19 research outputs found
TGFβ Drives Metabolic Perturbations during Epithelial Mesenchymal Transition in Pancreatic Cancer: TGFβ Induced EMT in PDAC
TGF beta; Pancreatic cancer; Tumor microenvironmentTGF beta; Cancer de pancreas; Microambiente tumoralTGF beta; Càncer de pàncrees; Microambient tumoralPancreatic ductal adenocarcinoma (PDAC) is a highly lethal malignancy wherein a majority of patients present metastatic disease at diagnosis. Although the role of epithelial to mesenchymal transition (EMT), mediated by transforming growth factor beta (TGFβ), in imparting an aggressive phenotype to PDAC is well documented, the underlying biochemical pathway perturbations driving this behaviour have not been elucidated. We used high-resolution mass spectrometry (HRMS) based molecular phenotyping approach in order to delineate metabolic changes concomitant to TGFβ-induced EMT in pancreatic cancer cells. Strikingly, we observed robust changes in amino acid and energy metabolism that may contribute to tumor invasion and metastasis. Somewhat unexpectedly, TGFβ treatment resulted in an increase in intracellular levels of retinoic acid (RA) that in turn resulted in increased levels of extracellular matrix (ECM) proteins including fibronectin (FN) and collagen (COL1). These findings were further validated in plasma samples obtained from patients with resectable pancreatic cancer. Taken together, these observations provide novel insights into small molecule dysregulation that triggers a molecular cascade resulting in increased EMT-like changes in pancreatic cancer cells, a paradigm that can be potentially targeted for better clinical outcomes.This study was supported by American Cancer Society (IRG-92-152-17 award number AWD4470404), Georgetown Lombardi Comprehensive Cancer Center Support Grant Developmental Funds and Ruesch Foundation to K.U. and A.K.C
LC-MS/MS profile of an active pharmaceutical ingredient and its impurities in commercial preparation
An Optimized Method for LC–MS-Based Quantification of Endogenous Organic Acids: Metabolic Perturbations in Pancreatic Cancer
Accurate and reliable quantification of organic acids with carboxylic acid functional groups in complex biological samples remains a major analytical challenge in clinical chemistry. Issues such as spontaneous decarboxylation during ionization, poor chromatographic resolution, and retention on a reverse-phase column hinder sensitivity, specificity, and reproducibility in multiple-reaction monitoring (MRM)-based LC–MS assays. We report a targeted metabolomics method using phenylenediamine derivatization for quantifying carboxylic acid-containing metabolites (CCMs). This method achieves accurate and sensitive quantification in various biological matrices, with recovery rates from 90% to 105% and CVs ≤ 10%. It shows linearity from 0.1 ng/mL to 10 µg/mL with linear regression coefficients of 0.99 and LODs as low as 0.01 ng/mL. The library included a wide variety of structurally variant CCMs such as amino acids/conjugates, short- to medium-chain organic acids, di/tri-carboxylic acids/conjugates, fatty acids, and some ring-containing CCMs. Comparing CCM profiles of pancreatic cancer cells to normal pancreatic cells identified potential biomarkers and their correlation with key metabolic pathways. This method enables sensitive, specific, and high-throughput quantification of CCMs from small samples, supporting a wide range of applications in basic, clinical, and translational research
Identifying inflammation-related targets of natural lactones using network pharmacology, molecular modeling and <i>in vitro</i> approaches
Natural lactones have been used in traditional and folklore medicine for centuries owing to their anti-inflammatory properties. The study uses a multifaceted approach to identify lead anti-inflammatory lactones from the SISTEMATX natural products database. The study analyzed the natural lactone database, revealing 18 lactones linked to inflammation targets. The primary targets were PTGES, PTGS1, COX-2, ALOX5 and IL1B. STX 12273 was the best hit, with the lowest binding energy and potential for inhibiting the COX-2 enzyme. The study suggested natural lactone, STX 12273, from the SISTEMATX database with anti-inflammatory potential and postulated its use for inflammation treatment or prevention. Communicated by Ramaswamy H. Sarma</p
Glycyrrhiza glabra extract and quercetin reverses cisplatin resistance in triple-negative MDA-MB-468 breast cancer cells via inhibition of cytochrome P450 1B1 enzyme
© 2017 Elsevier Ltd The development of multi-drug resistance to existing anticancer drugs is one of the major challenges in cancer treatment. The over-expression of cytochrome P450 1B1 enzyme has been reported to cause resistance to cisplatin. With an objective to discover cisplatin-resistance reversal agents, herein, we report the evaluation of Glycyrrhiza glabra (licorice) extracts and its twelve chemical constituents for inhibition of CYP1B1 (and CYP1A1) enzyme in Sacchrosomes and live human cells. The hydroalcoholic extract showed potent inhibition of CYP1B1 in both Sacchrosomes as well as in live cells with IC50 values of 21 and 16 µg/mL, respectively. Amongst the total of 12 constituents tested, quercetin and glabrol showed inhibition of CYP1B1 in live cell assay with IC50 values of 2.2 and 15 µM, respectively. Both these natural products were found to be selective inhibitors of CYP1B1, and does not inhibit CYP2 and CYP3 family of enzymes (IC50 > 20 µM). The hydroalcoholic extract of G. glabra and quercetin (4) showed complete reversal of cisplatin resistance in CYP1B1 overexpressing triple negative MDA-MB-468 breast cancer cells. The selective inhibition of CYP1B1 by quercetin and glabrol over CYP2 and CYP3 family of enzymes was studied by molecular modeling studies
Establishment of LCMS Based Platform for Discovery of Quorum Sensing Inhibitors: Signal Detection in <i>Pseudomonas aeruginosa</i> PAO1
Targeting
the main three networking systems, <i>viz.</i> Las, RhI,
and PQS, <i>via</i> natural quenchers is a new
ray of hope for combating the persistent behavior of <i>Pseudomonas
aeruginosa</i>. In the bacterial chemical vocabulary pyocyanin,
N-AHLs and rhamnolipids are the main keywords, which are responsible
for the social and nomadic behavior of <i>P. aeruginosa</i>. In the present work, LC-MS based real-time qualitative and quantitative
analysis of pyocyanin, green phenazine, N-AHLs, and rhamnolipids were
performed on <i>P. aeruginosa</i> PAO1. The quantitative
analysis indicates that the production of pyocyanin and NHSLs increases
with time while the production of rhamnolipids discontinued after
16 h. This indicates the emergence of persisters in the medium instead
of planktonic cells. Rhamnolipids acting as a surfactant enhances
the motility of the bacterial cells, whereas the pyocyanin is responsible
for the biofilm formation. In a microtiter plate based assay, an effect
of capsaicin and 6-gingerol was recorded. In the presence of capsaicin
and 6-gingerol, a substantial decrease in the production of rhamnolipids,
phenazine, quinolone, and N-AHLs was observed. Most interestingly,
the 6-gingerol treatment led to a drastic decrease of rhamnolipids,
phenazine, quinolone, and N-AHLs versus capsaicin. These studies demonstrate
the effectiveness of the capsaicin and 6-gingerol on Las, PQS, and
Rhl circuits in a bacterium in order to understand the persistent
and social behavior. Here, we are reporting LC-MS/MS based qualitative
and quantitative analysis of QS molecules by taking a low volume of
culture (up to 200 μL). This method can be used as a platform
to screen the new antivirulence agents for fighting the resistant
behavior of <i>P. aeruginosa</i> during biofilm formation
<i>ortho</i>-Amidoalkylation of Phenols via Tandem One-Pot Approach Involving Oxazine Intermediate
A new and efficient method for <i>ortho</i>-amidoalkylation
of phenols via Mannich-type condensation with formaldehyde and lactams
using recyclable solid acid catalyst is described. This is the first
report for <i>ortho</i>-amidoalkylation of phenols by lactams
via Mannich-type condensation. LC-ESI-MS/MS based mechanistic study
revealed that reaction proceeds through <i>o</i>-quinone
methide (<i>o</i>-QM) and an oxazine intermediate via tandem
Knoevenagel condensation, formal [4 + 2]-Diels–Alder cycloaddition
and acid catalyzed oxazine ring-opening
Semisynthesis of Mallotus B from Rottlerin: Evaluation of Cytotoxicity and Apoptosis-Inducing Activity
Mallotus B (<b>2d</b>) is a
prenylated dimeric phloroglucinol
compound isolated from <i>Mallotus philippensis.</i> There
have been no reports on the synthesis or biological activity of this
compound. In the present paper, a semisynthetic preparation of mallotus
B is reported via base-mediated intramolecular rearrangement of rottlerin
(<b>1</b>), which is one of the major constituents of <i>M. philippensis</i>. The homodimer “rottlerone”
was also formed as one of the products of this base-mediated intramolecular
reaction. Rottlerin (<b>1</b>), along with rottlerone (<b>2c</b>) and mallotus B (<b>2d</b>), was evaluated for cytotoxicity
against a panel of cancer cell lines including HEPG2, Colo205, MIAPaCa-2,
PC-3, and HL-60 cells. Mallotus B (<b>2d</b>) displayed cytotoxicity
for MIAPaCa-2 and HL-60 cells with IC<sub>50</sub> values of 9 and
16 μM, respectively. Microscopic studies in HL-60 cells indicated
that mallotus B (<b>2d</b>) induces cell cycle arrest at the
G1 phase and causes defective cell division. It also induces apoptosis,
as evidenced by distinct changes in cell morphology
TGFβ Drives Metabolic Perturbations during Epithelial Mesenchymal Transition in Pancreatic Cancer: TGFβ Induced EMT in PDAC.
Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal malignancy wherein a majority of patients present metastatic disease at diagnosis. Although the role of epithelial to mesenchymal transition (EMT), mediated by transforming growth factor beta (TGFβ), in imparting an aggressive phenotype to PDAC is well documented, the underlying biochemical pathway perturbations driving this behaviour have not been elucidated. We used high-resolution mass spectrometry (HRMS) based molecular phenotyping approach in order to delineate metabolic changes concomitant to TGFβ-induced EMT in pancreatic cancer cells. Strikingly, we observed robust changes in amino acid and energy metabolism that may contribute to tumor invasion and metastasis. Somewhat unexpectedly, TGFβ treatment resulted in an increase in intracellular levels of retinoic acid (RA) that in turn resulted in increased levels of extracellular matrix (ECM) proteins including fibronectin (FN) and collagen (COL1). These findings were further validated in plasma samples obtained from patients with resectable pancreatic cancer. Taken together, these observations provide novel insights into small molecule dysregulation that triggers a molecular cascade resulting in increased EMT-like changes in pancreatic cancer cells, a paradigm that can be potentially targeted for better clinical outcomes