Gemcitabine enhances Wilms’ tumor gene WT1 expression and sensitizes human pancreatic cancer cells with WT1-specific T-cell-mediated antitumor immune response
Gemcitabine enhances Wilms’ tumor gene WT1 expression and sensitizes human pancreatic cancer cells with WT1-specific T-cell-mediated antitumor immune response
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DOI：
10.1007/s00262-011-1033-3
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发表时间：
2011-05
期刊：
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通讯作者：
Akitaka Takahara;S. Koido;Masaki Ito;Eijiro Nagasaki;Yukiko Sagawa;T. Iwamoto;H. Komita;T. Ochi;H. Fujiwara;M. Yasukawa;J. Mineno;H. Shiku;S. Nishida;H. Sugiyama;H. Tajiri;S. Homma
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作者：
Akitaka Takahara;S. Koido;Masaki Ito;Eijiro Nagasaki;Yukiko Sagawa;T. Iwamoto;H. Komita;T. Ochi;H. Fujiwara;M. Yasukawa;J. Mineno;H. Shiku;S. Nishida;H. Sugiyama;H. Tajiri;S. Homma
Wilms肿瘤基因（WT1）在人胰腺癌（PC）中表达，是一种被t细胞介导的抗肿瘤免疫应答识别的独特肿瘤抗原。研究了肝癌标准治疗药物吉西他滨（Gemcitabine， GEM）对肝癌细胞WT1表达的调控及WT1特异性抗肿瘤免疫应答的增敏作用。通过定量PCR、免疫印迹分析和共聚焦显微镜检测WT1的表达。采用质谱法检测HLAⅰ类分子上WT1抗原肽。使用wt1特异性T细胞受体基因转导的人T细胞作为效应T细胞进行细胞毒活性分析。GEM处理人MIAPaCa2 PC细胞后，WT1 mRNA水平升高，这种升高与核因子κ B激活有关。来自gem处理的携带miapaca2的SCID小鼠的肿瘤组织也显示WT1 mRNA的增加。除MIAPaCa2外，部分人PC细胞系在GEM处理后WT1 mRNA水平上调。GEM处理将WT1蛋白从细胞核转移到细胞质中，这可能促进了WT1蛋白的蛋白酶体加工和抗原肽的产生。事实上，与未处理的细胞相比，gem处理的MIAPaCa2细胞中HLA-A*2402限制性WT1抗原肽（CMTWNQMNL）的表达增加。wt1特异性细胞毒性T细胞杀死最佳剂量GEM处理的MIAPaCa2细胞比未处理的MIAPaCa2细胞更有效。GEM增强了WT1在人PC细胞中的表达，并使PC细胞产生WT1特异性t细胞介导的抗肿瘤免疫反应。
Wilms’ tumor gene (WT1), which is expressed in human pancreatic cancer (PC), is a unique tumor antigen recognized by T-cell-mediated antitumor immune response. Gemcitabine (GEM), a standard therapeutic drug for PC, was examined for the regulation of WT1 expression and the sensitizing effect on PC cells with WT1-specific antitumor immune response. Expression of WT1 was examined by quantitative PCR, immunoblot analysis, and confocal microscopy. Antigenic peptide of WT1 presented on HLA class I molecules was detected by mass spectrometry. WT1-specific T-cell receptor gene–transduced human T cells were used as effecter T cells for the analysis of cytotoxic activity. GEM treatment of human MIAPaCa2 PC cells enhanced WT1 mRNA levels, and this increase is associated with nuclear factor kappa B activation. Tumor tissue from GEM-treated MIAPaCa2-bearing SCID mice also showed an increase in WT1 mRNA. Some human PC cell lines other than MIAPaCa2 showed up-regulation of WT1 mRNA levels following GEM treatment. GEM treatment shifted WT1 protein from the nucleus to the cytoplasm, which may promote proteasomal processing of WT1 protein and generation of antigenic peptide. In fact, presentation of HLA-A*2402-restricted antigenic peptide of WT1 (CMTWNQMNL) increased in GEM-treated MIAPaCa2 cells relative to untreated cells. WT1-specific cytotoxic T cells killed MIAPaCa2 cells treated with an optimal dose of GEM more efficiently than untreated MIAPaCa2 cells. GEM enhanced WT1 expression in human PC cells and sensitized PC cells with WT1-specific T-cell-mediated antitumor immune response.