Quantitative Proteomics: Measuring Protein Synthesis Using15N Amino Acid Labeling in Pancreatic Cancer Cells

Gemcitabine is the first-line chemotherapy for pancreatic cancer. To overcome the often-acquired gemcitabine resistance, other drugs are used in combination with gemcitabine. It is well-known that cancer cells reprogram cellular metabolism, coupled with the up-regulation of selective nutrient transporters to feed into the altered metabolic pathways. Our previous studies have demonstrated that the amino acid transporter SLC6A14 is markedly up-regulated in pancreatic cancer and that it is a viable therapeutic target. α-Methyltryptophan (α-MT) is a blocker of SLC6A14 and is effective against pancreatic cancer in vitro and in vivo. In the present study, we tested the hypothesis that α-MT could synergize with gemcitabine in the treatment of pancreatic cancer. We investigated the effects of combination of α-MT and gemcitabine on proliferation, migration, and apoptosis in a human pancreatic cancer cell line, and examined the underlying mechanisms using 1H-NMR-based metabolomic analysis. These studies examined the intracellular metabolite profile and the extracellular metabolite profile separately. Combination of α-MT with gemcitabine elicited marked changes in a wide variety of metabolic pathways, particularly amino acid metabolism with notable alterations in pathways involving tryptophan, branched-chain amino acids, ketone bodies, and membrane phospholipids. The metabolomic profiles of untreated control cells and cells treated with gemcitabine or α-MT were distinctly separable, and the combination regimen showed a certain extent of overlap with the individual α-MT and gemcitabine groups. This represents the first study detailing the metabolomic basis of the anticancer efficacy of gemcitabine, α-MT and their combination.

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Mass Defect-Based N,N-Dimethyl Leucine Labels for Quantitative Proteomics and Amine Metabolomics of Pancreatic Cancer Cells

Analytical Chemistry ◽

10.1021/acs.analchem.6b03482 ◽

2017 ◽

Vol 89 (2) ◽

pp. 1138-1146 ◽

Cited By ~ 27

Author(s):

Ling Hao ◽

Jillian Johnson ◽

Christopher B. Lietz ◽

Amanda Buchberger ◽

Dustin Frost ◽

...

Keyword(s):

Pancreatic Cancer ◽

Cancer Cells ◽

Quantitative Proteomics ◽

Mass Defect ◽

Pancreatic Cancer Cells

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Synthesis of Gemcitabine-Threonine Amide Prodrug Effective on Pancreatic Cancer Cells with Improved Pharmacokinetic Properties

Molecules ◽

10.3390/molecules23102608 ◽

2018 ◽

Vol 23 (10) ◽

pp. 2608 ◽

Cited By ~ 5

Author(s):

Sungwoo Hong ◽

Zhenghuan Fang ◽

Hoi-Yun Jung ◽

Jin-Ha Yoon ◽

Soon-Sun Hong ◽

...

Keyword(s):

Pancreatic Cancer ◽

Amino Acid ◽

Cancer Cells ◽

Systemic Exposure ◽

Amino Acid Transporter ◽

Amide Bond ◽

Amino Acid Transporters ◽

Pancreatic Cancer Cells ◽

Acid Transporter

To investigate the amino acid transporter-based prodrug anticancer strategy further, several amino acid-conjugated amide gemcitabine prodrugs were synthesized to target amino acid transporters in pancreatic cancer cells. The structures of the synthesized amino acid-conjugated prodrugs were confirmed by 1H-NMR and LC-MS. The pancreatic cancer cells, AsPC1, BxPC-3, PANC-1 and MIAPaCa-2, appeared to overexpress the amino acid transporter LAT-1 by conventional RT-PCR. Among the six amino acid derivatives of gemcitabine, threonine derivative of gemcitabine (Gem-Thr) was more effective than free gemcitabine in the pancreatic cancer cells, BxPC-3 and MIAPaCa-2, respectively, in terms of anti-cancer effects. Furthermore, Gem-Thr was metabolically stable in PBS (pH 7.4), rat plasma and liver microsomal fractions. When Gem-Thr was administered to rats at 4 mg/kg i.v., Gem-Thr was found to be successfully converted to gemcitabine via amide bond cleavage. Moreover, the Gem-Thr showed the increased systemic exposure of formed gemcitabine by 1.83-fold, compared to free gemcitabine treatment, due to the significantly decreased total clearance (0.60 vs. 4.23 mL/min/kg), indicating that the amide prodrug approach improves the metabolic stability of gemcitabine in vivo. Taken together, the amino acid transporter-targeting gemcitabine prodrug, Gem-Thr, was found to be effective on pancreatic cancer cells and to offer an efficient potential means of treating pancreatic cancer with significantly better pharmacokinetic characteristics than gemcitabine.

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Inhibition of protein synthesis by imexon reduces HIF-1α expression in normoxic and hypoxic pancreatic cancer cells

Investigational New Drugs ◽

10.1007/s10637-008-9149-9 ◽

2008 ◽

Vol 27 (1) ◽

pp. 89-98 ◽

Cited By ~ 7

Author(s):

Betty K. Samulitis ◽

Terry H. Landowski ◽

Robert T. Dorr

Keyword(s):

Pancreatic Cancer ◽

Protein Synthesis ◽

Cancer Cells ◽

Pancreatic Cancer Cells ◽

Inhibition Of Protein Synthesis

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Abstract 4533: Mechanism of anti-proliferative effects of sanguinarine in pancreatic cancer cells: A label-free quantitative proteomics approach

10.1158/1538-7445.am2014-4533 ◽

2014 ◽

Author(s):

Chandra K. Singh ◽

Satwinderjeet Kaur ◽

Jasmine George ◽

Molly C. Pellitteri-Hahn ◽

Cameron O. Scarlett ◽

...

Keyword(s):

Pancreatic Cancer ◽

Cancer Cells ◽

Quantitative Proteomics ◽

Label Free ◽

Pancreatic Cancer Cells ◽

Proteomics Approach

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Effect of LAT2 on glutamine-dependent mTOR activation to promote glycolysis and chemoresistance in pancreatic cancer.

Journal of Clinical Oncology ◽

10.1200/jco.2019.37.15_suppl.e15759 ◽

2019 ◽

Vol 37 (15_suppl) ◽

pp. e15759-e15759

Author(s):

Jiangdong Qiu ◽

Mengyu Feng ◽

Zhe Cao ◽

Gang Yang ◽

Yueze Liu ◽

...

Keyword(s):

Pancreatic Cancer ◽

Amino Acid ◽

Cancer Cells ◽

Amino Acid Transporter ◽

Glutamine Metabolism ◽

Clinical Value ◽

Pancreatic Cancer Cells ◽

Acid Transporter

e15759 Background: Reprogrammed energy metabolism has become the characteristic of cancer recently. Transporters act as amino acid sensors involved in mTOR recruitment and activation, which is crucial for the growth of both normal and tumor cells. L-type amino acid transporter 2 (LAT2), a Na+ -independent neutral amino acid transporter, is encoded by the SLC7A8 gene and responsible for transporting neutral amino acids, including a mTOR activator, glutamine. LAT2 was reported to be overexpressed in gemcitabine-resistant pancreatic cancer cells. However, the role of LAT2 in chemoresistance in pancreatic cancer remains unclear. Methods: The effects of LAT2 on biological behaviors of pancreatic cancer cells were analyzed. LAT2 and LDHB levels in tissues were detected, and the clinical value was evaluated. Results: We demonstrated that LAT2 played an oncogenic role and decreased the gemcitabine sensitivity of pancreatic cancer cells in vitro and in vivo. Survival analysis indicated that high expression of both LAT2 and LDHB was related to a poor prognosis in patients with pancreatic cancer. Furthermore, we found that LAT2 could promote proliferation, inhibit apoptosis, activate glycolysis and alter glutamine metabolism to activate mTOR in vitro and in vivo. Next, the combination of gemcitabine with an mTOR inhibitor (RAD001) could reverse the decrease in chemosensitivity caused by LAT2 overexpression in pancreatic cancer cells. Mechanistically, LAT2 promoted glycolysis and decreased gemcitabine sensitivity via regulating two glutamine-dependent positive feedback loops (the LAT2/p-mTORSer2448 loop and the glutamine/p-mTORSer2448/glutamine synthetase loop) in pancreatic cancer. Conclusions: Our data indicates that LAT2 functions as an oncogenic protein and could regulate glutamine-dependent mTOR activation to promote glycolysis and decrease gemcitabine sensitivity in pancreatic cancer. The LAT2-mTOR-LDHB pathway might be a promising therapeutic target in pancreatic cancer.

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Albumin Based Nanoparticles for Detection of Pancreatic Cancer Cells

Protein and Peptide Letters ◽

10.2174/0929866526666190119121434 ◽

2019 ◽

Vol 26 (4) ◽

pp. 271-280

Author(s):

Nursenem Karaca ◽

Özlem Biçen Ünlüer

Keyword(s):

Pancreatic Cancer ◽

Amino Acid ◽

Cancer Cells ◽

Cancer Cell ◽

Circulatory System ◽

Research Area ◽

Cross Linking ◽

Pancreatic Cancer Cells ◽

Binding Function

Background:Molecular imaging of cancer cells using effective drug targeting systems are most interested research area in recent years. Albumin protein is a soluble and most abundant protein in circulatory system. It has a ligand-binding function and acts as a transport protein. Researchers are interested in developing albumin based nanostructured specific anti-tumor drugs in cancer therapy. Pancreatic cancer treatment or drug design for targeted pancreatic cancer cell has great importance due to it has a high mortality rate comparing other cancer types.Objective:In this article, our goal is to develop new targeting nanoparticles based on the conjugation of albumin and Hyaluronic Acid (HA) for pancreatic cancer cells.Method:In this article, we proposed a new technique for conjugation of albumin (BSA) and HA in nano formation. Firstly, cationic BSA is synthesized. Then, BSA-HA conjugation is obtained by interacted cationic BSA with 1000 ppm HA. Secondly, nano BSA-HA particles and nano BSA particles were synthesized according to AmiNoAcid Decorated and Light Underpinning Conjugation Approach (ANADOLUCA) method which provides a special cross-linking strategy for biomolecules using ruthenium-based amino acid monomer haptens. After characterization studies, in vitro cytotoxic activity of synthesized nano BSA-HA particles were determined for PANC-1 ATCC® CRL146 cells.Results:According to the data, nano BSA and nano BSA-HA particles synthesized uniquely using special ruthenium-based amino acid decorated cross-linking agent, (MATyr)2-Ru-(MATyr)2.based on ANDOLUCA method. Characterization results showed that there was not any change in protein folding structures during nano formation process. In addition, nano protein particles gained fluorescence feature. When interacting synthesized nano BSA and nano BSA-HA particles with pancreatic cells, it was found that BSA nanoparticles were usually around cells and membranes, but BSA-HA nanoparticles were identified around the cells, in the cytoplasm inside the cell, and next to the cell nucleus. So, nano BSA-HA particles could be used as cancer cell imaging agent for PANC-1 ATCC® CRL146 cells.Conclusion:The satisfactory conclusion of this study is that synthesized nano BSA-HA particles are fundamental materials for targeting pancreatic cancer cells due to HA receptors located on pancreatic cancer cells and imaging agents due to fluorescence feature of the BSA-HA nanoparticles.</P>

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