Department of Medical Microbiology, Immunology, & Cell Biology, Simmons Cancer Institute, Southern Illinois University School of Medicine, 913 N. Rutledge Street, Springfield, IL 62794-9626, USA.
Toxicol Appl Pharmacol. 2011 Aug 15;255(1):40-7. doi: 10.1016/j.taap.2011.05.014. Epub 2011 May 26.
Daunorubicin, idarubicin, doxorubicin and epirubicin are anthracyclines widely used for the treatment of lymphoma, leukemia, and breast, lung, and liver cancers, but tumor resistance limits their clinical success. Aldo-keto reductase family 1 B10 (AKR1B10) is an NADPH-dependent enzyme overexpressed in liver and lung carcinomas. This study was aimed to determine the role of AKR1B10 in tumor resistance to anthracyclines. AKR1B10 activity toward anthracyclines was measured using recombinant protein. Cell resistance to anthracycline was determined by ectopic expression of AKR1B10 or inhibition by epalrestat. Results showed that AKR1B10 reduces C13-ketonic group on side chain of daunorubicin and idarubicin to hydroxyl forms. In vitro, AKR1B10 converted daunorubicin to daunorubicinol at V(max) of 837.42±81.39nmol/mg/min, K(m) of 9.317±2.25mM and k(cat)/K(m) of 3.24. AKR1B10 showed better catalytic efficiency toward idarubicin with V(max) at 460.23±28.12nmol/mg/min, K(m) at 0.461±0.09mM and k(cat)/K(m) at 35.94. AKR1B10 was less active toward doxorubicin and epirubicin with a C14-hydroxyl group. In living cells, AKR1B10 efficiently catalyzed reduction of daunorubicin (50nM) and idarubicin (30nM) to corresponding alcohols. Within 24h, approximately 20±2.7% of daunorubicin (1μM) or 23±2.3% of idarubicin (1μM) was converted to daunorubicinol or idarubicinol in AKR1B10 expression cells compared to 7±0.9% and 5±1.5% in vector control. AKR1B10 expression led to cell resistance to daunorubicin and idarubicin, but inhibitor epalrestat showed a synergistic role with these agents. Together our data suggest that AKR1B10 participates in cellular metabolism of daunorubicin and idarubicin, resulting in drug resistance. These data are informative for the clinical use of idarubicin and daunorubicin.
柔红霉素、伊达比星、多柔比星和表柔比星是广泛用于治疗淋巴瘤、白血病以及乳腺癌、肺癌和肝癌的蒽环类药物，但肿瘤耐药性限制了它们的临床疗效。醛酮还原酶家族 1 B10（AKR1B10）是一种 NADPH 依赖性酶，在肝癌和肺癌中过度表达。本研究旨在确定 AKR1B10 在肿瘤对蒽环类药物耐药性中的作用。使用重组蛋白测定 AKR1B10 对蒽环类药物的活性。通过 AKR1B10 的异位表达或通过依帕司他抑制来确定细胞对蒽环类药物的耐药性。结果表明，AKR1B10 将柔红霉素和伊达比星侧链上的 C13-酮基还原为羟基形式。在体外，AKR1B10 将柔红霉素转化为柔红霉素醇，Vmax 为 837.42±81.39nmol/mg/min，Km 为 9.317±2.25mM，kcat/Km 为 3.24。AKR1B10 对伊达比星表现出更好的催化效率，Vmax 为 460.23±28.12nmol/mg/min，Km 为 0.461±0.09mM，kcat/Km 为 35.94。AKR1B10 对具有 C14-羟基的多柔比星和表柔比星的活性较低。在活细胞中，AKR1B10 有效地催化柔红霉素（50nM）和伊达比星（30nM）还原为相应的醇。在 24 小时内，与载体对照相比，AKR1B10 表达细胞中约 20±2.7%的柔红霉素（1μM）或 23±2.3%的伊达比星（1μM）转化为柔红霉素醇或伊达比星醇，而载体对照中分别为 7±0.9%和 5±1.5%。AKR1B10 的表达导致细胞对柔红霉素和伊达比星的耐药性，但抑制剂依帕司他与这些药物表现出协同作用。综上所述，我们的数据表明 AKR1B10 参与柔红霉素和伊达比星的细胞代谢，导致耐药性。这些数据为伊达比星和柔红霉素的临床应用提供了信息。
