ABSTRACT
As one of the hallmarks of cancer, metabolic reprogramming leads to cancer progression, and targeting glycolytic enzymes could be useful strategies for cancer therapy. By screening a small molecule library consisting of 1320 FDA-approved drugs, we found that penfluridol, an antipsychotic drug used to treat schizophrenia, could inhibit glycolysis and induce apoptosis in esophageal squamous cell carcinoma (ESCC). Gene profiling and Ingenuity Pathway Analysis suggested the important role of AMPK in action mechanism of penfluridol. By using drug affinity responsive target stability (DARTS) technology and proteomics, we identified phosphofructokinase, liver type (PFKL), a key enzyme in glycolysis, as a direct target of penfluridol. Penfluridol could not exhibit its anticancer property in PFKL-deficient cancer cells, illustrating that PFKL is essential for the bioactivity of penfluridol. High PFKL expression is correlated with advanced stages and poor survival of ESCC patients, and silencing of PFKL significantly suppressed tumor growth. Mechanistically, direct binding of penfluridol and PFKL inhibits glucose consumption, lactate and ATP production, leads to nuclear translocation of FOXO3a and subsequent transcriptional activation of BIM in an AMPK-dependent manner. Taken together, PFKL is a potential prognostic biomarker and therapeutic target in ESCC, and penfluridol may be a new therapeutic option for management of this lethal disease.
ABSTRACT
<p><b>OBJECTIVE</b>To identify ubiquitinated proteins from complex human multiple myeloma (MM) U266 cells, a malignant disorder of differentiated human B cells.</p><p><b>METHODS</b>Employing a globally proteomic strategy combining of immunoprecipitation, LC-MS/MS and SCX-LC-MS analysis to identified ubiquitination sites, which were identified by detecting signature peptides containing a GG-tag (114.1 Da) and an LRGG-tag (383.2 Da).</p><p><b>RESULTS</b>In total, 52 ubiquitinated proteins containing 73 ubiquitination sites of which 14 and 59 sites contained LRGG-tag and GG-tag were identified, respectively.</p><p><b>CONCLUSION</b>Classification analysis by of the proteins identified in the study based on the PANTHER showed that they were associated with multiple functional groups. This suggested the involvement of many endogenous proteins in the ubiquitination in MM.</p>
Subject(s)
Humans , Cell Line, Tumor , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Physiology , Multiple Myeloma , Metabolism , Neoplasm Proteins , Genetics , Metabolism , Proteomics , Methods , UbiquitinationABSTRACT
Objective:To identify differentially expressed proteins related with malignant transformation of esophageal squamous cell carcinoma (ESCC) using proteomic analysis. Methods:Two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization timE-of flight mass spectrometry (MALDI-TOF-MS) in combination with protein database searching were used to determine and identify differentially expressed proteins in esophageal cancer cell lines (EC1, EC18, and EC109) and immortal cell line (NECA-E6E7-hTERT). Western blotting and immunocytochemistry were used to verify the differential expression of annexin 2 in esophageal cancer cell lines and immortal cell line (NECA-E6E7-hTERT). Real-time fluorogentic quantitative PCR(RFQ-PCR) was performed to analyze the expression level of annexin A2 mRNA.Results: A total of 15 differentially expressed proteins were identified with more than 5 folds difference. Among them three proteins were down-regulated and 12 proteins were up-regulated. Western blotting and immunocytochemical analysis verified the down-regulation of annexin A2 protein in ESCC cell lines. However, differential expression pattern of annexin A2 mRNA was not consistant with its protein expression in ESCC cell lines and immortal cell line (NECA-E6E7-hTERT). Conclusion:The findings provide important clues for identifying the candidate biomarkers for high-risk population screening and early diagnosis of ESCC. Post-translative regulation/modification contributes to the down-regulation of annexin A2 protein.
ABSTRACT
The potential embryotoxicity, developmental toxicity and teratogenic effects of (3 or 8)-(l-methoxyethyl)-8 (or3)-hydroxyethyl-deuteroporphyrin (PsD-044) were investigated in Sprague-Dawley rats with the conventional teratological method in vivo. According to the recommending clinical dosage, PsD-044 was administered intravenously at 20, 10 and 5mg/kg, as compared to the negative control with saline and the positive control with sodium pentachlorophenolate, respectively on the 10th day of the gestation. Eighty-one pregnant rats and 803 fetuses were examined. The results suggest that the maternal toxicity, embryotoxicity and teratogenic effects of PsD-044 were not found, however, the malformation induced by known teratogen was as high as 14.1%.
ABSTRACT
The antimutagenicity of 14 compounds, cysteine (1), cinnamic acid (2), rutin (3), tannic acid (4), germanium dioxide (5),fluoro uracil (6), sodium copper chlorophylline (7),?-sitosterol (8), vitamin C (9), coumarin (10), vitamin E (11), L-glutathine (oxidized form) (12), L-glutathine (reduced form) (13) and organic germanium (14), were investigated using Salmonella typhimurium/microsome assay in TA100.Three modes of action, i.e. inhibitory, antimutagenic and desmutagenic effects, were observed-for their antimutagenicity. Results showed that all of test compounds inhibited mutagenicity induced by MNNG and B(a)P with the exception of germanium dioxide. Percent inhibition of compounds 1, 2, 3, 4, 6, 8, 10 and 12 were greater than 90%. 14 compounds exhibited antimutagenic effects on the mutagenic activity of MNNG, and 12 compounds exhibited antimutagenic effects on B(a)P. Germanium dioxide and organic germanium had no such effect. 14 compounds all exerted desmutagenic effects on B (a)P directly before B(a)P acted on cells. According to the potential and modes of action, cysteine, cinnamic acid, rutin, tannic acid and coumarin were better among 14 compounds. The doseresponse relationships of inhibitory and antimutagenic effects on mutation induced by MNNG and B(a)P were found. Each compound has its sufficient range of dosage. These studies suggest that it is necessary to select effective antimutagens to make mixtures, and their synergistic effects should be investigated.