外源性白介素12逆转绒毛膜癌细胞JAR/MTX耐药的作用

Study of IL-12's reverse effect on methotrexate resistant human choriocarcinoma cell in vitro

目的探讨外源性白介素12(IL-12)对绒癌耐药细胞株JAR/MTX的逆转耐药效应。方法将体外培养的绒癌耐药细胞株JAR/MTX分为四组:(1)对照组;(2)化疗组;(3)细胞因子组;(4)化疗+细胞因子组。加药后培养12、24 h,用MTT法检测药物的细胞毒性,用RT-PCR及Western Blot法检测MDR1mRNA和P-gp的表达情况,用流式细胞术检测细胞内底物萤光值强度的变化。结果 (1)外源性IL-12对JAR/MTX的生长抑制作用明显强于甲氨蝶呤(MTX),两药联用后细胞对MTX的敏感性增强。(2)四组细胞MDR1mRNA及P-gp的表达量比较没有明显差别(P>0.05)。(3)加入外源性IL-12的细胞株,胞内底物荧光值强度显著增加(P<0.05)。结论外源性IL-12可通过下调细胞内P-gp的活性来逆转绒癌耐药细胞株JAR/MTX对MTX的耐药效应。

Objective To discuss interleukin-12 （IL-12） ＇ s effectiveness of reversion on methotrexate （MTX） resistant human choriocarcinoma cell line JAR/MTX. Methods JAR/MTX cells were divided into 4 group ： （ 1 ） Control group ; （2） chemothera- peutics group; （3） cytokine group; （4） chemotherapeutics and cytokine group. Each group was observed at 12- and 24-hour. MTT assay was used to detect the cytotoxic effect. The gene expressions of MDR1 and P-gp were detected by Western blot and RT-PCR. The Change in fluorescence intensity was observed by fluorescent test. Results （ 1 ） Interleukin-12＇ s IC50 was lower and relatively sensitive. （2）There were no significant differences in the expressions of MDR1 mRNA and P-gp among all the groups （P 〉 0.05）. （3） There was noticeable difference in fluorescence intensity between the cytokine （IL-12） group and the control one （P 〈 0.05）. Conclusion IL-12 can reverse drug-resistance of JAR/MTX to methotrexate by down-regulating the activity of intracellular P-gp.