Additional confirmation of successful cell suspension fractionation of metastatic axillary node tissue.

Autor: Ivanović V; Department for Radiobiology and Molecular Genetics, Vinča Institute of Nuclear Sciences, National Institute of the Republic of Serbia, University of Belgrade, Belgrade, Serbia., Tepavčević S; Department for Molecular Biology and Endocrinology, Vinča Institute of Nuclear Sciences, National Institute of the Republic of Serbia, University of Belgrade, Belgrade, Serbia., Dedović-Tanić N; Department for Radiobiology and Molecular Genetics, Vinča Institute of Nuclear Sciences, National Institute of the Republic of Serbia, University of Belgrade, Belgrade, Serbia; State University of Novi Pazar, Department for Natural Sciences and Mathematics, Novi Pazar, Serbia., Milovanović Z; Institute of Oncology and Radiology of Serbia, Belgrade, Serbia., Stojiljković B; Oncology Institute of Vojvodina, Sremska Kamenica, Serbia., Vasiljević T; Oncology Institute of Vojvodina, Sremska Kamenica, Serbia., Mandušić V; Department for Radiobiology and Molecular Genetics, Vinča Institute of Nuclear Sciences, National Institute of the Republic of Serbia, University of Belgrade, Belgrade, Serbia. Electronic address: vvranic@vinca.rs.
Jazyk: angličtina
Zdroj: Pathology, research and practice [Pathol Res Pract] 2024 Aug; Vol. 260, pp. 155439. Date of Electronic Publication: 2024 Jul 01.
DOI: 10.1016/j.prp.2024.155439
Abstrakt: We present herein an extension to our recently developed and published method termed "Fractionation of Nodal Cell Suspension" (FNCS). The method enables efficient subcellular fractionation into nuclear (N) and cytosolic (C) compartments of extremely fibrous and problematic metastatic axillary lymph node (mALN) tissue, using the entire nodule. For the purpose of the present study, a case of invasive lobular breast cancer (BC) patient with pT2N3aMx status and defined primary tumor markers (ERα 8, PR-B 8, and HER2 score 0) was available. Initially, the mALN tissue of this patient was analyzed by immunohistochemistry (IHC), and a positive correlation of nodal ERα, PR-B and HER2 biomarkers to those of the primary tumor was obtained. Subsequently, the mALN was FNCS fractionated into N and C, and Western blot (WB) analysis demonstrated a single band for ERα, PR-B and nuclear loading control (HDAC1) in nuclear, but not in the cytosolic compartments, confirming the efficiency of our fractionation protocol. At the same time, HER2 bands were not observed in either compartment, in accordance with HER2 negativity determined by IHC in both primary tumor and mALN tissue. In conclusion, by confirming the nuclear expression of ERα and PR-B biomarkers in metastatic loci, we demonstrate the purity of the FNCS-generated compartments - the protocol that offers a reliable tool for further analysis of nuclear versus cytosolic content in downstream analysis of novel biomarkers in the whole mALN of BC patients.
Competing Interests: Declaration of Competing Interest All authors declare no competing interests.
(Copyright © 2024 Elsevier GmbH. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: