ChIP-PCR demonstrated the binding of Grhl2 protein to the Spint1 gene in the SMG. Notably, addition of recombinant SPINT1 protein in cultured SMG overcame the suppressive effects of Grhl2 siRNA on epithelial development and laminin deposition. These findings show that Grhl2 regulation of SPINT1 expression controls salivary gland development.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

As an independent prognostic factor, USP6 promotes the invasion and metastasis of colon cancer

Highlights: • USP6 is highly expressed in colon cancer tissues at both mRNA and protein levels. • High USP6 expression was closely associated with the invasion and metastasis of colon cancer. • USP6 overexpression promotes colon cancer cell invasion and metastasis both in vitro and in vivo. • USP6 is considered as a novel gene in promoting the invasion and metastasis of colon cancer. The lack of efficient tumor invasion and metastatic biomarkers led to high mortality rates in colon cancer patients. Aberrant expression of ubiquitin-specific protease 6 (USP6) was involved in several diseases including cancer, while its role in the progression of colon cancer was still unclear. In this study, USP6 was evaluated at both mRNA and protein levels by using RT-PCR, western blot and immunohistochemistry staining analyses. The results revealed that high USP6 expression predicted poor disease-specific survival and overall survival through Kaplan-Meier analyses with log-rank tests, univariate and multivariate Cox analyses. Furthermore, cell function assay demonstrated that USP6 could promote colon cancer cells’ invasion in vitro and liver metastasis in vivo. These findings indicated that high USP6 expression contributed to the progression of colon cancer and USP6 may be a valuable prognostic factor in patients with colon cancer.

60 APPLIED LIFE SCIENCES↗