Miyazaki Junichi, Tsuzuki Yoshikazu, Matsuzaki Koji, Hokari Ryota, Okada Yoshikiyo, Kawaguchi Atsushi, Nagao Shigeaki, Itoh Kazuro, Miura Soichiro
Second Department of Internal Medicine, National Defense Medical College, Saitama, Japan.
Int J Cancer. 2005 Nov 10;117(3):499-505. doi: 10.1002/ijc.21202.
Most cases of pancreatic cancer are inoperable when diagnosed. Since immunotherapy and antiangiogenic therapy have been reported to be promising for pancreatic cancer, we examined whether the combination of immunotherapy with dendritic cells (DCs) and the antiangiogenic drug TNP-470 induces tumor regression. Syngeneic mouse pancreatic adenocarcinoma cells were orthotopically inoculated into C57/BL6 mice. DCs with or without tumor lysate (TL) were administered i.p. at 4 and 5 weeks. TNP-470 was injected s.c. into tumor-bearing mice every other day from 4 weeks to 6 weeks. We compared anticancer effects in 6 groups: NT (no treatment), DC/TL- (DCs without TL), DC/TL+ (DCs pulsed with TL), TNP (TNP-470 alone), DC/TL-TNP (DC/TL- plus TNP-470) and DC/TL+TNP (DC/TL+ plus TNP-470). We measured tumor volume, mean vascular density (MVD) and vessel diameter by FITC-dextran using an intravital microscope; degrees of proliferation and apoptosis of cancer cells by PCNA and TUNEL; infiltrating lymphocytes and expression levels of VEGF and MMP-9 by immunohistochemistry and immunoblotting. Tumor volume and MVD were significantly suppressed in the treatment groups with prolonged survival rate, especially in the DC/TL+TNP group. There were no significant differences in apoptosis among the 6 groups except DC/TL+. The number of infiltrating CD4+ cells in the DC/TL+ group was higher than that in the NT group. VEGF expression was significantly suppressed in the treatment groups containing TNP-470, and MMP-9 was also suppressed in the groups containing DC/TL+. Our data suggested that TL-pulsed DCs combined with TNP-470 induced regression of mouse pancreatic cancer, possibly through induction of immune responses and suppression of angiogenesis.
大多数胰腺癌病例在确诊时已无法进行手术。由于免疫疗法和抗血管生成疗法已被报道对胰腺癌有前景，我们研究了免疫疗法与树突状细胞（DCs）和抗血管生成药物TNP - 470联合使用是否能诱导肿瘤消退。将同基因小鼠胰腺腺癌细胞原位接种到C57/BL6小鼠体内。在第4周和第5周腹腔注射含或不含肿瘤裂解物（TL）的DCs。从第4周开始至第6周，每隔一天给荷瘤小鼠皮下注射TNP - 470。我们比较了6组的抗癌效果：NT（未治疗）、DC/TL - （不含TL的DCs）、DC/TL + （用TL脉冲处理的DCs）、TNP（单独使用TNP - 470）、DC/TL - TNP（DC/TL - 加TNP - 470）和DC/TL + TNP（DC/TL + 加TNP - 470）。我们使用活体显微镜通过FITC - 葡聚糖测量肿瘤体积、平均血管密度（MVD）和血管直径；通过PCNA和TUNEL检测癌细胞的增殖和凋亡程度；通过免疫组织化学和免疫印迹检测浸润淋巴细胞以及VEGF和MMP - 9的表达水平。治疗组的肿瘤体积和MVD显著受到抑制，生存率延长，尤其是在DC/TL + TNP组。除DC/TL + 组外，6组之间的凋亡无显著差异。DC/TL + 组中浸润的CD4 + 细胞数量高于NT组。在含TNP - 470的治疗组中VEGF表达显著受到抑制，在含DC/TL + 的组中MMP - 9也受到抑制。我们的数据表明，TL脉冲处理的DCs与TNP - 470联合使用可诱导小鼠胰腺癌消退，可能是通过诱导免疫反应和抑制血管生成实现的。
