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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 631 records · Page 35
All sourcesNASA NTRSDOE OSTI
DOE OSTI2017

Circular RNA hsa-circ-0010729 regulates vascular endothelial cell proliferation and apoptosis by targeting the miR-186/HIF-1α axis

Circular RNAs (circRNAs) are a group of non-protein-coding RNAs generated from back splicing. Emerging evidence has demonstrated its vital regulation on angiogenesis. However, the underlying mechanism responsible for circRNAs effects on vascular endothelial cells is still unclear. In the present study, we screened the expression profiles and investigated the physiological role of circRNAs in hypoxia-induced human umbilical vein endothelial cells (HUVECs). Using circRNA microarray analysis, we identified 36 circRNAs that were significantly dysregulated including 14 down-regulated circRNAs and 22 up-regulated with 2-fold change (P < 0.05). From the over-expressed circRNAs, hsa-circ-0010729 was selected as candidate circRNA and which was validated to be significantly up-regulated using RT-PCR. In loss-of-function experiments of HUVECs, hsa-circ-0010729 knockdown suppressed the proliferation and migration ability and enhanced apoptosis. Bioinformatic prediction and luciferase assay revealed that hsa-circ-0010729 and hypoxia inducible factor 1 alpha (HIF-1α) were targeted by miR-186. Validation experiments verified that hsa-circ-0010729 was co-expressed with HIF-1α, being negatively correlated with miR-186. Moreover, rescue experiments demonstrated that miR-186 inhibitor could reverse the role of hsa-circ-0010729 knockdown on HUVECs progression. Overall, the present study identifies the crucial regulation of hsa-circ-0010729 on vascular endothelial cell proliferation and apoptosis via targeting miR-186/HIF-1α axis. - Highlights: • Circular RNA expression profile identifies hsa-circ-0010729 is up-regulated in hypoxia-induced HUVECs. • Hsa-circ-0010729 knockdown suppresses the proliferation and migration ability, and enhanced the apoptosis. • Hsa-circ-0010729 acts as an endogenous sponge of miR-186. • HIF-1α is targeted by miR-186. • Hsa-circ-0010729 regulates vascular endothelial cells proliferation and apoptosis via targeting miR-186/HIF-1α axis.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

MiR-129-5p inhibits non-small cell lung cancer cell stemness and chemoresistance through targeting DLK1

Non-small cell lung cancer (NSCLC) stemness and chemoresistance represent significant barriers to successful treatment. Downregulation of microRNA 129 (miR-129) has been implicated in a variety of cancers, and miR-129-5p has recently been shown to reduce lung cancer proliferation and metastasis. In this study, we used NSCLC cell lines A549 and H460 to investigate the effects of miR-129-5p on NSCLC stemness and chemoresistance. CD133 + stem cells in both lines showed reduced miR-129-5p expression, and introducing miR-129-5p into these cells reduced stem cell markers and self-renewal ability, as measured by qRT-PCR, western blot, MTT assays, and sphere formation assays. MTT, colony formation, and DAPI staining assays further revealed that miR-129-5p inhibits NSCLC chemoresistance when the cells are treated with varying doses of CDDP. The target prediction algorithm TargetScan allowed us to identify notch signaling receptor delta-like 1 homolog (DLK1) as a potential target for miR-129-5p in its inhibition of NSCLC stemness and chemoresistance. Indeed, our DLK1 luciferase reporter plasmid demonstrated that miR-129-5p reduces DLK1 expression. Furthermore, DLK1 overexpression partially rescued inhibition of NSCLC stemness and chemoresistance caused by miR-129-5p. Overall, these results demonstrate that miR-129-5p inhibits NSCLC stemness and chemoresistance through direct targeting of DLK1. They suggest that miR-129-5p and DLK1 can play a role in an effective treatment strategy for NSCLC.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

Macrophage infiltration regulates the adipose ECM reconstruction and the fibrosis process after fat grafting

Background: Fat grafting experiences a regeneration process from free lipoaspirate to intact adipose tissue. The adipose extracellular matrixes (ECM) provide the structure and biochemical support for surrounding cells; inflammatory cells, like macrophages, regulate the process. Our hypothesis states that transferred fat undergoes ECM remodeling after fat grafting and this process is regulated by macrophage infiltration. Methods: Lipoaspirate was injected subcutaneously into the back of nude mice. The micro-structure of the fat grafts was observed and evaluated using scanning electron microscope (SEM) and collagen I immunohistostaining. The gene transcription level of collagen proteins and the matrix metalloproteinases (MMPs) were assessed by qRT-PCR. Local injection of clodronate-encapsulated liposome was used to evaluate the role of macrophages of fat grafts at different stages in ECM remodeling, depletion of macrophages, at different time points (Week 1 and Week 4). Results: Results from the SEM analysis showed that liposuction caused severe damage to the ECM structure in freshly aspirated adipose tissue. On Day 1 post-transplantation, the surface of adipocytes was covered with platelets and this secreted fibrin network on the fat grafts. An integral adipose structure was already established with an intact ECM at the end of Week 1. The early depletion of macrophages remarkably hindered ECM reconstruction process by down-regulating the expression of collagen proteins and MMPs. Expression of Collagen I was significantly decreased after depletion of macrophages in both gene and protein levels. Results also showed that the depletion of macrophages at the later stage of fat grafting resulted in less fibrosis and capsule formation. Conclusion: Free fat aspirates undergo a prompt ECM reconstruction process and completed in the first week; this process can be initiated with platelets and mainly modulated by inflammatory cells such as macrophages. It was also observed that prolonged macrophage infiltration contributes to fibrosis and capsule formation in fat grafts. - Highlights: • Free fat aspirates undergo a prompt ECM reconstruction process in the first week. • The ECM reconstruction process is initiated with platelets and modulated by macrophages. • Depletion of macrophage in transferred fat resulted in delayed ECM reconstruction.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

Upregulated long non-coding RNA LOC90784 promotes cell proliferation and invasion and is associated with poor clinical features in HCC

A growing amount of literature has indicated that long non-coding RNAs (lncRNAs) are important factors in hepatocellular carcinoma (HCC) progression. However, the significance of lncRNAs in the progression and prognosis of liver cancer is largely unknown. In the present study, upregulated lncRNA LOC90784 was identified through integrative analysis of (GSE58043) and (GSE55191). Furthermore, associations between LOC90784 expression and the clinicopathological characteristics of patients were analyzed with a validated cohort 1 and the Cancer Genome Atlas (TCGA) cohort 2. We investigated the mechanisms by which this highly expressed lncRNA promotes HCC proliferation, invasion and migration via qRT-PCR, fluorescence in situ hybridization (FISH) staining, siRNA transfection, cell proliferation assays, Transwell and colony formation assays, flow cytometry analysis and Western blot. The results showed that LOC90784 expression levels were significantly higher in HCC cell lines and tissues and mainly localized in the cytoplasm. Knockdown of lncRNA LOC90784 expression inhibited proliferation and induced apoptosis and cell cycle arrest by promoting Bax and repressing CDK4 and Cyclin D1 protein expression; it also inhibited invasion and migration by repressing MMP2 and MMP9 expression in HCC cells. LOC90784 overexpression was associated with poor clinical features in the two cohorts and poor overall survival rates in HCC patients with clear resection margins (R0) in cohort 2. These results indicated that LOC90784 upregulation may be a critical oncogene and potential new biomarker in HCC. - Highlights: • LncRNA LOC90784 was overexpressed in HCC tissues and cell lines. • Highly expressed lncRNA LOC90784 was associated with poor clinical features in HCC. • Knockdown of lncRNA LOC90784 expression inhibited proliferation and induced apoptosis and cell cycle arrest. • Knockdown of lncRNA LOC90784 expression inhibited invasion and migration.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

miR-137 attenuates Aβ-induced neurotoxicity through inactivation of NF-κB pathway by targeting TNFAIP1 in Neuro2a cells

Background: Accumulation of β-amyloid (Aβ) and neuroinflammation are implicated in the pathogenesis and development of Alzheimer's disease (AD). Neuron-enriched miR-137 was aberrantly downregulated and may be associated with the pathogenesis of AD. However, the detailed function of miR-137 in AD pathogenesis and the molecular mechanism have not been elucidated. Methods: The expressions of miR-137 and tumor necrosis factor alpha (TNFα)-induced protein 1 (TNFAIP1) at mRNA and protein levels in primary mouse cortical neurons and Neuro2a (N2a) cells exposed to different concentrations of Aβ{sub 25-35} were examined by qRT-PCR and western blot. Luciferase reporter assay was used to confirm the potential target of miR-137. MTT assay, flow cytometry analysis, caspase-3 activity assay, Enzyme-linked immunosorbent assay (ELISA), and western blot were used to detect cell viability, apoptosis, caspase-3 activity, Nuclear factor-kappa B (NF-κB) activity and level, respectively. Results: Aβ{sub 25-35} downregulated miR-137 and upregulated TNFAIP1 in primary mouse cortical neurons and N2a cells. In addition, miR-137 was found to directly target TNFAIP1 and suppress its mRNA and protein levels. Moreover, miR-137 restoration and TNFAIP1 knockdown facilitate Aβ{sub 25-35}-induced cell toxicity, apoptosis, caspase-3 activity, and activated NF-κB in N2a cells, which was partially abolished by TNFAIP1 overexpression. Conclusion: miR-137 attenuated Aβ-induced neurotoxicity through inactivation of NF-κB pathway by targeting TNFAIP1 in N2a cells, shedding light on the molecular mechanism of miR-137 underlying Aβ-induced neurotoxicity. - Highlights: • Aβ{sub 25-35} downregulated miR-137 and upregulated TNFAIP1 in cortical neurons and N2a cells. • miR-137 suppressed TNFAIP1 expression through direct binding at the 3′UTR. • miR-137 overexpression contributed to Aβ{sub 25-35}-induced cell toxicity and apoptosis. • miR-137 overexpression on Aβ{sub 25-35}-induced neurotoxicity activated NF-κB pathway.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

Celecoxib induced apoptosis against different breast cancer cell lines by down-regulated NF-κB pathway

Inducible cyclooxgenase-2 (COX-2) is commonly overexpressed in breast tumors and is a target for cancer therapy. Here, we studied the effect of Celecoxib, a COX-2 inhibitor on two molecular breast cancer subtypes—MDA-MB-231 and SK-BR-3. Firstly, MDA-MB-231 and SK-BR-3 cells were treated with various concentration of Celecoxib for 24 and 48 h. Celecoxib-inhibition of NF-κB (p52 and p65) transcriptional activity and effect of Caspase 3 pathway were examined by western blotting. COX-2 mRNA was assessed by RT-PCR. Cell viability was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazoliumbromide (MTT) assay. Both cell cycle profiles and apoptosis were analyzed using flow cytometry. We found that Celecoxib inhibited the proliferation of the MDA-MB-231 cell line in a dose-time dependent manner versus SK-BR-3 in a dose dependent manner only (p < 0.05). Celecoxib induced apoptosis of the MDA-MB-231 and SK-BR-3 cell lines in a dose-time dependent manner (p < 0.05) with more mean apoptotic cells in MDA-MB-231 than SK-BR-3. Significant cell-cycle arrest at the G1 phase in the MDA-MB-231 versus G2 phase in SK-BR-3 cell lines. NF-κB (p52 and p65) and COX-2 expressions were downregulated in a dose dependent manner, while Caspase 3 expression was upregulated in both cell lines. In this present study, our data indicated Celecoxib might affect each breast cancer subtype independently. Therefore, when using Celecoxib in treatment of breast cancer, it is imperative to consider the subtype of breast cancer on a molecular level. - Highlights: • We assumed and proved the correlation of levels of caspase-3, NF-κB (p52, p65) expression in the two types of breast cancer cells (i.e. MDA-MB-231 and SKBR3), and to determine whether exposure to different concentrations of Celecoxib has an impact in expression of these apoptotic and anti-apoptotic pathways respectively. • The difference in effect of Celecoxib was investigated in the two molecular subtypes of breast cancer cells i.e. MDA-MB-231 (ER–, PR–, HER2–) and SKBR3 (ER–, PR–, HER2+ Ki67 high) with regard to proliferation and cell cycle analysis. • In our study, we demonstrated that when using Celecoxib in treatment of breast cancer, it is imperative to consider the subtype of breast cancer on a molecular level.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

MicroRNA-193-5p modulates angiogenesis through IGF2 in type 2 diabetic cardiomyopathy

Background: Patients with diabetic cardiomyopathy are often associated with increasing risk of heart failure. In this work, we used animal model to characterize the angiogenic effect of microRNA-193-5p, miR-193-5p in type 2 diabetic Goto-Kakizaki (GK) rats' myocardial microvascular endothelial cells, MMEC(GK). Methods: MiR-193-5p in MMEC(GK) was compared to its expression in Wistar rat MMEC. In MMEC(GK), miR-193-5p was downregulated through viral infection. Its angiogenic effects on MMEC(GK) migration and proliferation were assessed by transwell and MTT assays, respectively. Downstream target of miR-193-5p, insulin growth factor 2 (IGF2), was assessed by dual-luciferase activity, qRT-PCR and western blot assays, respectively. In miR-193-5p-downregulated MMEC(GK), IGF2 was further de-regulated to assess its mechanism in miR-193-5p-downreuglation induced angiogenic regulation. Results: MiR-193-5p is overexpressed in MMEC(GK). Its downregulation has significantly angiogenic effect by inducing migration and proliferation in MMEC(GK). IGF2 was demonstrated to be directly regulated by miR-193-5p in MMEC(GK). In addition, IGF2 inhibition in miR-193-5p-downregulated MMEC(GK)’s severely hindered cell migration and proliferation. Conclusion: MiR-193-5p is an active angiogenic factor in diabetic cardiomyopathy, possibly through inverse regulation on its downstream IGF2 gene.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

LncRNA-SNHG1 contributes to gastric cancer cell proliferation by regulating DNMT1

Objective: The objective of this study was to determine the expression of long-chain non-coding RNA SNHG1 (lncRNA-SNHG1) in gastric carcinoma and explore its function on cancer cell proliferation. Methods: The expression of lncRNA-SNHG1 in tumor tissues and corresponding adjacent tissues from 50 patients with gastric cancer was detected with realtime-PCR. The relationships between the expression of lncRNA-SNHG1 and clinicopathological features of gastric cancer patients were analyzed. Survival analysis was performed to study the correlation between lncRNA SNHG1 expression and patient prognosis. To assess the effect of LncRNA SNHG1 on proliferation in cancer cells, cell viability and colony formation assays were conducted when lncRNA SNHG1 was upregulated or downregulated by Lentivirus or plasmid in gastric cancer cells. Furthermore, in vivo tumor assay was performed to confirm the impact of lncRNA SNHG1 on proliferation of gastric cancer. Results: The expression of lncRNA SNHG1 in gastric cancer tissues was significantly higher than that in adjacent tissues and was correlated with TNM stage, T stage, and lymph node metastasis. The survival time of patients with higher expression level of lncRNA-SNHG1 was significantly lower than that of the lower expression level. LncRNA-SNHG1 accelerated the proliferation of gastric cancer cells obviously and increased the expression of DNMT1. Conclusion: LncRNA SNHG1 promotes DNMT1 expression, which facilitates the gastric cancer proliferation. - Highlights: • The expression of lncRNA SNHG1 in gastric cancer tissues was significantly higher than that in adjacent tissues. • The expression of lncRNA SNHG1 was correlated with TNM stage, T stage, and lymph node metastasis. • LncRNA-SNHG1 accelerated the proliferation of gastric cancer cells, both in vitro and in vivo. • LncRNA-SNHG1 promoted DNMT1 expression, which facilitated the gastric cancer proliferation.

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

Curcumin inhibits bladder cancer stem cells by suppressing Sonic Hedgehog pathway

Cancer stem cells (CSCs) is responsible for the recurrence of human cancers. Thus, targeting CSCs is considered to be a valid way for human cancer treatment. Curcumin is a major component of phytochemicals that exerts potent anticancer activities. However, the effect of curcumin on bladder cancer stem cells (BCSCs) remains to be elucidated. In this study, we investigated the mechanism of curcumin suppressing bladder cancer stem cells. In this study, UM-UC-3 and EJ cells were cultured in serum-free medium (SFM) to form cell spheres that was characterized as BCSCs. Then cell spheres were separately treated with different concentrations of curcumin and purmorphamine. Cell cycle analysis were used to determine the percentage of cells in different phases. Western blot and quantitative real-time PCR analysis were used to detect the expression of relative molecules. Immunofluorescence staining analysis were also utilized to measure the protein level of CD44. We found that CSC markers, including CD44, CD133, ALDH1-A1, OCT-4 and Nanog, were obviously highly expressed in cell spheres. Moreover, we observed that curcumin reduced the cell spheres formation, decreased the expression of CSC markers, suppressed cell proliferation and induced cell apoptosis. We also found that curcumin inhibited the activation of Shh pathway, while the inhibitory effects of curcumin on BCSCs could be weakened by upregulation of Sonic Hedgehog (Shh) pathway. Altogether, these data suggested that curcumin inhibited the activities of BCSCs through suppressing Shh pathway, which might be an effective chemopreventive agent for bladder cancer intervention.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

Over-expression of PPARα in obese mice adipose tissue improves insulin sensitivity

Peroxisome proliferator-activated receptor α (PPARα) is important in the regulation of lipid metabolism and expressed at high levels in the liver. Although PPARα is also expressed in adipose tissue, little is known about the relationship between its activation and the regulation of glucose metabolism. In this study, we developed adipose tissue specific PPARα over-expression (OE) mice. Metabolomics and insulin tolerance tests showed that OE induces branched-chain amino acid (BCAA) profile and improvement of insulin sensitivity. Furthermore, LC-MS and PCR analyses revealed that OE changes free fatty acid (FFA) profile and reduces obesity-induced inflammation. These findings suggested that PPARα activation in adipose tissue contributes to the improvement of glucose metabolism disorders via the enhancement of BCAA and FFA metabolism.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

miR-21 modulates prostaglandin signaling and promotes gastric tumorigenesis by targeting 15-PGDH

Highlights: • miR-21 is significantly upregulated in gastric cancer. • Up-regulated miR-21 promotes cell proliferation and viability. • miR-21 could target 15-PGDH. • PGE{sub 2}/PI3K/Akt/Wnt/β-catenin axis might be a novel pathway for gastric cancer treatment. miR-21 is an abundantly expressed miRNA in mammalian cells, and evolutionarily conserved across a wide range of vertebrate species. The previous study found that miR-21 is significantly upregulated in gastric cancer. However, the detail mechanisms remain to be largely unknown. In current study, quantitative real-time PCR was applied to examine the expression of miR-21 in gastric cancer tissue and cell lines. The roles of miR-21 in cell proliferation and cell cycle were analyzed by cck8 cell viability assays, flow cytometry cell cycle assays and clone formation assays. As to detail mechanisms, we investigate the relationship between miR-21 and 15-PGDH in gastric cell lines, AGS and BGC-823 treated with In-miR-21, and found that miR-21 is negatively correlated with 15-PGDH. The reduced 15-PGDH may result in PGE2 accumulation which sustains carcinogenesis and tumor progression. We further found that miR-21 exert its oncogenic role through PGE{sub 2}/PI3K/Akt/Wnt/β-catenin axis in gastric cell proliferation. In conclusion, our findings enlarged our knowledge in the roles of miR-21 in the progression of gastric cancer.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Long non-coding RNA-422 acts as a tumor suppressor in colorectal cancer

Highlights: • LncRNA-422 in CRC tissues was significantly lower than that in adjacent tissues. • The expression of lncRNA-422 was correlated with TNM stage of CRC. • LncRNA-422 inhibits the proliferation, migration and invasion of CRC cells. • LncRNA-422 acts as a tumor suppressor by PI3K/AKT/mTOR pathway in CRC. Colorectal cancer (CRC) is one of the most frequent cancer in numerous of countries worldwidely. The initiation and progression of CRC is an extremely complex process, and have been suggested a correlation with Long non-coding RNAs (lncRNAs). Our results showed that lncRNA-422(ENST00000415820) significantly downregulated in the tissues and serum of CRC patients, and is closely associated with the poor prognosis. Then gain or loss of lncRNA-422 models in SW480 and SW620cells were established. The results showed that lncRNA-422 overexpression inhibited cell proliferation, migration, and invasion. Knockdown of lncRNA-422 promoted tumorigensis. Western blot and qRT-PCR were performed to examine the activity of the PI3K/AKT/mTOR pathway in CRC cells after alternation of lncRNA-422. Results showed that lncRNA-422 acts as a tumor suppressor by PI3K/AKT/mTOR pathway in CRC.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

Overexpression of DBC1, correlated with poor prognosis, is a potential therapeutic target for hepatocellular carcinoma

Highlights: • DBC1 was upregulated in HCC tissues and cell lines. • Overexpression of DBC1 was correlated with clinicopathological parameters in HCC. • Overexpression of DBC1 predicts poor disease-free survival (DFS) and overall survival (OS) in HCC. • Knockdown of DBC1 inhibits HCC cell lines proliferation. Deleted in Breast Cancer 1 (DBC1) is a regulatory protein involved in cell metabolism and cancer progression. Nevertheless, the expression and prognostic values of DBC1 in hepatocellular carcinoma (HCC) are still not well understood. The following study investigated the clinical significance and biological function of DBC1 in HCC. Briefly, overexpression of DBC1 at transcriptional and translational levels in human HCC tissues compared to adjacent normal tissues was observed using quantitative real-time polymerase chain reaction (qRT-PCR), western blot (WB) and immunohistochemistry (IHC) approach. Furthermore, upregulated DBC1 was significantly correlated with tumor size (p = 0.005), N stage (p = 0.016), M stage (p = 0.011), tumor differentiation (p in vitro. To conclude, these findings demonstrated that DBC1 was essential in tumorigenesis and proliferation. Moreover, it was identified as a potential therapeutic target for HCC.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

p.Q511L mutation of HNF1α in hepatocellular carcinoma suppresses the transcriptional activity and the anti-tumor effect of HNF1α

Highlights: • A novel somatic mutation p.Q511L of HNF1α was detected in a HCC tissue. • p.Q511L mutation suppressed the transcription activity of HNF1α in HCC cells. • p.Q511L mutation disturbed the subcellular localization of HNF1α. • p.Q511L mutation reduced the inhibitory effect of HNF1α on malignancy of HCC cells. Hepatocyte nuclear factor 1α (HNF1α) is a liver-enriched transcription factor that regulates many aspects of hepatocyte functions. Our previous studies have demonstrated that HNF1α has potent therapeutic effects on hepatocellular carcinoma (HCC). Mutations in HNF1α gene are frequently associated with maturity-onset diabetes of the young type 3 (MODY3) and hepatocellular adenomas. However, the association of HNF1α mutation and HCC remains elusive. In this study, the point mutation of HNF1α gene with c.A1532 > T/p.Q511L was identified in an HCC patient by exon-capture high-throughput sequencing. Mutation of c.A1532 > T/p.Q511L in HNF1α gene was only detected in the tumor tissue but not in the adjacent non-tumorous liver tissue of the patient. Luciferase reporter assay and real-time PCR revealed that mutation of p.Q511L reduced the transcriptional activity of HNF1α. Immunofluorescence staining and subcellular fraction analysis revealed that mutation of p.Q511L disturbed the intracellular localization of HNF1α in HCC cells. Moreover, the inhibitory effect of HNF1α on the proliferation, migration and invasion in HCC cells was also partially abolished by the mutation of p.Q511L. Our data suggested that the missense mutation of HNF1α (p.Q511L) may associate with the progression of HCC.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Salvianolic acid B inhibits myofibroblast transdifferentiation in experimental pulmonary fibrosis via the up-regulation of Nrf2

Highlights: • SalB inhibits TGF-β1-induced myofibroblast transdifferentiation in MRC-5 cells. • SalB ameliorates bleomycin-induced pulmonary fibrosis in rats. • SalB attenuates oxidative stress during experimental pulmonary fibrogenesis. • Nrf2 protein is absent in IPF fibroblasts and fibroblastic foci lesions. • SalB up-regulates the expression of Nrf2 in fibrotic fibroblasts in vitro and vivo. Salvianolic acid B (SalB) is one of the most bioactive components extracted from Salvia miltiorrhiza, and its antioxidant capacity corresponds with its protective effects against cell injury from oxidative stress. The aim of the present study was to evaluate the effect of SalB on experimental pulmonary fibrosis and its ability to ameliorate the oxidative/antioxidative imbalance during fibrosis pathogenesis. The anti-fibrotic activity of SalB was first confirmed in Transforming growth factor β1(TGF-β1)-stimulated MRC-5 cells. The protection of SalB against oxidative stress during fibrogenesis in vitro was verified by detecting ROS production, the levels of glutathione (GSH) and malondialdehyde (MDA). The Western blot and PCR results indicated that SalB could up-regulate nuclear factor erythroid-derived 2-like 2 (Nrf2) at both the protein and mRNA levels and induce Nrf2 nuclear translocation in vitro, which may be the mechanism underlying the anti-fibrotic capacity of SalB. Furthermore, the anti-fibrotic and antioxidant capacities of SalB in vivo were confirmed in rats with BLM-induced pulmonary fibrosis. The immunohistochemistry results showed that Nrf2 was absent in fibroblastic foci (FF) areas, while the SalB treatment could increase the expression of Nrf2 in lung tissues, especially in FF areas.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Secreted glycoprotein BmApoD1 plays a critical role in anti-oxidation and anti-apoptosis in Bombyx mori

Highlights: • Three apolipoprotein D homologs were identified from the genome of lepidopteran insect. • N-linked glycans were essential to the secretion of BmApoD1. • BmApoD1 transcription was up-regulated after oxidant or starvation treatment in larvae. • BmApoD1 can protect cells from oxidative stress and reduce actinomycin D-induced cell apoptosis. Recent studies highlighted that apolipoprotein D (ApoD) and its homologs exert neuroprotective and antioxidant functions in mammals and Drosophila. Unlike mammals and Drosophila, lepidopteran insects possess three distinct ApoD homologs. However, few information on their functions in lepidopteran insects are available. In this study, we investigated the protective potential of a novel ApoD homolog, BmApoD1, in Bombyx mori. Quantitative PCR analyses demonstrated that BmApoD1 is extensively expressed at low levels during the larval stage but abundantly expressed in the testis during the pupal and adult stages. Tryptophan fluorescence titration demonstrated that recombinant BmApoD1 protein can bind retinoic acid and ergosterol. In addition, we provided evidence that N-linked glycans of BmApoD1 are essential to BmApoD1 secretion, and three residues, namely, Asp69, Asp104, and Asp196, are the glycosylation sites of BmApoD1. Furthermore, we showed that BmApoD1 is significantly up-regulated in the larvae after oxidant or starvation treatment. The recombinant BmApoD1 protein can protect cells from oxidative stress induced by H{sub 2}O{sub 2} and reduce actinomycin D-induced cell apoptosis. These observations, together with the transcriptional up-regulation of BmApoD1 in several tissues upon oxidative insult, identify BmApoD1 as a potent antioxidant. Our results demonstrate that BmApoD1 is critical for metabolic adaptation of B. mori to environmental challenges.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Monoterpenoid perillyl alcohol impairs metabolic flexibility of Candida albicans by inhibiting glyoxylate cycle

Highlights: • PA is efficient inhibitor of glyoxylate cycle in Candida albicans which is absent in humans. • PA has negligible hemolytic activity making it viable drug candidate. • PA showed enhanced survival of C. albicans in nematode model, C. elegans demonstrating its in-vivo efficacy. The metabolic pathway such as glyoxylate cycle (GC) enables Candida albicans, to survive under glucose deficient conditions prevalent in the hostile niche. Thus its key enzymes (Isocitrate lyase; ICL and malate synthase; MLS) represent attractive targets against C. albicans. We have previously reported the antifungal potential of a natural monoterpenoid perillyl alcohol (PA). The present study uncovers additional role of PA as a potent GC inhibitor. We explored that PA phenocopied ICL1 deletion mutant and were hypersensitive under low carbon utilizing conditions. The effect of PA on GC was substantiated by molecular docking analyses, which reveals the in-silico binding affinity of PA with ICL and MLS and explored that PA binds to the active sites of both proteins with better binding energy in comparison to their known inhibitors 3-nitropropionate and bromopyruvate respectively. Enzyme kinetics by Lineweaver-Burk plot unravels that PA inhibits ICL and MLS enzymes in competitive and non-competitive manner respectively. Moreover, semi-quantitative RT-PCR indicated that PA inhibits ICL1 and MLS1 mRNA expressions. Lastly, we demonstrated the antifungal efficacy of PA by enhanced survival of Caenorhabditis elegans model and less hemolytic activity (10.6%) on human blood cells. Further studies are warranted for PA to be considered as viable drug candidate.

60 APPLIED LIFE SCIENCES↗