Microsatellite instability in patients with chronic B-cell lymphocytic leukaemia.

The purpose of our study was to evaluate the microsatellite instability (MSI) at selected loci with known involvement in the oncogenesis of chronic B-cell lymphocytic leukaemia (B-CLL). DNA from B cells (tumour cells) and from T cells (normal controls) of 27 samples of 26 patients with previously untreated B-CLL was extracted. Microsatellite instability in six microsatellite markers was tested using GeneScan Analysis Software. The rate of replication errors positive phenotype (RER+) was determined (MSI in more than 30% of examined loci). RER+ was found in four out of 27 patients (14.8%). A larger proportion of patients with stage C B-CLL exhibited RER+ than those with stage A or B (P < 0.05). A higher prevalence of RER+ was demonstrated in a subgroup of patients with additional malignancies (three out of eight patients) in comparison with patients with B-CLL alone (1/19) (P = 0.031). In conclusion, our study demonstrated that MSI might have a more prominent role in pathogenesis of B-CLL than reported to date. This may result from a selection of microsatellite markers adjacent to chromosomal loci, which are involved in B-cell malignancies, and using GeneScan Analysis Software, which is most modern and precise method of microsatellite analysis.

Induction of apoptosis by chlorodeoxyadenosine and dexamethasone in B-chronic lymphocytic leukemia.

Apoptosis, programmed cell death, occurs in a variety of cellular systems and in response to many different stimuli. In the present study we examined the ability of dexamethasone (Dex) and chlorodeoxyadenosine (2-CdA) to induce apoptosis in lymphocytes of patients with B-chronic lymphocytic leukemia (B-CLL). Lymphocytes of 29 untreated patients and 9 healthy controls were isolated and incubated for 24 hours in the presence or absence of either Dex (2 microM) (n = 15) or 2-CdA (3 microM) (n = 14). Following incubation the cells were harvested and their DNA extracted and analysed for internucleosomal DNA cleavage by UV illumination after electrophoresis on agarose slab gel containing ethidium bromide. In the Dex group, 10 patients showed dexamethasone independent spontaneous apoptosis appearing 24 hours after the start of incubation. These were the only instances of dexamethasone-enhanced apoptosis. Five patients showed no spontaneous or dexamethasone induced apopto sis. Of the 2-CdA group, 5 showed spontaneous apoptosis enhanced by 2-CdA. No spontaneous apoptosis was observed in the cells from 9 other patients, however, 2-CdA induced apoptosis in 8 cases in this group. This study shows that monitoring of apoptosis in CLL may provide important information regarding susceptibility of the cells to drug induced apoptosis.

Apoptosis as an indicator for steroid sensitivity of lymphocytes in B-chronic lymphocytic-leukemia.

Apoptosis, programmed cell death, occurs in a variety of cellular systems and in response to many different stimuli. One group of apoptosis inducers are glucocorticosteroids which are also found in the battery of cytotoxic drugs used to treat CLL. In the present study we have examined the potency of the glucocorticosteroid-dexamethasone to induce apoptosis in lymphocytes of patients with B-CLL. Lymphocytes of 15 nontreated patients and 5 controls were isolated and incubated for 24 h in the presence or absence of dexamethasone (2 mu M) Following incubation the cells were harvested and their DNA extracted. The extracted DNA samples were analysed for internucleosomal DNA cleavage by UV illumination after electrophoresis on agarose slab gel containing ethidium bromide. Five patients showed neither spontaneous nor dexamethasone induced apoptosis. Whereas, 10 patients, showed a dexamethasone-non-dependent spontaneous apoptosis which appeared 24 h after the start of incubation. The cells of these patients were the only ones to respond to dexamethasone showing an enhanced apoptosis effect. This study shows that apoptosis monitoring in CLL may provide important information regarding susceptibility of the cells to steroid induced apoptosis.