Apoptosis of leukemic cells: a case report [Apoptoza leukemijskih stanica - prikaz bolesnika]

Vrbanus, Ljiljana and Sučić, Mirna and Marković-Glamočak, Mirjana and Ries, Sunčica and Gjadrov-Kuveždić, Koraljka and Fabijanić, Iris and Antulov, Jasenka and Petrik, József and Labar, Boris (2010) Apoptosis of leukemic cells: a case report [Apoptoza leukemijskih stanica - prikaz bolesnika]. Collegium Antropologicum, 34 (2). pp. 705-711. ISSN 0350-6134

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Abstract

Transformation of leukemic cells is associated with delay in maturation and in apoptosis, and to altered responsiveness to growth factors. However, some studies have revealed that Fas (CD95/APO1) which mediates apoptotic signal and decrease of anti-apoptotic Bcl-2 are frequently observed in acute myeloid leukemia (AML) M4/M5 leukemic cells. The aim of the study was to compare cytomorphology and cytochemistry of bone marrow (BM) apoptotic leukemic cells to preserved peripheral blood (PB) leukemic cells in our patient, a 76-year-old man with AML-M5b treated at Zagreb University Hospital Center. BM and PB of the AL patient were analyzed after Pappenheim and cytochemical stainings, and leukemic cells were classified according to FAB and WHO classification. Analysis of PB revealed leukocytosis and 80-90% monocytic cells (46% monoblasts, 29% promonocytes and 11% monocytes). Only a few preserved monoblasts and promonocytes were found in BM, together with numerous morphologically altered cells with characteristic chromatin condensation and pyknosis of nucleus, as well as nuclear fragmentation and formation of apoptotic bodies. Thus, cytomorphology of PB leukemic cells pointed to proliferation of immature monocytic cells, and cytomorphology of BM to cell apoptosis. Cytochemistry of PB monocytic cells and BM apoptotic cells confirmed monocytic cell lineage because esterase was strongly positive in almost all BM apoptotic leukemic cells and PB leukemic cells, and esterase was completely inhibited with sodium fluoride. On the basis of these findings, AML-M5b was diagnosed in our patient. There are many possible explanations for our observation of BM leukemic cell apoptosis in a patient with AML-M5. The most reliable one is that apoptosis was induced ex vivo after BM aspiration in course of the air drying of BM specimen before staining. Mass BM leukemic cell apoptosis that was recorded in contrast to numerous preserved leukemic cells in PK could be probably connected to unfavorable ratio of relatively low concentration of cytokines in relation to high leukemic cell number in BM aspirated cytologic specimen.

Abstract in Croatian

Transformacija leukemijskih stanica povezana je s odgodom sazrijevanja, odgodom apoptoze i promijenjenim odgovorom na faktore rasta. Međutim, neke studije su pokazale da se molekula Fas (CD95/APO1), koja provodi signal apoptoze – programirane stanične smrti, te snižene vrijednosti anti-apoptotičke molekule Bcl-2, češće nalaze na leukemijskim stanicama akutnih mijeloičnih lekemija (AML) tipa M4 i M5. Cilj rada bio je usporediti citomorfologiju i citokemiju apoptotičkih leukemijskih stanica koštane srži (KS) i očuvanih leukemijskih stanica periferne krvi (PK) u našeg 76 godina starog bolesnika s M5b koji je liječen u Kliničkom bolničkom centru Zagreb. Koštana srž i periferna krv bolesnika analizirane su nakon standardnog bojenja po Pappenheimu i citokemijskih bojenja i akutna leukemija (AL) je klasificirana prema kriterijima FAB (French-American-British) i SZO (Svjetska zdravstvena organizacija). Analiza PK je pokazala leukocitozu i 80–90% monocitnih stanica (46% monoblasta, 29% promonocita i 11% monocita). U KS su nađene brojne morfološki promijenjene stanice s karakterističnom kondenzacijom kromatina i piknotičkim jezgrama, fragmentacijom jezgri, apoptotička tjelešca, te tek nekoliko očuvanih monoblasta i promonocita. Citomorfologija leukemijskih stanica PK upućivala je na proliferaciju monocitnih stanica, dok je citomorfologija KS upućivala na apoptozu stanica. Citokemija monocitnih stanica PK i apoptotičkih stanica KS potvrdila je monocitnu liniju stanica jer je u gotovo svim leukemijskim apoptotičnim stanicama KS i leukemijskim stanicama PK nespecifična esteraza bila izrazito pozitivna i njena je aktivnost potpuno inhibirana natrijevim fluoridom. Na temelju morfologije i citokemije leukemijskih stanica postavljena je dijagnoza AL-M5b. Brojni poremećaji u leukemijskim stanicama teoretski bi mogli potaknuti apoptozu u stanicama KS u našeg bolesnika s M5b. Do apoptoze je, međutim, najvjerojatnije došlo ex vivo nakon što je učinjena dijagnostička punkcija koštane srži, dok su se citološki uzorci sušili na zraku tijekom 2 sata prije postupka bojenja. Objašnjenje za tako velik broj apoptotičkih leukemijskih stanica u koštanoj srži nasuprot brojnim očuvanim leukemijskim stanicama u PK moglo bi biti povezano s nepovoljnim odnosom relativno niske koncentracije citokina u odnosu na izrazito velik broj leukemijskih stanica u aspiratu koštane srži.

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