盐酸吉西他滨对K562细胞增殖、凋亡及bcr/abl mRNA表达的影响

Effects of Gemcitabine on Proliferation,Apoptosis and Expression of bcr/abl Fusion Gene mRNA in K562 Cells

目的观察盐酸吉西他滨对K562细胞增殖、凋亡及bcr/abl基因表达的影响,探讨盐酸吉西他滨用于慢性粒细胞白血病急变期治疗的可行性。方法采用水溶性四甲基偶氮唑盐(MTT)比色法检测盐酸吉西他滨对K562细胞增殖的影响,采用凋亡的原位酶标记检测(TUNEL)法检测盐酸吉西他滨对K562细胞凋亡的影响,采用半定量RT-PCR技术检测盐酸吉西他滨作用于K562细胞后bcr/abl融合基因mRNA表达变化。结果0.1 mg/L、1.0 mg/L、10.0 mg/L盐酸吉西他滨分别作用K562细胞12 h、24 h、48 h、72 h后,K562细胞增殖受到抑制,抑制作用随质量浓度及时间增加而增强,但48 h和72 h比较无显著性差异,10.0 mg/L吉西他滨组作用K562细胞48 h抑制率达(29.3±0.008)%。吉西他滨组对K562细胞的抑制作用明显强于同质量浓度同时间的阿糖胞苷组,二者比较有显著性差异(P<0.01)。TUNEL实验结果显示10.0 mg/L吉西他滨作用于K562细胞48 h,凋亡率为15.3%,明显高于阿糖胞苷组3.7%(P<0.05)和空白对照组2.0%(P<0.05)。RT-PCR结果显示10 mg/L吉西他滨作用48 h对K562细胞bcr/abl融合基因mRNA的表达有明显抑制作用,与同剂量阿糖胞苷组比较无显著性差异。结论盐酸吉西他滨对K562细胞有一定的增殖抑制及促凋亡作用,且具有剂量和时间依赖性。10 mg/L吉西他滨能够明显下调慢性粒细胞白血病bcr/abl融合基因mRNA表达,有望联合应用于慢性粒细胞白血病急变期的治疗。

Objective To observe the effects of gemeitabine on proliferation,apoptosis and expression of bcr/abl fusion gene mRNA in K562 cells,and to explore the feasibility that children with chronic myeloid leukemia BP could be treated with gemcitabine at acute transformation phase.Methods The activities and proliferation of K562 cells treated with gemcitabine were examined by MTT assay.The apoptosis of K562 cells induced by gemcitabine was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end lab...