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目的:研究8-氯腺苷(8-Choroadenosine,8-Cl-Ado)对人T淋巴母细胞白血病细胞株Molt-4的生长和凋亡的作用,以及对相关基因和CREB结合活性的影响。方法:以不同浓度8-Cl-Ado处理体外培养的Molt-4细胞,采用MTT法测定药物半数抑制浓度(IC_(50)),用光学显微镜、流式细胞仪和琼脂糖电泳分析细胞凋亡,分别用斑点杂交法和免疫组化法观察c-mye和bcl-2的表达情况,用凝胶阻滞电泳检测CREB与DNA结合活性变化。结果:8-Cl-Ado可显著抑制Molt-4细胞的生长,IC_(50)为0.6μmol/L。8-Cl-Ado可诱导Molt-4细胞凋亡,细胞出现典型的凋亡形态学变化,流式细胞分析G_1期细胞数增加,S期细胞数减少并有亚G_1峰,琼脂糖电泳可见DNALadder出现,此外观察到c-myc基因表达瞬时增强和CREB结合活性的增强,但bcl-2基因表达无明显变化。结论:8-Cl-Ado能抑制Molt-4细胞的生长和诱导其凋亡,此生物学效应可能与c-myc瞬时表达增强及CREB结合活性增强有关。该药对molt-4细胞bcl-2基因表达无影响。
AIM: To investigate the effects of 8-chloro-adenosine (8-Cl-Ado) on the growth and apoptosis of human T lymphoblastic leukemia cell line Molt-4 and its effect on the related genes and CREB binding activity. Methods: Molt-4 cells cultured in vitro were treated with different concentrations of 8-Cl-Ado. The half-inhibitory concentration (IC 50) of the drug was determined by MTT assay. Apoptosis was analyzed by light microscopy, flow cytometry and agarose gel electrophoresis The expression of c-mye and bcl-2 were observed by dot blot hybridization and immunohistochemistry respectively. The binding activity of CREB to DNA was detected by gel-block electrophoresis. Results: 8-Cl-Ado significantly inhibited the growth of Molt-4 cells with IC50 of 0.6 μmol / L. The apoptosis of Molt-4 cells was induced by 8-Cl-Ado. The morphological changes of apoptotic cells were observed. Flow cytometry analysis showed that the number of cells in G 1 phase increased, the number of cells in S phase decreased and the subcellular G_1 peak was observed. Appeared, in addition to transiently enhanced expression of c-myc gene and CREB binding activity, but bcl-2 gene expression did not change significantly. Conclusion: 8-Cl-Ado can inhibit the growth and induce the apoptosis of Molt-4 cells. This biological effect may be related to the transient expression of c-myc and the increase of CREB binding activity. The drug had no effect on bcl-2 gene expression in molt-4 cells.