Ohmine Ken, Kawaguchi Kei, Ohtsuki Sumio, Motoi Fuyuhiko, Ohtsuka Hideo, Kamiie Junichi, Abe Takaaki, Unno Michiaki, Terasaki Tetsuya
Membrane Transport and Drug Targeting Laboratory, Tohoku University Graduate School of Pharmaceutical Sciences , Sendai, Japan.
Division of Hepato-Biliary-Pancreatic Surgery, Tohoku University Graduate School of Medicine , Sendai, Japan.
Mol Pharm. 2015 Sep 8;12(9):3282-91. doi: 10.1021/acs.molpharmaceut.5b00282. Epub 2015 Aug 25.
The purpose of the present study is to identify the determinant(s) of gemcitabine (dFdC)-sensitivity in pancreatic cancer tissues of patients treated with dFdC alone and in pancreatic cancer cell lines exposed to dFdC in vitro. Protein expression levels of 12 enzymes and 13 transporters potentially involved in transport and metabolism of dFdC in pancreatic cancer cell lines and tissues were quantified by means of our LC-MS/MS-based quantitative targeted proteomics technology. Protein expression levels of deoxycytidine kinase (dCK), uridine monophosphate-cytidine monophosphate (UMP-CMP) kinase, cytosolic nucleotidase III (cN-III), and equilibrative nucleoside transporter 1 (ENT1) were significantly correlated with IC50 or 1/IC50 in five cell lines with different sensitivities to dFdC (p < 0.05). Expression levels of the selected proteins in pancreatic cancer tissues of 10 patients with different progression-free survival (PFS) (49-955 days) were quantified, and their relationship with PFS was examined. Only the protein expression level of dCK was significantly correlated with PFS (p < 0.05). Multiple regression analysis was also performed, and combinations of ENT1, UMP-CMP kinase, CTPS1, and dCK were highly correlated with PFS. Our results indicate that the protein expression level of dCK in pancreatic cancer tissue is a good predictor of PFS, and thus dCK may be the best biomarker of dFdC sensitivity in pancreatic cancer patients treated with dFdC, although other proteins would also contribute to dFdC-sensitivity at the cellular level in vivo and in vitro.
本研究的目的是确定单独接受吉西他滨（dFdC）治疗的患者胰腺癌组织以及体外暴露于dFdC的胰腺癌细胞系中吉西他滨（dFdC）敏感性的决定因素。借助我们基于液相色谱-串联质谱的定量靶向蛋白质组学技术，对胰腺癌细胞系和组织中可能参与dFdC转运和代谢的12种酶和13种转运蛋白的蛋白质表达水平进行了定量。在对dFdC具有不同敏感性的5种细胞系中，脱氧胞苷激酶（dCK）、尿苷单磷酸-胞苷单磷酸（UMP-CMP）激酶、胞质核苷酸酶III（cN-III）和平衡核苷转运体1（ENT1）的蛋白质表达水平与IC50或1/IC50显著相关（p<0.05）。对10例无进展生存期（PFS）不同（49 - 955天）的胰腺癌患者组织中所选蛋白质的表达水平进行了定量，并检测了它们与PFS的关系。只有dCK的蛋白质表达水平与PFS显著相关（p<0.05）。还进行了多元回归分析，ENT1、UMP-CMP激酶、CTPS1和dCK的组合与PFS高度相关。我们的结果表明，胰腺癌组织中dCK的蛋白质表达水平是PFS的良好预测指标，因此，dCK可能是接受dFdC治疗的胰腺癌患者中dFdC敏感性的最佳生物标志物，尽管其他蛋白质在体内和体外细胞水平上也对dFdC敏感性有贡献。
