ABSTRACT
@#【Objective】 To investigate the relationship between the expression of PRDX3 （thioredoxin-dependent peroxide reductase）and the occurrence and development of ccRCC （clear cell renal cell carcinoma）. 【Methods】 The expression of PRDX3 was first verified in 16 cases of ccRCC tissues and adjacent normal tissues. In the present study ， according to the PRDX3 over-expression level，we established the stable PRDX3 overexpression cell lines and knockdown cell lines in 786-O cell lines. We detected the growth rate of tumor cells after overexpression and knockdown of PRDX3. Interaction proteins with PRDX3 were searched by anti-flag pull-down test combined with LC- MS/MS technique. The interaction between PRDX3 and PRDX1（peroxiredoxin 1）was preliminarily explored.【Results】The western blot results showed that PRDX3 were down- regulated in 14 out of 16 ccRCC tissue samples about 1.78 times. Stable PRDX3 overexpression and knockdown cell lines and those control group were successfully established［786O- PRDX3（+）and 786O- PRDX3（-），786O- PRDX3 KN and 786O- PRDX3 NCi］. PRDX3 expression in 786O- PRDX3（+）was 2.1 times higher than 786O- PRDX3（-）at mRNA level and 1.8 times at protein level. PRDX3 expression in 786O- PRDX3 KN was 0.48 times lower than 786O-PRDX3 NCi at mRNA level and 0.51 times at protein level. The cell growth rate of 786O-PRDX3 （+）cell lines was significantly lower than that of 786O-PRDX3（-）. Meanwhile ，there was no significant difference in 786O-PRDX3 KN and NCi cell lines. Pull-down results shows that PRDX3 may interact with PRDX1 through disulfide bond and the binding sites of those two proteins were identified respectively.【Conclusion】PRDX3 was down- regulated expression in renal clear cell carcinoma and the interaction with PRDX1 may be involved in the occurrence and development of tumor. Increasing the expression level of PRDX3 can significantly reduce the growth rate of tumor cells. Based on PRDX3 ，it is possible to develop targeted drugs for treating renal clear cell carcinoma.
ABSTRACT
<p><b>BACKGROUND</b>Multiple myeloma (MM) is a malignant tumor, which takes the second place in malignant blood disease. The clinical symptoms are complicated that make more difficult to diagnose and therapy. Lots of researches focus on the proteins about MM in order to solve those problems. We used proteomic methods to find potential biomarkers in MM patients.</p><p><b>METHODS</b>We applied the peptide ligand library beads (PLLBs) to deplete high abundance proteins in serum for finding potential pathogenic factors and biomarkers of MM. Using 1D-Gel-liquid chromatography-tandem mass spectrometry (LC-MS/MS), we identified 789 and 849 unique serum proteins in MM patients and in healthy controls, respectively.</p><p><b>RESULTS</b>Twenty-two proteins were found differentially expressed between the two groups including serum amyloid A protein, vitamin D-binding protein isoform-1 precursor, plasma kallikrein, and apolipoprotein A-I. Changes of integrin alpha-11 and isoform-1 of multimerin-1 were validated with Western blotting. The linkage of the differentially expressed proteins and the pathogenesis pathways of MM were discussed.</p><p><b>CONCLUSIONS</b>PLLB combined with 1D-gel-LC-MS/MS analysis is an efficient method to identify differentially expressed proteins in serum from patients with MM.</p>
Subject(s)
Humans , Biomarkers , Blood , Biomarkers, Tumor , Blood , Multiple Myeloma , Blood , Peptide Library , Proteomics , Methods , Tandem Mass SpectrometryABSTRACT
<p><b>BACKGROUND</b>The molecular mechanisms underlying the endometriosis are still not completely understood. In order to test the hypothesis that the approaches in phosphoproteomics might contribute to the identification of key biomarkers to assess disease pathogenesis and drug targets, we carried out a phosphoproteomics analysis of human endometrium.</p><p><b>METHODS</b>A large-scale differential phosphoproteome analysis, using peptide enrichment of titanium dioxide purify and sequential elution from immobilized metal affinity chromatography with linear trap quadrupole-tandem mass spectrometry, was performed in endometrium tissues from 8 women with or without endometriosis.</p><p><b>RESULTS</b>The phosphorylation profiling of endometrium from endometriosis patients had been obtained, and found that identified 516 proteins were modified at phosphorylation level during endometriosis. Gene ontology annotation analysis showed that these proteins were enriched in cellular processes of binding and catalytic activity. Further pathway analysis showed that ribosome pathway and focal adhesion pathway were the top two pathways, which might be deregulated during the development of endometriosis.</p><p><b>CONCLUSIONS</b>That large-scale phosphoproteome quantification has been successfully identified in endometrium tissues of women with or without endometriosis will provide new insights to understand the molecular mechanisms of the development of endometriosis.</p>