尼莫地平对阿糖胞苷诱导HL60细胞凋亡机制影响的研究_医学论文.docVIP

尼莫地平对阿糖胞苷诱导HL细胞凋亡机制影响的研究_医学论文 尼莫地平对阿糖胞苷诱导HL细胞凋亡机制影响的研究_医学论文 【摘要】 为了研究尼莫地平对阿糖胞苷诱导HL细胞凋亡机制的影响，采用琼脂糖凝胶电泳检测其DNA凋亡带，用细胞免疫组织化学方法检测细胞凋亡相关基因bcl、bax的蛋白表达。结果表明，实验组自培养8小时起琼脂糖凝胶电泳显示典型的DNA梯形凋亡带。Bcl蛋白表达在各实验组随培养时间延长逐渐下降，而Bax蛋白的表达则逐渐增加，Bcl比值逐渐下降，8小时就与对照组出现差异（0.05）。结论：尼莫地平及阿糖胞苷均能促进HL细胞凋亡，诱导其凋亡的机制与下调bcl及上调bax基因的蛋白表达有关。尼莫地平能加强阿糖胞苷促进HL细胞凋亡，其机制与协同下调bcl的表达有关。 【关键词】 尼莫地平 阿糖胞苷 凋亡 HL细胞系 蛋白表达 Bcl Effect of Nimodipine on Mechanisms of HL60 Cell Apoptosis Induced by Cytarabine Abstract The aim was to study the mechanisms of HL60 cell apoptosis induced by nimodipine (NMDP ) and cytarabine (AraC). The DNA fragment was detected by agarose gel electrophoresis. The expressions of bcl2 and bax gene proteins related with apoptosis were investigated by immunohistochemistry. The results showed that HL60 cell apoptosis rate had been increasing in the experimental groups compared with the control group since culturing 8 hours. The expression of Bcl2 protein was lower and the expression of Bax protein was higher in the experimental groups than that in the control group, while ratio of bcl2/bax was lower in the experimental groups than that in the control group. It is concluded that NMDP and AraC induce apoptosis of HL60 cells, and the mechanism of apoptosis induced by them may downregulate the expression of bcl2 gene and upregulate the expression of bax gene. The mechanism of HL60 cell apoptosis induced by AraC and NMDP is probably associated with the downregulation of Bcl2 protein expression. Key word nimodipine cytarabine apoptosis HL60 cell line protein expression Bcl2 Bax 白血病多药耐药性(MDR)的产生是导致化疗失败的一个主要原因［1],凋亡相关基因bcl、bax的表达与白血病耐药之间有着密切关系［2]。bcl表达增高能使白血病细胞抵抗糖皮质激素、VP16、DNR、MIT、Ara、VCR、ADM和ATRA等化疗药物及X射线所诱导的凋亡， bax是调节bcl肿瘤耐药的重要基因之一［3］。研究表明，钙通道拮抗剂尼莫地平(nimodipine, NMDP) 通过加强化疗药物诱导细胞凋亡而达到化疗增强作用［4、5］。NMDP是如何加强化疗药物诱导细胞凋亡的？其作用机制尚不明了。为此，本研究以人类早幼粒细胞白血病细胞株（HL）为研究对象，探讨尼莫地平对阿糖胞苷诱导HL细胞凋亡机制的影响，为临床合理用药提供理论依据。 材料和方法 细胞和主要试剂 HL细胞株引自武汉大学典型培养物保藏中心。NMDP购来自德国拜尔公司，Ara购自上海华联制药厂。Rabbit Anti、Rabbit Anti、SA