白细胞介素2和α干扰素逆转白血病细胞多药耐药的研究

目的：探讨细胞因子对人白血病细胞系Ｋ５６２／Ｓ及其多药耐药细胞系Ｋ５６２／Ａ０２的影响。方法：以ＭＴＴ法测定小剂量细胞因子作用后柔红霉素（ＤＮＲ）的毒性变化；用荧光法测定细胞内药物浓度；用免疫组化法检测ｐ－膜糖蛋白（ｐ－ｇｐ）变化；用ＲＴ－ＰＣＲ法检测多药耐药基因（ｍｄｒ－１）ｍＲＮＡ。结果：发现ＤＮＲ对Ｋ５６２／Ａ０２及Ｋ５６２／Ｓ的半数抑制率（ＩＣ５０）分别为４５．０８μｇ／ｍｌ及０．６０７μｇ／ｍｌ。α干扰素（ＩＦＮ－α）和白细胞介素２（ＩＬ－２）作用２４小时可增加ＤＮＲ对Ｋ５６２／Ａ０２的细胞毒作用，与未加药组相比ＩＣ５０下降为１６．３９及１１．９６μｇ／ｍｌ，但不影响Ｋ５６２／Ｓ细胞（ＩＣ５０无明显改变）。且ＩＦＮ－α、ＩＬ－２可提高细胞内ＤＮＲ浓度，从未加药组的２１５１ｎｇ／ｍｇ蛋白到２５７０及２５０３ｎｇ／ｍｇ蛋白。但ｐ－ｇｐ及ｍｄｒ－１ｍＲＮＡ无明显改变。结论：ＩＦＮ－α、ＩＬ－２可增加ＤＮＲ对Ｋ５６２／Ａ０２细胞的毒性作用，增加Ｋ５６２／Ａ０２细胞内的药物浓度，但不是通过下调ｍｄｒ－１ｍＲＮＡ机制而起逆转作用。

Effects of cytokines on multidrug-resistance in K562/A02 cells]

OBJECTIVE: To explore the effects of cytokines on human leukemic cell line K562/S and its multidrug-resistant counterpart K562/A02. METHODS: The toxicities of cytokines and the IC50 (the concentration causing 50% inhibition of cell growth) of DNR were assayed by MTT method; intracellular drug concentration was measured by fluorometry; p-glycoprotein (p-gp) expression was detected by APAAP and mdr-1 mRNA was assayed by RT-PCR. RESULTS: The IC50 of DNR for K562/A02 and K562/S cells were 45.08 microg/ml and 0.607 microg/ml, respectively. Pretreating K562/A02 cells with rhu IFN (500 U/ml) or rhu IL-2 (250 U/ml) for 24 hours partially restored the sensitivity of K562/A02 cells to DNR (IC50 were 16.39 and 11.96 microg/ml, respectively) but had not effect on K562/S cells, and it elevated the intracellular DNR accumulation in K562/A02 from 2151 ng/mg x protein to 2570 and 2503ng/mg x protein, respectively. p-gp and mdr-1 mRNA were not down regulated. By contrast, rhu G-CSF and rhu GM-CSF had no effect on either K562/A02 or K562/S. CONCLUSION: rhu IFN or rhu IL-2 could partially restore the sensitivity of K562/A02 to DNR and elevate the intracellular DNR accumulation via a mechanism independent of p-gp or mdr-1 mRNA down-regulation.