Profiling SLCO and SLC22 genes in the NCI-60 cancer cell lines to identify drug uptake transporters.
Profiling SLCO and SLC22 genes in the NCI-60 cancer cell lines to identify drug uptake transporters.
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DOI：
10.1158/1535-7163.mct-08-0539
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发表时间：
2008-09
影响因子：
5.7
通讯作者：
Gottesman MM
中科院分区：
文献类型：
--
作者：
Okabe M;Szakács G;Reimers MA;Suzuki T;Hall MD;Abe T;Weinstein JN;Gottesman MM
NCI-60细胞组的分子和药理学分析提供了识别涉及耐药性或敏感性的途径的可能性。其中，外排转运体介导的抗癌药物摄取减少是研究得最好的机制之一。先前的研究也表明，摄取转运体可以通过改变细胞对抗癌药物的摄取来影响细胞毒性。采用实时荧光定量PCR技术，检测了正常组织和NCI-60细胞组中有机阳离子/两性离子转运体（SLC22家族）和有机阴离子转运体（SLCO家族）两个溶质载体（SLC）家族共23个基因的mRNA表达情况。通过将SLCO和SLC22家族成员基因产物的mRNA表达模式与在NCI-60细胞系上测试的1429种抗癌药物和候选药物的生长抑制谱相关联，我们确定了SLC蛋白可能在药物敏感性中起主导作用。为了证实一些SLC -药物对，其中SLC成员被预测会致敏，后续实验使用过表达SLC22A4 （OCTN1，有机阳离子/肉毒碱转运蛋白1）的工程和表征细胞系进行。正如统计相关性预测的那样，SLC22A4的表达导致细胞摄取增加，对米托蒽醌和阿霉素的敏感性提高。我们的研究结果表明，基因表达数据库可用于鉴定对癌细胞敏感的SLCO和SLC22家族成员。
Molecular and pharmacological profiling of the NCI-60 cell panel offers the possibility of identifying pathways involved in drug resistance or sensitivity. Of these, decreased uptake of anticancer drugs mediated by efflux transporters represents one of the best studied mechanisms. Previous studies have also shown that uptake transporters can influence cytotoxicity by altering the cellular uptake of anticancer drugs. Using quantitative real-time PCR, we measured the mRNA expression of two solute carrier (SLC) families, the organic cation/zwitterion transporters (SLC22 family) and the organic anion transporters (SLCO family), totaling 23 genes in normal tissues and the NCI-60 cell panel by quantitative real-time PCR. By correlating the mRNA expression pattern of the SLCO and SLC22 family member gene products with the growth inhibitory profiles of 1,429 anticancer drugs and drug candidate compounds tested on the NCI-60 cell lines, we identified SLC proteins that are likely to play a dominant role in drug sensitivity. To substantiate some of the SLC–drug pairs for which the SLC member was predicted to be sensitizing, follow-up experiments were performed using engineered and characterized cell lines over-expressing SLC22A4 (OCTN1, organic cation/carnitine transporter 1). As predicted by the statistical correlations, expression of SLC22A4 resulted in increased cellular uptake and heightened sensitivity to mitoxantrone and doxorubicin. Our results indicate that the gene expression database can be used to identify SLCO and SLC22 family members that confer sensitivity to cancer cells.