Smetana Karel, Pluskalová Michaela, Marinov Yuri, Hrkal Zbynek
Institute of Hematology and Blood Transfusion, Prague, Czech Republic.
Med Sci Monit. 2004 Nov;10(11):BR405-9. Epub 2004 Oct 26.
Since photodynamic treatment (PDT) induces apoptosis in individual HL-60 cells originating from early granulocytic precursors of acute myeloid leukemia, the present study was undertaken to provide information on such treatment of K562 cells, which originate from early granulocytic precursors of chronic myeloid leukemia (blastic phase) and carry the bcr/abl fusion gene with anti-apoptotic properties.
MATERIAL/METHODS: PDT was based on the 5-aminolevulinic acid treatment of K562 cells, followed by blue light irradiation under conditions which in HL-60 cells induce an apoptotic process without previous terminal maturation. Nuclei and nucleoli were visualized by cytochemical procedures to demonstrate DNA, RNA and silver stained proteins of nucleolus organizer regions (AgNORs). TUNEL and propidium iodide assays were used for additional control of the incidence of apoptotic and necrotic cells.
In contrast to HL-60 cells, PDT did not induce apoptosis in K562 cells. However, after PDT some K562 cells exhibited major alterations, expressed by nuclear and cell swelling, reflecting a necrotic process, confirmed by propidium iodide. In addition, PDT produced a reduction of AgNORs, though smaller than that previously described in apoptotic HL-60 cells.
PDT produced only necrotic changes in some K562 cells under conditions which induced in apoptosis in HL-60 cells. Thus the induction of apoptosis, nuclear and nucleolar changes in individual cells does not depend on the inducer--PDT--but on the cell properties.
由于光动力治疗(PDT)可诱导源自急性髓系白血病早期粒细胞前体的单个HL-60细胞发生凋亡,因此开展本研究以提供关于K562细胞这种治疗的信息,K562细胞源自慢性髓系白血病(急变期)的早期粒细胞前体,并携带具有抗凋亡特性的bcr/abl融合基因。
材料/方法:PDT基于对K562细胞进行5-氨基酮戊酸处理,随后在能诱导HL-60细胞发生凋亡过程而无需先前终末成熟的条件下进行蓝光照射。通过细胞化学方法观察细胞核和核仁,以显示核仁组织区(AgNORs)的DNA、RNA和银染蛋白。采用TUNEL和碘化丙啶检测进一步控制凋亡和坏死细胞的发生率。
与HL-60细胞不同,PDT未诱导K562细胞凋亡。然而,PDT后一些K562细胞表现出主要变化,表现为核肿胀和细胞肿胀,这反映了坏死过程,碘化丙啶检测证实了这一点。此外,PDT使AgNORs减少,尽管比先前在凋亡的HL-60细胞中描述的减少幅度小。
在能诱导HL-60细胞凋亡的条件下,PDT仅在一些K562细胞中产生坏死变化。因此,单个细胞中凋亡、核及核仁变化的诱导不取决于诱导剂——PDT——而是取决于细胞特性。