Over-expression of genes and proteins of ubiquitin specific peptidases (USPs) and proteasome subunits (PSs) in breast cancer tissue observed by the methods of RFDD-PCR and proteomics

Abstract

The ubiquitin-proteasome system facilitates the degradation of damaged proteins and regulators of growth and stress response. Alterations in this proteolytic system are associated with a variety of human pathologies. By restriction fragment differential display polymerase chain reaction (RFDD-PCR) and matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF-TOF MS) based on two-dimensional polyacrylamide gel electrophoresis (2-DE), differentially expressed genes and proteins of ubiquitin specific proteases (USPs), proteasome subuinits (PSs) and ubiquitin protein ligase E3A (UBE3A) were analyzed between breast cancer and adjacent normal tissues. Some of them were further verified as over-expression by immunohistochemical stain. Five genes of proteasome subunits (PSs), including PSMB5, PSMD1, PSMD2, PSMD8 and PSMD11, four genes of USPs, including USP9X, USP9Y, USP10 and USP25, and ubiquitin protein ligase E3A (UBE3A) were over-expressed (>3-fold) in breast cancer tissue compared to adjacent normal tissue, and over-expression (>4-fold) of proteins of PSMA1 and SMT3A were observed in breast cancer tissue. PSMD8, PSMD11 and UBE3A were further verified as over-expression by immunohistochemical stain. The action of ubiquitin-proteasome system were obviously enhanced in breast cancer, and selectively intervention in action of ubiquitin-proteasome system may be a useful method of treating human breast cancer.

Validation of UBE2C protein as a prognostic marker in node-positive breast cancer

“…Our results were consistent with those of other studies (Chen and Madura, 2005;Deng et al, 2007). Of the 38 genes included in our 38-gene expression signature, UBE2C was the most highly ranked gene.…”

“…Finally, our IHC study included more patients than did that of Berlingieri (92 instead of 74). In regard to our recent studies and other 'Omics' studies, numerous components of the UPS have been found to be related to breast carcinogenesis and an unfavourable evolution in breast HR = 7.14 (1.64 -31. cancer (Chen and Madura, 2005;Ricolleau et al, 2006;Deng et al, 2007;Campone et al, 2008;Jézéquel et al, 2008). Furthermore, increased activity of the proteasome was directly linked to overexpression of UPS elements (ubiquitin enzymes and proteasomal subunits) (Chen and Madura, 2005).…”

Transcription alterations of members of the ubiquitin–proteasome network in prostate carcinoma

“…In recent studies, proteasome inhibitors display proapoptotic effects. 5,[12][13] The first proteasome inhibitor (bortezomib, Velcade, PS-341) has now been approved for the treatment of advanced multiple myeloma. [14][15] Additionally, trials for other haematological and solid tumours, including PCA have been initiated.…”

High intensity training induces alteration of the ubiquitin-proteasome system gene expression profile and structural changes in the ovaries

“…nos. 600-100, 615-100, 616-100, 617-100, 618-100) provided by Qbiogene, Inc. and MP Biomedicals, Inc. (Carlsbad, CA, USA) as previously described and used in our laboratory (15,16). After RFDD-PCR, 15 µl loading buffer was added, and the samples were heated to 85˚C for 5 min and placed directly on ice.…”