Title Morfološke, imunohistokemijske i kromosomske promjene u stanicama multiplog mijeloma
Title (english) Morphological, immunohistochemical and chromosomal alterations in myeloma cells
Author Irena Seili-Bekafigo
Mentor Toni Valković (mentor)
Committee member Antica Duletić-Načinović (član povjerenstva)
Committee member Ika Kardum-Skelin (član povjerenstva) MBZ: 144581
Committee member Toni Valković (član povjerenstva)
Granter University of Rijeka Faculty of Medicine (Department of General Pathology and Pathological Anatomy) Rijeka
Defense date and country 2014, Croatia
Scientific / art field, discipline and subdiscipline BIOMEDICINE AND HEALTHCARE Clinical Medical Sciences Pathology
Universal decimal classification (UDC ) 616 - Pathology. Clinical medicine
Thesaurus (MESH - Medical Subject Headings )
Multiple Myeloma
Bone Marrow
pathology
Plasma Cells
Abstract Cilj istraživanja: multipli mijelom (MM) je novotvorina plazma stanica, heterogena u
prezentaciji, biološkom ponašanju i odgovoru na liječenje. Zato je neophodno što ranije u
dijagnostičkom procesu svrstati bolesnike u prognostičke grupe i liječenje planirati
individualno. Citološka punkcija koštane srži (KS) jedan je od prvih koraka u dijagnostici
MM. Cilj ovog istraživanja bio je usporediti citomorfologiju i morfometrijske parametre
plazma stanica (PS) sa citogenetskim, molekularnim i kliničkim prognostičkim parametrima
bolesnika sa MM. Hipoteza ovog istraživanja je da bi se na temelju nekih od analiziranih
morfoloških parametara mijelomskih stanica moglo u najranijoj fazi dijagnostičkog postupka
izdvojiti potencijalno prognostički nepovoljnije bolesnike.
Ispitanici i metode: na uzorku od 90 novo dijagnosticiranih bolesnika sa MM usporedili smo
morfološke karakteristike (struktura kromatina, paranuklearno prosvjetljenje, binukleacija i
multinukleacija, istaknuti nukleoli, nepravilne jezgre, centralno smještene jezgre,
vakuolizacija citoplazme, «flaming» morfologija, «pupanje» citoplazme, Russellova tjelešca i
načupani rub citoplazme) i morfometrijske karakteristike plazma stanica (PS) u punktatu KS
(najveći promjer jezgre i citoplazme i omjer tih parametara) sa poznatim prognostičkim
čimbenicima: kliničkim stadijem bolesti po Durie-Salmonu (DS) i po International Staging
System-u (ISS), izražajem NF-κB i Ki-67 odredjenih imunohistokemijski te citogenetskim
promjenama: del(13q14), del(13q34), del17, t(14;16) i t(4;14) odredjenih metodom
fluorescentne in situ hibridizacije (FISH). Također je analizirana medjusobna povezanost
citogenetike, izražaja NF-κB i Ki-67 te kliničkih stadija.
Rezultati: sa kliničkim stadijima statistički značajnu povezanost su pokazali: broj PS u KS,
zrela morfologija PS, načupani rub citoplazme, «flaming» morfologija, «pupanje» citoplazme,
nepravilne jezgre, paranuklearni «halo». Načupani rub citoplazme i manji omjer promjera
VI
jezgre i citoplazme značajno su povezani sa del(13q14), a veći udio PS sa više jezgara sa
del(13q34). Russelova tjelešca češće su nađena uz t(4;14). Izražaj NF-κB povezan je sa
načupanim rubom citoplazme, a Ki-67 sa krupnijim jezgrama, anizocitozom i
anizonukleozom, nepravilnim jezgrama, većim brojem jezgara u PS, i paranuklearnim
«haloom». Izražaj NF-κB bio je značajno veći u nižim kliničkim stadijima po DS, a izražaj
Ki-67 nije povezan sa kliničkim stadijima. Klinički stadij bolesti po ISS pokazao je samo
trend prema povezanosti sa učestalošću del(13q14) i sa ukupnom učestalošću kromosomskih
abnormalnosti, dok klinički stadij po DS nije pokazao značajnu povezanost niti sa jednom
citogenetskom promjenom.
Zaključak: pojedine morfološke i morfometrijske karakteristike PS koreliraju s poznatim
prognostičkim čimbenicima u MM. Zbog toga, morfološka analiza punktata koštane srži
prilikom postavljanja dijagnoze može doprinijeti svrstavanju bolesnika u prognostičke
skupine i dati preliminarnu orjentaciju o potrebi agresivnijeg pristupa liječenju. Kao osobito
značajan morfološki parametar izdvojio se načupani rub PS.
Abstract (english) Objectives: Multiple myeloma (MM) is a plasma cell neoplasm with a very heterogeneous
presentation, biologic behaviour and response to therapy. It is very important to define a risk
group for every patient as soon as possible, in order to choose a personalized therapy
approach. Bone marrow (BM) aspiration is one of the first steps in the diagnostic process for
MM. The aim of this study is to compare cytomorphologic and morphometric features of
plasma cells (PC) with cytogenetic, molecular and clinical prognostic factors in myeloma
patients. The hypothesis is that, maybe, it could be possible to select potentially high risk
myeloma patients at the very beginning of the diagnostic workup, based on some
morphological features of PCs.
Patients and methods: In a cohort of 90 myeloma patients, PCs from BM aspirates at the
moment of diagnosis, were analyzed for series of morphologic and morphometric features:
chromatine structure, paranuclear halo, binucleation/multinucleation, nucleoli, irregular nuclei, centrally placed nuclei, cytoplasmatic vacuolization, flaming PCs, cytoplasmatic
budding, Russell bodies, ragged cytoplasm, percentage of PCs in BM, largest nuclear and
cytoplasmatic diameters and the ratio of the largest nuclear and cytoplasmatic diameters of
PCs. These features were then compared to known prognostic parameters: clinical stage of the
disease according to Durie-Salmon (DS) and the International Staging System (ISS),
expression of NF-κB and Ki-67 in PCs, and some cytogenetic alterations in PCs assessed by
fluorescence in situ hybridization (FISH). The relationship among cytogenetic changes, NF-
κB and Ki-67 expression and clinical stages was analyzed as well.
Results: Clinical stages of MM showed correlation with a percentage of PCs, mature
morphology, ragged cytoplasm, flaming PCs, cytoplasmatic budding, irregular nuclei and
paranuclear halo. Ragged cytoplasm and lesser nuclear/cytoplasmatic diameter ratio were
connected to del(13q14), while a higher percentage of binuclear/multinuclear PCs was
connected to del(13q34) and del(13q14). Russell bodies were more frequently found in
association with t(4;14). NF-κB expression was associated with ragged cytoplasm, and Ki-67
with larger nuclei, anisocytosis and anisonucleosis, irregular nuclei, bi and multinucleation of
PCs and paranuclear halo. NF-κB expression was higher in lower clinical stages according to
DS, and Ki-67 expression did not show any correlation to clinical stages. ISS clinical stages
only showed a tendency towards significant correlation with del(13q14) and with the overall
incidence of chromosomal abnormalities whereas DS clinical stages did not show significant
correlation with any of the chromosomal abnormalities.
Conclusion: Some of the analyzed morphologic and morphometric PC features showed
correlation with known prognostic factors in MM. Thus, morphologic analysis of BM
aspirates at the moment of diagnosis can help in recognising high risk myeloma patients and
provide preliminary information regarding the selection of therapeutic approach. Ragged
cytoplasm of PCs turned out to be quite a significant morphologic feature.
Keywords
multipli mijelom
koštana srž
plazma stanice
citomorfologija
morfometrija
interfazni FISH
NF-κB
Ki-67
klinički stadiji.
Keywords (english)
multiple myeloma
bone marrow
plasma cells
cytomorphology
morphometry
interphase FISH
NF-κB
Ki-67
clinical stages.
Language croatian
URN:NBN urn:nbn:hr:184:535982
Study programme Title: Biomedicine Postgraduate (doctoral) study programme Study programme type: university Study level: postgraduate Academic / professional title: doktor/doktorica znanosti, područje biomedicine i zdravstvo (doktor/doktorica znanosti, područje biomedicine i zdravstvo)
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Created on 2016-03-02 09:42:03