PDZ a LIM domény obsahující protein 2 (PDLIM2), jinak také Mystique či SLIM, je nejnovějším členem tzv. s aktininy asociované LIM proteinové rodiny proteinů, jejíž členové jsou zásadním způsobem zapojeni do regulace celé škály biologických procesů včetně organizace buněčného cytoskeletu, buněčné diferenciace či onkogeneze. Samotný PDLIM2 je cytoskeletálně i nukleárně lokalizovaný protein kódovaný genem Mystique lokalizovaném na chromozomu 8p21, který reguluje stabilitu a aktivitu nejrůznějších transkripčních faktorů, např. NF κB či STAT. Deregulace PDLIM2 souvisí s celou řadu patologických stavů včetně onkogeneze, přičemž hladiny tohoto proteinu mohou korelovat jak se vznikem, tak se supresí nádorů. Hladiny PDLIM2 jsou epigeneticky potlačeny u celé řady nádorů a to v důsledku hypermetylace promotoru genu Mystique, která brání jeho transkripci. Reaktivace PDLIM2 je schopna potlačit tumorigenicitu a navodit smrt nádorových buněk in vitro i in vivo, což indikuje potenciální tumor supresivní roli tohoto proteinu. Na druhou stranu u nádorových buněčných linií odvozených od metastazujících nádorů bylo detekováno výrazné zvýšení mRNA i proteinových hladin PDLIM2 a bylo zjištěno, že PDLIM2 je asociován s progresí nádorů a vznikem metastáz, což spíše nahrává pro onkogenní funkci tohoto proteinu. Hlavním cílem této práce je detailněji shrnout dosavadní poznatky o roli PDLIM2 v průběhu vzniku a vývoje nádorů, dále vyzdvihnout jeho potenciální využití v klinické praxi, především jako možného terapeutického cíle a také nabídnout možná vysvětlení dosud zjištěných rozdílných funkcí tohoto proteinu v onkogenezi.

PDZ and LIM domain containing protein 2 (PDLIM2), also known as Mystique or SLIM, is a member of the actinin – associated LIM family of proteins that play essential roles in cytoskeletone organization, cell differentiation and have been associated with oncogenesis. PDLIM2 is cytoskeletal and nuclear protein encoded by the Mystique gene localized on chromosome 8p21. PDLIM2 regulates stability and activity of several transcription factors, e. g. NF κB or STAT, and its deregulation is associated with several malignancies. PDLIM2 expression has been connected with both tumor suppression and tumorigenesis. PDLIM2 levels are epigenetically suppressed in different cancers due to Mystique promoter hypermetylation that blocks its transcription. PDLIM2 re expression is able to inhibit tumorigenicity and induces tumor cell death both in vitro and in vivo, which suggest potential tumor suppressor role of PDLIM2. On the other hand, PDLIM2 is highly expressed in cancer cell lines derived from metastatic cancer and its expression is associated with tumor progression and metastasis formation, indicating pro oncogenic role of PDLIM2. The aim of this review is to summarize current knowledge on the role of PDLIM2 in tumor formation and development, focusing on its prospective role as therapeutic target and offering potential explanations of its different functions in oncogenesis that were identified so far. Key words: PDLIM2 protein – COP9 signalosome (CSN) – oncogenesis – 5-aza-2’-deoxycytidine – cancer – metastasis This study was supported by Czech Science Foundation (project No. 14-19250S), European Regional Development Fund and the State Budget of the Czech Republic (OP VaVpI – RECAMO CZ.1.05/2.1.00/03.0101), Brno Ph.D. Talent Scholarship programme – Funded by the Brno City Municipality, by the project MEYS – NPS I – LO1413, MH CZ – DRO (MMCI, 00209805) and BBMRI_CZ (LM2010004). The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE “uniform requirements” for biomedical papers. Submitted: 27. 3. 2015 Accepted: 29. 6. 2015

• Klíčová slova
• PDLIM2 protein, 5-aza-2'-deoxycytidin, COP9 signalozom (CSN),
• MeSH
• genové produkty tax metabolismus   MeSH
• karcinogeneze MeSH
• kolorektální nádory metabolismus   MeSH
• lidé MeSH
• lymfom T-buněčný metabolismus   MeSH
• metastázy nádorů MeSH
• mikrofilamentové proteiny * metabolismus   MeSH
• multiproteinové komplexy fyziologie   metabolismus   MeSH
• nádorové proteiny * MeSH
• nádorové supresorové proteiny * MeSH
• nádory prostaty metabolismus   MeSH
• nádory prsu metabolismus   MeSH
• nádory etiologie   genetika   metabolismus   MeSH
• NF-kappa B metabolismus   MeSH
• proteasy fyziologie   metabolismus   MeSH
• proteiny s doménou LIM * metabolismus   MeSH
• transkripční faktory MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

• Publikační typ
• abstrakt z konference MeSH

BACKGROUND/AIM: Pul-down assay is a popular in vitro method for identification of physical interactors of selected proteins. Here, for the first time, we compared three conventional variants of pull-down assay with the streptavidin-modified surface plasmon resonance (SPR) chips for the detection of PDZ and LIM domain protein 2 (PDLIM2) interaction partners. MATERIALS AND METHODS: PDLIM2 protein-protein interactions were analysed by three variants of pull-down assay on streptavidin beads using LC-MS/MS in "Sequential Window Acquisition of all Theoretical fragment ion spectra (SWATH)" mode and compared with LC-SWATH-MS/MS data from SPR chips. RESULTS: The results showed that (i) the use of SPR chip led to comparable data compared to on-column streptavidin beads, (ii) gravity flow and microflow in wash and elution steps provided better results than centrifugation, and (iii) type and concentration of detergent did not significantly affect the interactome data of cancer-associated PDLIM2. CONCLUSION: Our study supports further application of SPR-based affinity purification with SWATH mass spectrometry for reproducible and controlled characterization of cancer-associated interactomes.

• MeSH
• chromatografie kapalinová MeSH
• interakční proteinové domény a motivy genetika   MeSH
• lidé MeSH
• mikrofilamentové proteiny genetika   izolace a purifikace   MeSH
• nádory genetika   patologie   MeSH
• povrchová plasmonová rezonance * MeSH
• proteiny s doménou LIM genetika   izolace a purifikace   MeSH
• streptavidin chemie   MeSH
• tandemová hmotnostní spektrometrie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Current prognostic factors are insufficient for precise risk-discrimination in breast cancer patients with low grade breast tumors, which, in disagreement with theoretical prognosis, occasionally form early lymph node metastasis. To identify markers for this group of patients, we employed iTRAQ-2DLC-MS/MS proteomics to 24 lymph node positive and 24 lymph node negative grade 1 luminal A primary breast tumors. Another group of 48 high-grade tumors (luminal B, triple negative, Her-2 subtypes) was also analyzed to investigate marker specificity for grade 1 luminal A tumors. From the total of 4405 proteins identified (FDR < 5%), the top 65 differentially expressed together with 30 previously identified and control markers were analyzed also at transcript level. Increased levels of carboxypeptidase B1 (CPB1), PDZ and LIM domain protein 2 (PDLIM2), and ring finger protein 25 (RNF25) were associated specifically with lymph node positive grade 1 tumors, whereas stathmin 1 (STMN1) and thymosin beta 10 (TMSB10) associated with aggressive tumor phenotype also in high grade tumors at both protein and transcript level. For CPB1, these differences were also observed by immunohistochemical analysis on tissue microarrays. Up-regulation of putative biomarkers in lymph node positive (versus negative) luminal A tumors was validated by gene expression analysis of an independent published data set (n = 343) for CPB1 (p = 0.00155), PDLIM2 (p = 0.02027) and RELA (p = 0.00015). Moreover, statistically significant connections with patient survival were identified in another public data set (n = 1678). Our findings indicate unique pro-metastatic mechanisms in grade 1 tumors that can include up-regulation of CPB1, activation of NF-κB pathway and changes in cell survival and cytoskeleton. These putative biomarkers have potential to identify the specific minor subpopulation of breast cancer patients with low grade tumors who are at higher than expected risk of recurrence and who would benefit from more intensive follow-up and may require more personalized therapy.

• MeSH
• databáze proteinů MeSH
• imunohistochemie MeSH
• izotopové značení MeSH
• Kaplanův-Meierův odhad MeSH
• karboxypeptidasa B metabolismus   MeSH
• lidé MeSH
• metastázy nádorů MeSH
• nádorové biomarkery genetika   metabolismus   MeSH
• nádory prsu genetika   metabolismus   MeSH
• NF-kappa B metabolismus   MeSH
• proteomika metody   MeSH
• regulace genové exprese u nádorů MeSH
• reprodukovatelnost výsledků MeSH
• stanovení celkové genové exprese metody   MeSH
• stupeň nádoru MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH