title: SLCO1B3 screening in colorectal cancer patients using High-Resolution Melting Analysis method and immunohistochemistry.
abstract: Personalized medicine has made some major advances in colorectal cancer, but new biomarkers still remain a hot issue as an emerging tool with potential prognostic and therapeutic potential. We investigated for SLCO1B3 gene alterations and protein expression in colorectal cancer, using the novel high-resolution melting analysis technique and immunohistochemistry. Formalin-fixed paraffin-embedded tumor samples from 30 colorectal cancer patients were used. The screening for gene alterations was done by high-resolution melting analysis for all exons of SLCO1B3 gene. Organic anion-transporting polypeptide 1B3 protein expression was assessed by immunohistochemistry using the monoclonal mouse MDQ antibody. High level of polymorphism was observed in the SLCO1B3 gene. We identified three previously reported polymorphisms in exons 7, 12, and 14, 699G>A, 1557A>G, and 1833G>A, respectively. In the exon 5, one variant seems to correspond to an as yet unknown SLCO family member. The immunohistochemical study revealed that organic anion-transporting polypeptide 1B3 was expressed in 27/30 samples. Of great interest, the three samples, which were immunohistochemically negative, all appeared to accommodate mutations which lead to either early stop codons or other conformations of the tertiary protein structures affecting the antibody-epitope binding. The results of this study are of much interest as high-resolution melting analysis proved to be a reliable and rapid genotyping/scanning method for mutation detection of SLCO1B3 gene.
authors.affiliation: 1 Cancer Biobank Center, University of Ioannina, Ioannina, Greece.
authors.forename: Lampri
authors.initials: L
authors.lastname: Evangeli
authors.affiliation: 1 Cancer Biobank Center, University of Ioannina, Ioannina, Greece.
authors.forename: Sainis
authors.initials: S
authors.lastname: Ioannis
authors.affiliation: 1 Cancer Biobank Center, University of Ioannina, Ioannina, Greece.|2 Hypoxia and Angiogenesis Group, Molecular Oncology Laboratories, Weatherall Institute of Molecular Medicine, University of Oxford, John, Radcliffe Hospital.
authors.forename: Kounnis
authors.initials: K
authors.lastname: Valentinos
authors.affiliation: 4 Department of Forensic Pathology, University of Ioannina, Ioannina, Greece.
authors.forename: Mitselou
authors.initials: M
authors.lastname: Antigony
authors.affiliation: 3 Department of Pathology, "G. Hatzikosta" General Hospital, Ioannina, Greece.
authors.forename: Ioachim
authors.initials: I
authors.lastname: Elli
authors.affiliation: 1 Cancer Biobank Center, University of Ioannina, Ioannina, Greece.|5 Department of Hematology, Medical School, University of Ioannina, Ioannina, Greece.
authors.forename: Hatzimichael
authors.initials: H
authors.lastname: Eleftheria
authors.affiliation: 6 Departments of Embryology, Histology and Anatomy, University of Ioannina, Ioannina, Greece.
authors.forename: Galani
authors.initials: G
authors.lastname: Vasiliki
authors.affiliation: 1 Cancer Biobank Center, University of Ioannina, Ioannina, Greece.|5 Department of Hematology, Medical School, University of Ioannina, Ioannina, Greece.
authors.forename: Briasoulis
authors.initials: B
authors.lastname: Evangelos
mesh_terms: D000368:Aged; D014408:Biomarkers, Tumor / Q000096:biosynthesis / Q000235:genetics; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000378:metabolism*; D004273:DNA, Neoplasm / Q000032:analysis / Q000235:genetics; D005260:Female; D005557:Formaldehyde; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008875:Middle Aged; D029362:Organic Anion Transporters, Sodium-Independent / Q000096:biosynthesis* / Q000235:genetics*; D016612:Paraffin Embedding; D017422:Sequence Analysis, DNA / Q000379:methods; D000074542:Solute Carrier Organic Anion Transporter Family Member 1B3; D016707:Tissue Fixation
chemical_list: D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm; D029362:Organic Anion Transporters, Sodium-Independent; C421209:SLCO1B3 protein, human; D000074542:Solute Carrier Organic Anion Transporter Family Member 1B3; D005557:Formaldehyde
keywords: Colon cancer; HRMA; SLCO1B3; mutation detection