耐三尖杉酯碱白血病细胞株HL-60/H的建立及其耐药性和抗凋亡作用的检测

The Establishment of Multidrug resistance Leukemia Cell Line HL 60/H and Its Biologic Features

为了建立性能稳定的白血病耐药细胞株用于肿瘤细胞耐药性和促凋亡作用的研究， 通过逐渐增高三尖杉酯碱（Ｈ） 的剂量， 用极限稀释法克隆筛选出耐三尖杉酯碱（Ｈ） 的白血病细胞株ＨＬ－６０／Ｈ， 并用ＦＣＭ、透射电镜、ＤＮＡＬａｄｄｅｒ法、ＳＡＢＣ法对其进行耐药性和抗凋亡作用的检测。结果显示， ＨＬ－６０／Ｈ是亲代细胞ＨＬ－６０ 耐受Ｈ 的８９．２ 倍，ＨＬ－６０／Ｈ 在无药培养基中传代１６代， ＩＣ５０ 没有明显的降低。ＨＬ－６０／Ｈ细胞同时对多种药物产生耐受。Ｐ－１７０ 和ＢＣＬ－２在ＨＬ－６０／Ｈ 细胞的表达明显增多。药物不易在细胞内聚集，Ｈ不易诱导其发生凋亡。分析结果认为ＨＬ－６０／Ｈ是性能稳定的耐药细胞株，对耐药机制的研究和寻找逆转耐药的方法提供有利条件；多药耐药蛋白Ｐ－１７０ 介导的药物不易在细胞内聚集可能是ＨＬ－６０／Ｈ细胞产生耐药的重要机制； ＢＣＬ－２ 的高表达使ＨＬ－６０／Ｈ

In order to study drug resistance and drug induced apoptosis, a multidrug resistance leukemia cell line HL 60/H resistant to harringtonine (H) was cloned and screened by using limit diluting method gradually increasing dosage of H Biologic features of HL 60/H were measured by means of FCM, transmission electron microscope, DNA fragmentation assay and SABC The results showed that HL 60/H cells resistant to H were 89 2 fold of HL 60 cells to H After 16 times of subculture in the medium without containing H,IC 50 of H for HL 60/H was not markedly decreased HL 60/H cells were cross resistant to many other kinds of anti cancer drugs The expression of P 170 and BCL 2 was obviously increased in HL 60/H cells The intracellular DNR accumulation in HL 60/H was much less than that in HL 60 It is not easy for H to induce apoptosis of HL 60/H cells It was concluded that HL 60/H is a stable multidrug resistance cell subline The important mechanism for HL 60/H cells resistant to drugs might be that drugs do not easily accumulate in cells because of the overexpression of P 170 HL 60/H cells are resistant to apoptosis induced by drugs because of the overexpression of BCL 2