BCL-2反义硫代磷酸寡脱氧核苷酸对原代急性白血病细胞选择性抑制作用

SELECTIVE INHIBITION OF GROWTH OF ACUTE LEUKEMIC CELLS BY TREATMENT WITH BCL-2 ANTISENSE PHOSPHOROTHIOATEOLIGODEOXYNUCLEOTIDES

为探讨BCL-2反义硫代磷酸寡脱氧核苷酸(AS-PS-ODN,ASPO)对急性原代白血病细胞和正常或良性血液病骨髓细胞的作用是否存在差别。应用台盼蓝拒染试验测定细胞存活力;用造血祖细胞集落培养:粒—单系祖细胞集落(CFU-GM),多向祖细胞集落(CFU-Mix),后红系祖细胞集落(CFU-E)、前红系祖细胞集落(BFU-E)和白血病祖细胞集落(CFU-AML,CFU-ALL)培养检测细胞增殖能力;免疫细胞化学染色检测细胞BCL-2蛋白表达变化。结果发现(1)正常或良性血液病骨髓细胞经10μmol/L ASPO处理一周,同对照组比较:细胞生长数、CFU-GM、CFU-Mix、CFU-E、BFU-E及BCL-2蛋白表达均无显著差别(P<0.05)。(2)急性原代白血病细胞经5μmol/L ASPO处理一周,同对照组比较:细胞生长数显著减低;CFU-AML或CFU-ALL显著降低(P < 0.05),而CFU-GM明显增高(P< 0.05);对照组BCL-2蛋白表达率为77.92％±22.50％,ASPO组于培养的第3天为54.05％±20.20％(P<0.01)和第6天为55.35％±22.74％(P<0.05)均显著低于对照组。因此认为BCL-2ASPO具有选择性地抑制白血病细胞的增殖和BCL-2蛋白的表达的作用。

To observe the different effects between acute leukemic cells and bone marrow cells of normal or benign blood diseases produced by treatment with Bcl-2 antisense phosphorothioate oligodeoxynucleotides(ASPS-ODN, ASPO). The cellular viability was detected by trypan blue exclusion. The ability of proliferation of hematopoietic cells were determined by the culture of colony forming unit-granulocyte/ macrophage(CFU-GM) ,mix-colony forming unit(CFU-Mix) , colony forming unit-erythroid(CFU-E)and burst forming unit-erythroid (BRU-E). The ability of proliferation of leukemic cells were tested by the culture of colony forming unit-acute myeloid leukemia(CFU-AML)and colony forming unit-acute lymphoid leukemia( CFU-ALL). The expressions of Bcl-2 protein was determined by immunocyto-chemistry. It was shown that in normal or benign blood disease, there were no significant differences in the cellular viability, proliferation of hematopoietic cells and the expression of Bcl-2 protein whatever the treatment with ASPO at 10umol/L or without ASPO. In acute leukemia,otherwise; the cellular viability and the forming rate of CFU-AML or CFU-ALL were significant lower in presence of ASPO at 5μmol/L than that without ASPO(P<0.05), but the amount of CFU-GM was significant higher(P<0. 05). The expression rate of Bcl-2 protein of acute leukemic cells treated without ASPO was 77. 92% ±22. 50% . In the third day and the sixth day of treatment with ASPO, the expression rates of Bcl-2 protein of acute leukemic cells were significantly decreased to 54. 05% ± 20. 20% (P<0. 01)and 55.35% ±22.74%(P<0.05)respectively.Our data suggested that ASPO could selectively inhibit the proliferation and the expression of Bcl-2 protein in acute leukemic cells.