scholarly journals Proteomics-based analysis of differentially expressed proteins in the CXCR1-knockdown gastric carcinoma MKN45 cell line and its parental cell

2013 ◽  
Vol 45 (10) ◽  
pp. 857-866 ◽  
Author(s):  
Wanming Hu ◽  
Junpu Wang ◽  
Gengqiu Luo ◽  
Baihua Luo ◽  
Chang Wu ◽  
...  
Keyword(s):  
Gastric Carcinoma ◽  
Cell Line ◽  
Parental Cell ◽  
Differentially Expressed ◽  
Differentially Expressed Proteins ◽  
Mkn45 Cell

scholarly journals iTRAQ Quantitative Analysis of Multidrug Resistance Mechanisms in Human Gastric Cancer Cells

2010 ◽  
Vol 2010 ◽  
pp. 1-11 ◽  
Author(s):  
Huai-Dong Hu ◽  
Feng Ye ◽  
Da-Zhi Zhang ◽  
Peng Hu ◽  
Hong Ren ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Multidrug Resistance ◽  
Cell Line ◽  
Molecular Mechanisms ◽  
Absolute Quantification ◽  
Differentially Expressed ◽  
Human Gastric Cancer ◽  
Differentially Expressed Proteins ◽  
Gastric Cancer Cells ◽  
Proteomic Approach

Multidrug resistance (MDR) is a major obstacle towards a successful treatment of gastric cancer. However, the mechanisms of MDR are intricate and have not been fully understood. To elucidate the molecular mechanisms of MDR in gastric cancer, we employed the proteomic approach of isobaric tags for relative and absolute quantification (iTRAQ), followed by LC-MS/MS, using the vincristine-resistant SGC7901/VCR cell line and its parental SGC7901 cell line as a model. In total, 820 unique proteins were identified and 91 proteins showed to be differentially expressed in SGC7901/VCR compared with SGC7901. Several differentially expressed proteins were further validated by western blot analysis. Furthermore, the association of MVP, one of the highly expressed proteins in SGC7901/VCR, with MDR was verified. Our study is the first application of iTRAQ technology for MDR mechanisms analysis in gastric cancer, and many of the differentially expressed proteins identified have not been linked to MDR in gastric cancer before, which showed the value of this technology in identifying differentially expressed proteins in cancer.

Global Proteomic Profiling of Endemic Versus Sporadic Epstein-Barr Virus (EBV) Positive Burkitt Lymphoma (BL).

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 4779-4779
Author(s):  
Nader El-Mallawany ◽  
Janet Ayello ◽  
Nancy Day ◽  
Carmella van de Ven ◽  
Kevin P Conlon ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
B Cell ◽  
Cell Line ◽  
Cell Lines ◽  
High Energy ◽  
Differentially Expressed ◽  
Differentially Expressed Proteins ◽  
Apoptosis Pathway ◽  
Cellular Processes ◽  
Ebv Infection

Abstract Abstract 4779 Background EBV infection of normal B-cells is commonly associated with the pathogenesis of BL (Brady et al, Clin Path, 2007). Endemic BL (eBL) is characteristically positive (100%) for EBV, contrasting with sporadic BL (sBL), where approximately 30% of cases are positive for EBV. eBL vs. sBL have significantly different breakpoint regions within c-myc (Shiramizu/Magrath et al, Blood, 1991). Overexpression of c-myc is the sine quae non of BL. C-myc interactions with other genes/proteins is multilayered and complex (Basso/Della-Favera, Nat Gen, 2005). Apoptotic pathway disruption is propelled by EBV and is critically important in c-myc deregulation and subsequent lymphomagenesis that occurs in EBV+ eBL vs. sBL (Ruf et al, J Vir, 2000). Global analysis of proteins expressed in EBV+ eBL vs. sBL may provide insights into biologic, pathogenetic, and molecular differences between the two subtypes of lymphoma, and potentially identify targets for the development of therapeutic agents. Objectives To compare the proteomic expression profile and signal transduction pathways of EBV+ eBL vs. sBL. Methods Whole cell lysates obtained from the EBV+ eBL cell line Raji and the EBV+ sBL cell line NC37 were digested and labeled with iTRAQ” labeling reagents, following manufacturer's protocol. The peptides were resolved by 2D-LC technique (off-line Strong cation exchange followed by on-line reverse-phase liquid chromatography). Data-dependant High energy C-trap Dissociation MS/MS spectra were acquired using an Orbitrap XL Tandem Mass Spectrometer (ThermoFisher). The MS/MS data was searched using X!Tandem/TPP software suite against human IPI database (v3.50) appended with decoy (reverse) sequences. iTRAQ” ratios of proteins (ProteinProphet probability of >0.9) were normalized and differentially expressed proteins were determined through Mixture Modeling. Protein interactions were further analyzed using the GoMiner and Ingenuity pathway analysis tools. Results Over 400 proteins were identified as being differentially expressed by a ≥ 1.25 fold change between the two cell lines. We identified differentially expressed proteins in both cell lines that are involved in a wide array of cellular processes as exhibited in Figure 1. Cellular processes uniquely involved by proteins over-expressed in eBL included immune response, hematopoiesis, cell proliferation, heat shock, and B-cell activation, while those uniquely identified in sBL included cell division, response to virus, and NF-kB cascade proteins. Specific cell-regulatory pathways implicated by the differential protein profile expressions (with associated proteins in parentheses) included the p53 apoptosis pathway (PCNA, MSH6, C1QBP, MAP4, and BAX), the caspase network of apoptosis (HCLS1, ACIN1, and AIFM1), the tumor suppressor protein RB network (MCM7, PA2G4, and API5), general apoptotic pathways (HSP90 and PDCD4), B-cell differentiation and proliferation pathways (TPD52 and IKBKG), and the ubiquitin-proteasome pathway (UBE2J1, UBE2C, and UBE2S). Seven of these proteins are c-myc target genes. Ingenuity protein network analysis revealed nine proteins identified in the experiment with interactions connected through the p53, caspase, and tumor necrosis factor apoptosis pathways. Conclusion Proteomic profile analysis of EBV+ eBL and sBL revealed over and under-expression of multiple proteins that may be implicated in the multi-factorial nature of disease pathogenesis. This is the first MS-based direct proteomic comparison of eBL and sBL. Our results suggest that there are potentially different mechanisms driving cell proliferation and resistance to apoptosis in eBL versus sBL and that EBV infection may be involved in the processes that drive lymphomagenesis. Ultimately, identification of proteins unique to the distinct disease subtypes will serve to establish tumor markers that may enable development of new diagnostic, prognostic, and therapeutic strategies. Disclosures: No relevant conflicts of interest to declare.

Quantitative Proteomic Identification of Novel Diagnostic Biomarkers and Therapeutic Targets for Classical Hodgkin Lymphoma.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1462-1462
Author(s):  
Mihaela D. Chiselite ◽  
Damian Fermin ◽  
Venkatesha Basrur ◽  
Kevin P. Conlon ◽  
Charles Seiler ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Cell Line ◽  
Cell Lines ◽  
Total Cell ◽  
Negative Regulator ◽  
Differentially Expressed ◽  
Tandem Mass ◽  
Differentially Expressed Proteins ◽  
Classical Hodgkin Lymphoma ◽  
False Discovery

Abstract There is significant morphologic, immunophenotypic and molecular overlap between some subtypes of diffuse large B-cell lymphoma and classical Hodgkin lymphoma (cHL). In addition, recent studies reveal similar gene expression profiles in PMBCL and cHL suggesting a common cell of origin. We utilized a differential isotopic strategy to determine the global proteomic differences between cell lines derived from cHL (L428), DLBCL (SUDHL-9) and PMBCL (Karpas 1106P). Protein was collected from cell lysates and subjected to labeling by isobaric tags (iTRAQ) for relative quantification and analyzed by reverse-phase liquid chromatography and electrospray ionization tandem mass spectrometry. 200 mg of total cell lysates from each cell line was used and liquid chromatography and tandem mass spectrometry (LC-MS/MS) analyses were performed in duplicate. The proteins were identified using X!Tandem. After normalization, quantitative data were subjected to false discovery rate (FDR) calculation to identify differentially expressed proteins through Mixture Modeling. Significantly differentially expressed proteins were scored at a false discovery rate (FDR) cut-off of ≤ 0.13. This approach yielded 66 proteins with differential expression patterns that were discriminative for the 3 different cell lines. Several proteins that have been previously reported to be differentially expressed between cHL and PMBCL were identified including Fascin, Galectin-1, Galectin-3, STAT1 and SWAP70. In addition, several previously unreported proteins involved in numerous cellular functions were differentially expressed (cell adhesion, signaling, immunity). We have validated a subset of these by Western blot (WB) analysis and immunohistochemistry. For example, the π class of glutathione S-transferases (GSTP), was over-expressed six-fold in K1106P cells vs. L428 cells, and virtually absent in the SUDHL-9 cell line, as confirmed by WB. This enzyme class has been shown to be overexpressed in many human cancers and to be involved in therapy refractoriness. More recently, GSTP has been implicated as a negative regulator of cellular death/apoptosis through the MAP kinase pathway. These factors, and its unusual absence in the SUDHL-9 cell line made GSTP a rational candidate for further functional analysis. Briefly, SUDHL-9 cells and the cHL cell lines L428 and KMH2 were exposed to increasing concentrations (20–80 μM) of the GSTP inhibitor and thiol modifier molecule ethacrynic acid (EA), and subjected to viability assays (WST-1) and to WB for apoptotic markers. Both cHL cell lines were equally resistant to EA concentrations up to 60μM by WST-1 assays, while SUDHL-9 cells were exquisitely sensitive, undergoing total cell necrosis at the lowest (20 μM) concentration. WB analysis for markers of apoptosis showed a higher level of apoptosis in SUDHL-9 cells compared to the HL-derived KMH2 and L428 cells, as evidenced by poly (ADP-ribose) polymerase (PARP) cleavage into its 85 kDa apoptosis-related fragment. L428 cells, expressing 40% the GSTP levels of KMH2 cells by WB, were more susceptible to apoptosis at 60μM EA than KMH2 cells, as evidenced by decrease in S-phase Kinase-associated Protein 2 (p45SKP2), with concomitant increase in p27Kip1, a negative regulator of G1-S progression. These results demonstrate an inverse relationship between GSTP levels and susceptibility to EA-induced cell death in lymphoma. This study demonstrates the utility of large-scale mass spectrometry-based proteomics for the discovery of proteins that may serve as potential diagnostic marker panels for the distinction of PMBCL, DLBCL and cHL, as well as candidate therapeutic targets.

Mass Spectrometry-based Label-free Quantitative Proteomic Analysis of CCl4-induced Acute Liver Injury in Mice Intervened by Total Glycosides from Ligustri Lucidi Fructus

2020 ◽  
Vol 17 ◽  
Author(s):  
Qian Lu ◽  
Hai-Zhu Xing ◽  
Nian-Yun Yang
Keyword(s):  
Insulin Resistance ◽  
Liver Injury ◽  
Protein Expression ◽  
Signaling Pathway ◽  
Proteomic Analysis ◽  
Acute Liver Injury ◽  
Liver Cells ◽  
Differentially Expressed ◽  
Liver Protein ◽  
Differentially Expressed Proteins

Background: CCl4 acute liver injury (ALI) is a classical model for experimental research. However, there are few reports involved in the fundamental research of CCl4-induced ALI Ligustri Lucidi Fructus (LLF) are and its prescription have been used to treat hepatitis illness clinically. LLF and its active ingredients displayed anti-hepatitis effects, but the mechanism of function has not been fully clarified Objective: To investigate the proteomic analysis of CCl4-induced ALI, and examine the effects of active total glycosides (TG) from LLF on ALI of mice4, including histopathological survey and proteomic changes of liver tissues, and delineate the possible underlying mechanism. Methods: CCl4 was used to produce ALI mice model. The model mice were intragastrically administrated with TG and the liver his-topathological changes of mice were examined. At the end of test, mice liver samples were collected, after protein denaturation, re-duction, desalination and enzymatic hydrolysis, identification was carried out by nano LC-ESI-OrbiTrap MS/MS technology. The data was processed by Maxquant software. The differentially-expressed proteins were screened and identified, and their biological information was also analyzed based on GO and KEGG analysis. Key protein expression was validated by Western blot analysis Results: A total of 705 differentially-expressed proteins were identified during the normal, model and administration group. 9 signifi-cant differential proteins were focused based on analysis. Liver protein expression changes of CCl4-induced ALI mice were mainly involved in several important signal channels, namely FoxO signaling pathway, autophagy-animal, insulin signaling pathway. TG has anti-liver damnification effect in ALI mice, the mechanism of which is related to FoxO1 and autophagy pathways Conclusion: CCl4 inhibited expression of insulin-Like growth factor 1 (Igf1) and 3-phosphoinositide-dependent protein kinase 1 (Pdpk1) in liver cells and induced insulin resistance, thus interfered with mitochondrial autophagy and regeneration of liver cells and the metabolism of glucose and lipid, and caused hepatic necrosis in mice. TG resisted liver injury in mice. TG adjusted the expression level of key proteins Igf1 and Pdpk1 after liver injury and improved insulin resistance, thus promoted autophagy and resisted the liver damage

scholarly journals Differentially expressed proteins in platelets derived from patients with hypertension

2021 ◽  
Author(s):  
Yobana Armenta-Medina ◽  
Ivette Martínez-Vieyra ◽  
Oscar Medina-Contreras ◽  
Claudia G. Benitez-Cardoza ◽  
Albertana Jiménez-Pineda ◽  
...  
Keyword(s):  
Differentially Expressed ◽  
Differentially Expressed Proteins
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