HL-60细胞及其耐药细胞株HL-60/ADM超弱发光的检测  

The examination of ultraweak luminescence of HL-60 cells and adriamycin-resistant HL-60/ADM cells

作　　者：马世荣[1] 惠延平[1] 徐玉乔[1] 李焰[2] 

机构地区：[1]第四军医大学基础部病理学教研室,陕西西安710032 [2]第四军医大学口腔医学院生物学教研室,陕西西安710032

出　　处：《细胞与分子免疫学杂志》2007年第7期609-611,615,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(60471035)

摘　　要：目的:探讨人急性早幼粒白血病细胞系(HL-60)和其耐药株抗阿霉素人急性早幼粒白血病细胞系(HL-60/ADM)的超弱发光与耐药性的关系。方法:应用MTT比色法、流式细胞术、细胞生长曲线和免疫组化染色法,检测HL-60细胞、HL-60/ADM细胞对阿霉素(ADM)的敏感性、细胞周期变化及P-糖蛋白(P-gp170)的表达。用IFFM-D型流动式化学发光仪检测HL-60细胞及HL-60/ADM细胞的超弱发光强度。结果:HL-60/ADM细胞对ADM的敏感性明显低于亲本HL-60细胞;HL-60/ADM细胞中G0/G1期的细胞为44·80%±1·97%,G2/M期的细胞为9·90%±0·27%,较其亲本细胞增多,S期的细胞为45·30%±1·93%,较其亲本细胞减少(P<0·05);HL-60/ADM细胞倍增时间是HL-60细胞的1·8倍,P-gp170的表达呈阳性,HL-60细胞呈阴性。在1×10-4mol/L鲁米诺及3mL/L双氧水条件下,密度分别为8×107/L、1×108/L、1·26×108/L及2·73×108/L的HL-60/ADM细胞超弱发光强度低于HL-60细胞(P<0·05)。结论:HL-60/ADM细胞对ADM的敏感性降低,细胞生长延缓,P-gp170表达呈阳性等均提示HL-60/ADM细胞出现了耐药性;且随着HL-60/ADM细胞耐药性的产生其超弱发光强度较其亲本细胞HL-60降低,提示细胞超弱发光强度明显降低可作为细胞耐药性产生的一项指标。AIM： To explore the relationship between the ultraweak luminescence and adriamycin resistantance in human leukemic cells （HL-60） and human adriamycin resistant leukemic cells （HL-60/ADM）. METHODS： MTT colorimetry, flow cytometry （ FCM）, growth curve and immunohistochemical staining were used to detect the sensitivity of HL-60 and HL-60/ADM cells to ADM, the change of cell cycle and the expression of P-glycoprotein （P-gp170）. The intensity of the ultraweak luminescence of HL-60 cells and HL-60/ADM cells was examined by the IFFM-D chemiluminescence instrument. RESULTS： The sensitivity of HL- 60/ADM cells to ADM was distinctly lower than that of HL-60 cells. The percentage of HL-60/ADM cells in G0/G1 was 44.80%±1.97% and that in G2/M phase was 9.90% ±0.27%, which was both higher than that of HL-60 cells. The percentage of HL-60/ADM cells in S phase was 45.30% ±.93%, which was lower than that of HL-60 cells. The doubling time of HL-60/ADM cells was 1.8 times as long as that of HL-60 cells. The expression of P-gp170 was positive in HL-60/ ADM cells but negative in HL-60 cells. Treated with 10-4 mol/L luminol and 3 mL/L H2O2 and under four different cell concentration （8 ×10^7/L, 1 ×10^8/L, 1. 26 ×10^8/L, 2.73 ×10^8/L）, the intensity of ultraweak luminescence of HL-60/ ADM cells was distinctly lower than that of HL-60 cells （ P 〈 0.05）. CONCLUSION： The lower sensitivity to ADM, longer doubling time and the expression of P-gp170 showed the drug resistance of HL-60/ADM cells. With the obtaining of drug resistance, ultraweak luminescence intensity was lower in HL-60/ADM cells than that of HL-60 cells. It showed that the decreasing of intensity of ultraweak luminescence could be an index of examining drug-resistantce of tumor cells.

关 键 词：抗阿霉素 超弱发光 耐药性 人急性早幼粒白血病细胞系HL-60 

分 类 号：R735.7[医药卫生—肿瘤]