Differences of protein expression of renal cell carcinoma (RCC) by tumor location and treatment.

e15556 Background: Resistance to targeted therapy in RCC is primarily related to tumor and environmental changes. We assessed the difference of morphology and protein analyte expression by metastatic site and treatment types. Methods: Archival tissues from primary and metastatic RCC biopsies were studied. Immunohistochemical probes included antibodies for the protein analytes: phospholipase D2; fatty acid synthase; phosphorylated (p)-Akt (Ser 473); p-mTOR [Ser 2448]); p-p70S6K (Thr 389); p-extracellular signal-regulated kinase (ERK) 1/2 (Thr 202/Tyr 204); p-signal transducer and activator of transcription (STAT)3 (Tyr 705); vascular endothelial growth factor (VEGF)-A; stem cell markers CD44 and CD56; and intratumoral FOXP3 vs. CD8 lymphocytes. Results: A total of 66 patients with available tissue biopsies were assessed. Fourteen patients had additional biopsy at progression. Morphoproteomics was assessed on a total of 81 specimens. The nuclear distribution of phospholipase D2 was statistically significant different by biopsy site (p=0.0046) with highest expression in bone and lung. Similarly, the cytoplasmic expression fatty acid synthase was significantly different by biopsy site (p=0.0167) with highest expression in lymph node and adrenal. More importantly, nuclear mTOR activity was different by tumor biopsy site (p=0.001) with predominant expression in lung, lymph nodes and brain. Compared to mTORC1, which was more expressed in primary tumors, mTORC2 is more expressed in lung and bone metastasis. None of the other protein analytes expression was different based on tumor location. Further analysis on patients with multiple biopsies showed that the plasmalemmal CD56 expression increased with therapy but none of the other protein expression. Conclusions: This analysis emphasized the role of the microenvironment in tumor biology. Prospective treatment trial is underway based on the different molecular fingerprints.

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Effect of Breed on Transcriptional and Protein Expression of Lipogenic Enzymes in Tail and Subcutaneous Adipose Tissue from Two Grazing Breeds of Lambs

Animals ◽

10.3390/ani9020064 ◽

2019 ◽

Vol 9 (2) ◽

pp. 64

Author(s):

María Gallardo ◽

Luis Arias-Darraz ◽

Juan Cárcamo

Keyword(s):

Fatty Acid ◽

Protein Expression ◽

Mrna Expression ◽

Fatty Acid Synthase ◽

Regulatory Element ◽

Subcutaneous Fat ◽

Human Consumption ◽

Meat Industry ◽

Lipogenic Enzymes ◽

Expression Levels

This experiment was carried out to determine the effect of breed on mRNA and protein expression levels of lipogenic enzymes acetyl-CoA carboxylase α (ACC), fatty acid synthase (FAS), stearoyl-CoA desaturase 1 (SCD1) plus sterol regulatory element binding transcription factor 1c (SREBP1c) in the subcutaneous fat (SCF) from the back of the animal, and tail fat (TF) of both Chilota and Suffolk Down lambs grazing Calafatal. Eight Chilota and six Suffolk Down 2-month-old male lambs were allocated to graze a “Calafatal”, a typical secondary succession of Chiloé Archipelago, Chile. After 62 d, lambs were slaughtered according to Chile’s meat industry standards. Fatty acid profile, RT-qPCR, and Western blot analyses from SCF and TF samples were performed. Although the mRNA expression levels of ACC, FAS, SCD1 and SREBP1c in SCF did not differ significantly between breeds (p > 0.05), a trend to higher mRNA expression of FAS and SREBP1c in TF from Chilota lambs was observed (p = 0.06). On the other hand, FAS levels in SCF were higher in Chilota than in Suffolk Down lambs (p < 0.02), although Suffolk Down showed higher fat contents and saturated fatty acid (SFA) proportions than Chilota lambs (p < 0.01). The FAS protein expression in TF was similar in both breeds (p > 0.05). Although the fat content was higher in Suffolk Down than in Chilota lambs (p < 0.01), the SFA proportions were similar in both breeds. Finally, it can be concluded that although mRNA expression of enzymes was similar in both breeds, there were differences in some protein levels in the SCF, partially related with the fatty acid profiles, thus affecting the selection of lamb breed either for human consumption or experimental purposes.

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Polyphenol-Rich Extracts from Toona sinensis Bark and Fruit Ameliorate Free Fatty Acid-Induced Lipogenesis through AMPK and LC3 Pathways

Journal of Clinical Medicine ◽

10.3390/jcm8101664 ◽

2019 ◽

Vol 8 (10) ◽

pp. 1664 ◽

Cited By ~ 3

Author(s):

Yung-Chia Chen ◽

Hsin-Ju Chen ◽

Bu-Miin Huang ◽

Yu-Chi Chen ◽

Chi-Fen Chang

Keyword(s):

Fatty Acid ◽

Liver Disease ◽

Free Fatty Acid ◽

Protein Expression ◽

Lipid Accumulation ◽

Fatty Acid Synthase ◽

Regulatory Element ◽

Autophagic Flux ◽

Alcoholic Fatty Liver ◽

Toona Sinensis

Non-alcoholic fatty liver disease (NAFLD) is a chronic liver disease found worldwide. The present study aimed to evaluate the mechanisms of inhibiting lipid accumulation in free fatty acid (FFA)-treated HepG2 cells caused by bark and fruit extracts of Toona sinensis (TSB and TSF). FFA induced lipid and triglyceride (TG) accumulation, which was attenuated by TSB and TSF. TSB and/or TSF promoted phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-coA carboxylase and peroxisome proliferator-activated receptor alpha upregulation. Furthermore, TSB and TSF suppressed FFA-induced liver X receptor, sterol regulatory element-binding transcription protein 1, fatty acid synthase, and stearoyl-CoA desaturase 1 protein expression. Moreover, TSB and/or TSF induced phosphorylation of Unc-51 like autophagy-activating kinase and microtubule-associated protein 1A/1B-light chain 3 expressions. Therefore, TSB and TSF relieve lipid accumulation by attenuating lipogenic protein expression, activating the AMPK pathway, and upregulating the autophagic flux to enhance lipid metabolism. Moreover, TSB and TSF reduced TG contents, implying the therapeutic use of TSB and TSF in NAFLD.

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Proteomic Analysis of Waldenstrom Macroglobulinemia (WM) and IgM Monoclonal Gammopathy of Undetermined Significance (IgM-MGUS) Identifies Potential Targets for Drug Therapy.

Blood ◽

10.1182/blood.v104.11.787.787 ◽

2004 ◽

Vol 104 (11) ◽

pp. 787-787

Author(s):

Irene M. Ghobrial ◽

Alexey A. Leontocich ◽

Jeanette E. Eckel ◽

Michael Timm ◽

Steve Ziesmer ◽

...

Keyword(s):

Bone Marrow ◽

Fatty Acid ◽

Protein Expression ◽

Disease Progression ◽

Normal Control ◽

Fatty Acid Synthase ◽

Protein Microarrays ◽

Small Gtpases ◽

Normal Donor ◽

Cell Extracts

Abstract The objective of this study was to analyze molecular changes that occur at the protein level in WM and IgM-MGUS in order to identify proteins dysregulated at disease progression and design new therapies. We employed antibody protein microarrays (BD Clonetech, CA) to measure changes in the patterns of protein expression between WM, IgM-MGUS and normal lymphoplasmacytic cells as control. The antibody array is a novel technique that assays protein differences directly by hybridizing fluorescently labeled protein mixtures from cell extracts onto a glass slide spotted with 512 different monoclonal antibodies specific for human proteins. CD 19+ and CD138+ purified lymphoplasmacytic cells were obtained from cryopreserved bone marrow samples of 5 newly diagnosed patients with WM and 5 patients with IgM-MGUS. Control plasma cells were obtained from 3 pooled CD19+ and CD138+ purified normal donor bone marrow cells. To assess differential expression, the mean of the ratios of Cy5/Cy3 for each sample were analyzed using the Clontech software to calculate an internally normalized ratio. The normalized data were analyzed by the Genespring software and changes of protein expression ≥2 fold in 60% of the samples as compared to normal control were identified in both WM and MGUS groups. There were 20 proteins upregulated in WM and 21 proteins in MGUS as compared to normal control cells These included proteins from the small GTPases such as the Ras, Rho and Ran families (Rab 4, MONA, RCC1, and CDC42GAP), Kinases/signaling proteins (ROK alpha, p62 dok, JAK, 14-3-3e), cyclin dependent kinase (CDK2), fatty acid synthase (HDAC3), NFkB pathway (IKKalpha, IKBe), and histone deacetylase. The WM and IgM MGUS differed in only 3 proteins using the 2-fold threshold. The WM samples showed overexpression of the heat shock protein Hsp90, the Ras family protein CDC25C, and the chemotaxis protein p43/EMAPII as compared to the MGUS samples. This is the first proteomic study of WM and MGUS patients. It identifies novel proteins dysregulated in WM that may serve in future clinical trials as targets for therapy such as Hsp90, NFkB, HDAC and fatty acid synthase. In addition, it identifies 3 proteins that identify disease progression from MGUS to WM, which could be useful for monitoring MGUS patients and designing targeted therapies to prevent progression to WM.

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Proteomic Analysis of Targeted Therapeutic Agents in Multiple Myeloma (MM).

Blood ◽

10.1182/blood.v104.11.4299.4299 ◽

2004 ◽

Vol 104 (11) ◽

pp. 4299-4299

Author(s):

Irene M. Ghobrial ◽

Jeanette E. Eckel ◽

Antje Hoering ◽

Michael Timm ◽

Alexey A. Leontovich ◽

...

Keyword(s):

Clinical Trials ◽

Fatty Acid ◽

Protein Expression ◽

Rational Design ◽

Fatty Acid Synthase ◽

Fatty Acid Synthetase ◽

Pi3k Pathway ◽

Therapeutic Agents ◽

Protein Arrays ◽

Cell Extracts

Abstract The objective of this study was to analyze protein changes that occur a the molecular level with novel therapeutic agents used in MM in order to identify potential combinations for clinical trials. We employed antibody protein microarrays (BD Clonetech, CA) to measure changes in the patterns of protein expression in Kas 6/1 MM cell line after treatment with bortezomib, dexamethasone, CC-5013, and 17-AAG. The antibody array is a new technique that assays protein differences directly by hybridizing fluorescently labeled protein mixtures from cell extracts onto glass slides spotted with 512 different monoclonal antibodies specific for human proteins. Induction of apoptosis was assessed by Annexin/Propidium iodide FACS analysis at 24 and 48 hours using different concentrations of the inhibitors. Protein arrays were performed at concentrations and treatment times that did not induce more than 25% apoptosis to ensure adequate analysis of early changes in signaling pathways. Bortezomib 5nM at 10 hours, dexamethasone 100mM, CC-5013 250nM, and 17-AAG 1mM at 24 hours were used. Control cells were treated with vehicles ETOH or DMSO at the same concentrations as the inhibitors. To assess differential protein expression, the mean of the Cy5/Cy3 ratios for each sample and its control were analyzed using the Clontech software and an internally normalized ratio (INR) was calculated. INR values >1.3 or <0.77 were considered as valid changes in protein abundance for this study. All inhibitors induced the upregulation of apoptosis proteins including BAK, BIK, SMAC/DIABLO, and caspase 14. Several proteins were inhibited by all 4 inhibitors indicating common pathways affected by these agents. These included members of the PI3K pathway (PI3K-p85a, p70S6kinase, and cyclin dependent kinases), MAPKinases (ERK1 and 2), antiapoptotic protein Bclx, heat shock proteins Hsp70 and HSP60, and ubiqutin/NFkB pathway (Ubch6, IKKalpha, and IKBe). These results indicate that inhibitors of the PI3K pathway such as mTOR inhibitors, IKK inhibitors or MAPkinase inhibitors may be used in the future in combination with bortezomib, dexamethasone, CC-5013 or 17-AAG. Fatty acid synthetase was downregulated by bortezomib and dexamethasone indicating potential use of fatty acid synthase inhibitors in combination with bortezomib or dexamethasone in future clinical trials. These results suggest that antibody protein arrays may be used in the rational design of combinations of targeted therapies in MM.

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Expression of Fatty Acid Synthase Depends on NAC1 and Is Associated with Recurrent Ovarian Serous Carcinomas

Journal of Oncology ◽

10.1155/2010/285191 ◽

2010 ◽

Vol 2010 ◽

pp. 1-12 ◽

Cited By ~ 29

Author(s):

Stefanie M. Ueda ◽

Kai Lee Yap ◽

Ben Davidson ◽

Yuan Tian ◽

Vivek Murthy ◽

...

Keyword(s):

Ovarian Cancer ◽

Fatty Acid ◽

Cancer Cells ◽

Fatty Acid Synthase ◽

Dominant Negative ◽

Serous Carcinoma ◽

Ovarian Cancer Cells ◽

Primary Tumors ◽

Ovarian Serous Carcinoma ◽

Skov3 Cells

Our previous reports demonstrated that NAC1, a BTB/POZ domain-containing nuclear protein, upregulates in recurrent ovarian serous carcinoma and participates in developing drug resistance in cancer cells. The current study applies quantitative proteomics to identify the proteins controlled by NAC1 by comparing the proteomes of SKOV3 cells with and without expression of a dominant negative NAC1 construct, N130. From the proteins that are downregulated by N130 (upregulated by NAC1), we chose to further characterize fatty acid synthase (FASN). Similar to change in protein level, the FASN transcript level in SKOV3 cells was significantly reduced by N130 induction or by NAC1 knockdown. Immunohistochemistry showed that NAC1 and FASN immunointensities in ovarian serous carcinoma tissues had a highly significant correlation (P<.0001). Moreover, we found that recurrent serous carcinomas exhibited higher FASN immunointensities than their matched primary tumors (P<.001). Multivariate analysis showed that an FASN staining score of >1 in serous carcinomas was associated with a worse overall survival time (P<.01). Finally, C93, a new FASN inhibitor, induced massive apoptosis in carboplatin/paclitaxel resistant ovarian cancer cells. In conclusion, we show that NAC1 is essential for FASN expression in ovarian serous carcinomas and the expression of FASN significantly correlates with tumor recurrence and disease aggressiveness. The dependence of drug resistant tumor cells on FASN suggests a potential application of FASN-based therapeutics for recurrent ovarian cancer patients.

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Synthesis of new C75 derivatives, Fatty Acid Synthase inhibitors with cytotoxic properties

Bionatura ◽

10.21931/rb/cs/2019.02.01.9 ◽

2019 ◽

Vol 02 (Bionatura Conference Serie) ◽

Cited By ~ 1

Author(s):

Kamil Makowski ◽

Javier Ariza ◽

Jordi Garcia ◽

Laura Herrero ◽

Marta López ◽

...

Keyword(s):

Fatty Acid ◽

Fatty Acid Synthase ◽

De Novo ◽

Human Cancer ◽

Acid Synthesis ◽

The Other ◽

Aliphatic Chain ◽

Key Enzyme ◽

Cancer Tissues ◽

Derivatives Of

The fatty-acid synthase (FAS) is the key enzyme that executes de novo fatty acid synthesis and is overexpressed in some human cancer tissues. C75 is a synthetic FAS inhibitor, which produces the death of cancer cells. In this work, we synthesized three derivatives of C75, two homologs with 1 and 2 carbons more in the beta position of the lactone and the other one with a change in the elongation of the aliphatic chain in gamma position from 8 to 15 carbons. The results from FAS inhibition and cytotoxic properties suggest that the performed changes in the derivatives decrease their activity.

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Resistant Maltodextrin Ameliorates Altered Hepatic Lipid Homeostasis via Activation of AMP-Activated Protein Kinase in a High-Fat Diet-Fed Rat Model

Nutrients ◽

10.3390/nu11020291 ◽

2019 ◽

Vol 11 (2) ◽

pp. 291 ◽

Cited By ~ 6

Author(s):

Shing-Hwa Liu ◽

Chen-Yuan Chiu ◽

Lin-Hui Huang ◽

Meng-Tsan Chiang

Keyword(s):

Fatty Acid ◽

Protein Kinase ◽

Protein Expression ◽

Fatty Liver ◽

Fatty Acid Synthase ◽

Blood Cholesterol ◽

Peroxisome Proliferator ◽

High Fat ◽

Resistant Maltodextrin ◽

Amp Activated Protein Kinase

Many studies have shown that resistant maltodextrin (RMD) possesses blood cholesterol lowering and anti-obesity effects. In order to investigate the effect of RMD on lipid metabolism in the liver, rats were fed with a high-fat (HF) diet for 7 weeks to induce hyperlipidemia and fatty liver. Normal control rats were fed with a normal diet. HF-diet-fed rats were treated with 5% RMD for 8 weeks. The results showed that the increased plasma aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities, the increased hepatic triglyceride and total cholesterol levels, and fatty liver in HF-diet-fed rats were significantly decreased after supplementation with RMD. Supplementation with RMD significantly (1) induced AMP-activated protein kinase (AMPK) phosphorylation; (2) inhibited the activities of acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), and HMG-CoA reductase (HMGCR); (3) suppressed the protein expression of peroxisome proliferator activated receptor (PPAR)-γ; (4) increased β-oxidation of fatty acids by increasing the protein expression carnitine palmitoyl transferase 1α (CPT-1α) in the livers of HF-diet-fed rats. Taken together, supplementation of RMD was capable of inhibiting lipogenic enzyme activities and inducing fatty acid β-oxidation through increasing AMPK activation, thereby reducing lipid accumulation in the liver.

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Analyzing effects of extra-virgin olive oil polyphenols on breast cancer-associated fatty acid synthase protein expression using reverse-phase protein microarrays

International Journal of Molecular Medicine ◽

10.3892/ijmm_00000039 ◽

1998 ◽

Vol 22 (4) ◽

Cited By ~ 6

Author(s):

Menendez

Keyword(s):

Breast Cancer ◽

Fatty Acid ◽

Protein Expression ◽

Olive Oil ◽

Fatty Acid Synthase ◽

Virgin Olive Oil ◽

Extra Virgin Olive Oil ◽

Protein Microarrays ◽

Reverse Phase ◽

Phase Protein

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FAS inhibitor cerulenin reduces food intake and melanocortin receptor gene expression without modulating the other (an)orexigenic neuropeptides in chickens

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.00899.2005 ◽

2006 ◽

Vol 291 (1) ◽

pp. R138-R147 ◽

Cited By ~ 27

Author(s):

Sami Dridi ◽

Cedric Ververken ◽

F. Bradley Hillgartner ◽

Arckens Lutgarde ◽

Estel Van der Gucht ◽

...

Keyword(s):

Gene Expression ◽

Fatty Acid ◽

Food Intake ◽

Broiler Chickens ◽

Fatty Acid Synthase ◽

Ingestive Behavior ◽

The Other ◽

Melanocortin Receptor ◽

Synthetic Analog ◽

Mrna Levels

Cerulenin, a natural fatty acid synthase (FAS) inhibitor, and its synthetic analog C75 are hypothesized to alter the metabolism of neurons in the hypothalamus that regulate ingestive behavior to cause a profound decrease of food intake and an increase in metabolic rate, leading to body weight loss. The bulk of data exclusively originates from mammals (rodents); however, such effects are currently lacking in nonmammalian species. We have, therefore, addressed this issue in broiler chickens because this species is selected for high growth rate and high food intake and is prone to obesity. First, we demonstrate that FAS messenger and protein are expressed in the hypothalamus of chickens. FAS immunoreactivity was detected in a number of brain regions, including the nucleus paraventricularis magnocellularis and the nucleus infundibuli hypothalami, the avian equivalent of the mammalian arcuate nucleus, suggesting that FAS may be involved in the regulation of food intake. Second, we show that hypothalamic FAS gene expression was significantly ( P < 0.05) decreased by overnight fasting similar to that in liver, indicating that hypothalamic FAS gene is regulated by energy status in chickens. Finally, to investigate the physiological consequences of in vivo inhibition of fatty acid synthesis on food intake, we administered cerulenin by intravenous injections (15 mg/kg) to 2-wk-old broiler chickens. Cerulenin administration significantly reduced food intake by 23 to 34% ( P < 0.05 to P < 0.0001) and downregulated FAS and melanocortin receptors 1, 4, and 5 gene expression ( P < 0.05). However, the known orexigenic (neuropeptide Y, agouti gene-related peptide, orexin, and orexin receptor) and anorexigenic (pro-opiomelanocortin and corticotropin-releasing hormone) neuropeptide mRNA levels remained unchanged after cerulenin treatment. These results suggest that the catabolic effect of cerulenin in chickens may be mediated through the melanocortin system rather than the other neuropeptides known to be involved in food intake regulation.

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