Tumor necrosis factor-alpha and expression of the multidrug resistance-associated genes LRP and MRP.
Tumor necrosis factor-alpha and expression of the multidrug resistance-associated genes LRP and MRP.
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DOI：
10.1093/jnci/89.11.807
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发表时间：
1997-06
期刊：
影响因子：
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通讯作者：
U. Stein;W. Walther;C. Laurencot;G. Scheffer;R. Scheper;R. Shoemaker
中科院分区：
文献类型：
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作者：
U. Stein;W. Walther;C. Laurencot;G. Scheffer;R. Scheper;R. Shoemaker
表达p -糖蛋白、多药耐药相关蛋白（MRP）或肺耐药蛋白（LRP）的癌细胞已经显示出对多种化疗药物的耐药性。最近，我们报道了表达所有三种蛋白的人结肠癌细胞在暴露于肿瘤坏死因子- α（一种由免疫系统细胞产生的激素样蛋白）后表现出p -糖蛋白基因表达降低和多药耐药性丧失。在本研究中，我们检测了肿瘤坏死因子- α对同一结肠癌细胞中MRP和LRP基因表达的影响。方法将HCT15和HCT116结肠癌细胞与肿瘤坏死因子α (tumor necrosis factor- α)在100 U/mL浓度下孵育2、12、24、48和72小时；另外，用含有人肿瘤坏死因子- α互补DNA的表达载体转染细胞进行研究。采用逆转录和聚合酶链反应方法评价肿瘤坏死因子- α对MRP和LRP信使RNA表达的影响；利用特异性单克隆抗体和流式细胞术检测对MRP和LRP蛋白表达的影响。流式细胞术数据采用双侧、非参数Mann-Whitney秩和检验进行分析。结果外源性肿瘤坏死因子- α治疗降低了两种细胞类型中LRP信使RNA的水平，并呈明显的时间依赖性；在HCT15细胞中，48小时后几乎没有检测到LRP信使RNA。相比之下，HCT116细胞的MRP信使RNA水平升高，而HCT15细胞的MRP信使RNA水平不变。外源性肿瘤坏死因子α治疗诱导两种细胞类型中LRP和MRP蛋白表达的变化，与信使RNA的变化相似。在转染的细胞中，无论细胞类型如何，内源性肿瘤坏死因子- α的产生降低了LRP基因（信使RNA和蛋白质）的表达，增加了MRP基因（信使RNA和蛋白质）的表达。结论在人结肠癌细胞中，肿瘤坏死因子α以相反的方式影响MRP和LRP基因的表达。LRP基因表达的发现与我们早期在这些细胞中p -糖蛋白表达的发现相似。在制定克服肿瘤细胞多药耐药的策略时，应考虑一种药物抑制一种或多种耐药机制并增强其他机制的可能性。
BACKGROUND AND PURPOSE Cancer cells that express P-glycoprotein, multidrug resistance-associated protein (MRP), or lung resistance protein (LRP) have demonstrated resistance to a wide variety of chemotherapeutic drugs. Recently, we reported that human colon carcinoma cells that express all three proteins exhibit reduced P-glycoprotein gene expression and a loss of multidrug resistance after exposure to tumor necrosis factor-alpha, a hormone-like protein produced by cells of the immune system. In this study, we examined the effects of tumor necrosis factor-alpha on MRP and LRP gene expression in the same colon carcinoma cells. METHODS HCT15 and HCT116 colon carcinoma cells were incubated with tumor necrosis factor-alpha at 100 U/mL for 2, 12, 24, 48, or 72 hours; alternatively, cells transfected with an expression vector containing a human tumor necrosis factor-alpha complementary DNA were studied. The effects of tumor necrosis factor-alpha on MRP and LRP messenger RNA expression were evaluated by means of reverse transcription and the polymerase chain reaction; effects on MRP and LRP protein expression were examined by use of specific monoclonal antibodies and flow cytometry. The flow cytometry data were analyzed by use of the two-sided, nonparametric Mann-Whitney rank sum test. RESULTS Treatment with exogenous tumor necrosis factor-alpha reduced the level of LRP messenger RNA in both cell types in an apparently time-dependent fashion; in HCT15 cells, almost no LRP messenger RNA was detected after 48 hours of treatment. In contrast, the level of MRP messenger RNA was increased in HCT116 cells by such treatment, but the level in HCT15 cells was unchanged. Treatment with exogenous tumor necrosis factor-alpha induced changes in LRP and MRP protein expression in the two cell types that paralleled the changes found for messenger RNA. In transfected cells, the endogenous production of tumor necrosis factor-alpha reduced LRP gene expression (both messenger RNA and protein) and increased MRP gene expression (both messenger RNA and protein), regardless of cell type. CONCLUSION In human colon carcinoma cells, tumor necrosis factor-alpha influences MRP and LRP gene expression in opposite ways. The findings for LRP gene expression parallel our earlier findings for P-glycoprotein expression in these cells. IMPLICATION In developing strategies for overcoming multidrug resistance in tumor cells, the possibility that an agent can suppress one or more mechanisms of drug resistance and enhance others should be considered.