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• W2597802287 abstract "Introduction: Metallothioneins (MT) belong to the group ofintracellular, low-molecular cysteine-rich proteins withmolecular weight from 6 to10 kDa. Due to their high affinity toheavy metal ions (Zn, Cd, As, etc.), their main and crucialfunction is homeostasis maintenance and detoxification of heavymetals. The role of MT in tumour tissue remains still unclear,but MT can be considered as new promising tumour marker.Alpha-methylacyl-CoA racemase (AMACR) is a -oxidation ofbranchedâperoxisomal and mitochondrial enzyme involved in thefatty acids and was recently reported that AMACR has very highsensitivity and specificity to prostate adenocarcinoma.Therefore is very important to find new approaches in AMACRdetection. Aim: The main aim of this paper is to studyexpression of MT and AMACR in tumor prostate cell lines and inpatients serum on protein and mRNA level. Moreover, we comparethe differences in MT expression between cells influnced withzinc ions. Material and methods: Biological samples. Prostaticcell lines derived from prostate adenocarcinoma (LNCaP-FGC,PC-3, and 22RVL) and prostatic cell line derived from normalepithelium (PNT1A – human immortalized prostatic cell line)were used. Protein detection. SDS-PAGE electrophoresis andWestern-blot analysis with subsequent immunodetection wereused. Immunohistochemical detection: Immunohistochemicaldetection of AMACR was performed by R.T.U Vectastain universalABC kit (Vector Laboratories, Burlingame, CA, USA). Results: Weinvestigated influence of zinc ions on MT expression inprostatic cell lines derived from prostate carcinoma and MTlevel in patients serum samples. For the MT determination weused SDS-PAGE electrophoresis and western blot analysis withsubsequent immunodetection. Moreover we used modernelectrochemical methods (Brdicka reaction) for detection of MT.We demonstrated up-regulation of prostatic specific antigenexpression in prostate tumour cells (LNCaP, PC-3, 22RVL) andcontrariwise down-regulation of MT expression. This fact can beexplain by decreased concentration of zinc ions in prostatictumor cells and therefor lower concentration of MT in tumorcell lines in comparing with non-tumor cells. Besidesimmunodetection we measured MT by adsorptive transfer strippingtechnique coupled with differential pulsed voltammetry Brdickareaction and we obtained similar results as in immunodetection.In case of AMACR, results show relatively big diferencesbetween tumor and non-tumor cell lines in AMACR content in mRNAand protein level as well. In case of tumor cells theexpression of AMACR was up-regulated in comparison with nontumour cells. Then we cultivated cells on glass slides forimmunodetection of AMACR in situ and we obtained similarresults. Conclusions: This study provides important newinformation about expresion of MT and AMACR in prostate tumorcell lines. Evidently MT expression is significantly downregulated in 22RVL, and PC-3 cells while AMACR was upregulated. We observed significant (a=0.05) difference in MTcontent between cell lines treated/nontreated with zinc ions byusing immunohistochemical methods and electrochemical methodsas well." @default.
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• W2597802287 date "2011-04-29" @default.
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• W2597802287 title "Determination of metallothioneins and alpha-methylacyl-CoAracemase in tumor prostate diseases" @default.
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