ABSTRACT
Objective To explore the role and clinical significance of GSTM 3 ( glutathione S-trans-ferase mu 3) expression in prostate cancer (PCa). Methods We had used the two-dimensional fluores-cence difference gel electrophoresis ( 2D-DIGE) and mass spectral analysis to further verify the microarray data of mRNA expression profiling discovered .GSTM3 mRNA level was detected by Rael-time Quantitative PCR ( RT-QPCR) in 28 pairs of prostate cancer tissue and benign tissue .The relationship of GSTM 3 level with the serum PSA level and the clinical feature of PCa were analyzed . Results In 2D-DIGE study, we found that the expression of GSTM 3 protein in adjacent tissues was significantly higher than that in PCa tis-sues (P0.05) and prostate cancer clinical pathological parameters ( P>0.05). Conclusions GSTM3 expression is down-regulated in PCa tissues, and we may identify PCa by detecting the GSTM 3 expression .
ABSTRACT
Objective To screen the sensitive plasma molecular markers in diabetic nephropathy ( DN) patients with kidney yang deficiency. Methods Two-dimensional fluorescence difference gel electrophoresis (2-D DIGE) was used to analyze the plasma which was sampled from 4 early DN patients with kidney yang deficiency and 4 healthy adult volunteers. Proteins that showed differential expression with an over 1.5-fold change were analyzed by MALDI-TOF/TOF mass spectrometry. Results Two-dimensional electrophoresis (2-DE) maps of plasma proteins in DN patients with kidney yang deficiency and healthy adults were established successfully. We validated 9 differentially expressed proteins, including complements C3 and C4, apolipoprotein E and ubiquitination factor. Conclusion Proteomic analysis by 2-D DIGE can objectively reveal the differences of plasma protein expression in DN patents with kidney yang deficiency and healthy adult volunteers. The obtained 9 proteins have potential to provide plasma molecular markers for the early clinical diagnosis and for the research of traditional Chinese medical patterns of DN.
ABSTRACT
Objective To screen for new methylation association genes in HL-60 to reveal the pathogenesis of leukemia, and provide important theoretical and scientific basis for the prevention and cure of leukemia. Methods Two-dimensional fluorescence difference gel electrophoresis (F-2D-DIGE) was performed to separate the total proteins from acute myelogenous leukemia (AML) cell line HL-60 cells with or without 5-aza-2-deoxycytidine (5-aza-2-dC) treatment. Imaging software Decyder 6.5 and PDQuest were used to detect the differential expression protein spots, and matrix-assisted laser desorption/ionizaion time-of-flight mas spectrometer (MALDI-TOF MS) was adopted to identify the differential expression proteins. Results F-2D-DIGE maps of 5-aza-2-dC-untreated HL-60 and-treated HL-60 cells were established. A total of 53 differential protein spots were detected, and 35 differential proteins were successfully identified. Of the identified proteins, 32 proteins were up-regulated, and 3 proteins were down-regulated in HL-60 cells after 5-aza-2-dC treatment.Conclusion Thirty-five differential proteins may be associated with methylation in HL-60 cell line, which provides the important clues for epigenetic study of leukemia.
ABSTRACT
The mechanism of how stroma plays an important role in tumor carcinogenesis is now a hotspot. To delineate the features of stromal protein between nasopharyngeal carcinoma (NPC) and normal nasopharyngeal mucosa(NNM), laser capture microdissection (LCM) was performed to purify stromal cells from the NPC and NNM, respectively. The protein expressed profiles of the stroma of NPC and NNM were compared using fluorescent two-dimensional difference gel electrophoresis (2D-DIGE) and 34 differential protein spots between tumor stroma and normal stroma were chosen to be identified by mass spectrometry (MS). A total of 20 differential proteins were identified, and three differential proteins (CapG, L-plastin and S100A9) were selectively further validated by Western blotting and immunohistochemical analysis to confirm the results of 2D-DIGE. 2D-DIGE patterns of the stroma of NPC and NNM were established for the first time, the results suggested that differentially expressed proteins in the stroma of NPC and NNM may be useful for understanding the relationship between NPC cells and their surrounding microenvironment. Further studying of these proteins will be helpful to elucidate the mechanisms of NPC carcinogenesis and provide new thoughts on therapy of NPC through stroma.