摘要
目的:研究三氧化二砷(As2O3)诱导白血病K562/ADM耐药细胞凋亡的分子机制。方法:采用MTT比色法检测K562/ADM耐药细胞增殖活性,细胞形态学和annexinV /PI双染色检测细胞凋亡,RT-PCR检测mdr1、bcl-2和caspase-3基因mRNA的表达水平,流式细胞法(FCM)测定P-糖蛋白(P-glycoprotein,P-gp)和bcl-2蛋白表达及caspase-3活性。结果:As2O3显著抑制K562/ADM耐药细胞的增殖;经As2O3处理后细胞形态上出现典型的凋亡改变,annexinV /PI双染显示凋亡细胞明显增加;mdr1mRNA表达和P-gp合成明显降低,凋亡抑制基因bcl-2mRNA及其蛋白bcl-2表达下调,caspase-3mRNA表达和caspase-3活性显著增强。结论:As2O3诱导K562/ADM耐药细胞凋亡,其主要机制可能为As2O3抑制mdr1和bcl-2基因表达,逆转耐药白血病细胞因bcl-2和P-gp高表达所介导的凋亡抑制。
Objective To explore the molecular mechanisms of AsEO3-induced apoptosis of muhidrug-resistant leukemia K562/ADM cells. Methods The proliferating activity of K562/ADM cells was assessed by MTI＇ assay. Ceil apoptosis was determined by and annexinV/PI double-staining. Expressions of mdrl, bcl-2 and caspase-3 mRNA were examined by reverse transcription polymerase chain reaction （RT-PCR）. Expressions of bcl-2 protein and p-glycoprotein （P-gp） and caspase-3 activity were measured with FCM. Results As203 effectively inhibited the growth of K562/ADM cells. A typical apoptotic change was observed in K562/ADM cells after treatment with As203, and annexinV/PI staining revealed an obvious increase in the apoptosis rate of the cells, the expressions of mdrl and bcl-2 mRNA, P-gp and bcl- 2 protein decreased while caspase-3 mRNA expression and caspse-3 activity markedly increased. Conclusions As203 induce apoptosis of drug-resistant K562/ADM cells. Its major molecular mechanism may be involved in the inhibitory effect of As203 on the expressions of mdrl and bcl-2, and the reversal of inhibition of apoptosis in K562/ADM cells mediated by high expressions of P-gp and bcl-2.
出处
《实用医学杂志》
CAS
2007年第23期3651-3654,共4页
The Journal of Practical Medicine
基金
甘肃省科技攻关计划项目(编号:GS022-A43-137)
