Enhanced survival in perineural invasion of pancreatic cancer: an in vitro approach
Enhanced survival in perineural invasion of pancreatic cancer: an in vitro approach
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DOI：
10.1016/j.humpath.2006.08.002
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发表时间：
2007-02-01
期刊：
影响因子：
3.3
通讯作者：
Ayala, Gustavo E.
中科院分区：
文献类型：
--
作者：
Dai, Hong;Li, Rile;Ayala, Gustavo E.
胰腺癌（PanCa）以神经周围浸润（PNI）、早期淋巴结转移和肝转移为特征，预后差。PNI是局部复发的重要原因之一。关于PNI在PanCa中的作用机制知之甚少。我们提出了一个新的模型系统，可能揭示了神秘的PNI在PanCa。在本研究中，将小鼠背根神经节（DRGs）和人PanCa细胞系（MIA PaCa-2）在Matrigel基质（BD Biosciences，圣何塞，加利福尼亚州）中共培养以构建该PNI模型。单独的MIA PaCa-2细胞系（对照1）或单独的DRG（对照2）与作为对照的Matrigel基质一起培养。在倒置显微镜下观察神经突生长、细胞集落生长、神经突-集落接触和逆行延伸，然后用Optimas成像系统（Optimas Corp.，博瑟尔，马萨诸塞州）。在第14天，收获实验样品和对照2样品，进行总RNA分离并固定在石蜡包埋的块中。石蜡切片进行Ki-67免疫组化染色和TUNEL检测。使用互补DNA微阵列进行基因谱分析。通过定量逆转录聚合酶链反应验证过表达的靶基因。结果表明，共培养24小时后，神经突与MIA PaCa集落之间存在相互作用。在胰腺癌细胞的存在下刺激神经突生长，其显示比对照2多2倍的面积。72小时后，与DRG共培养的MIA PaCa集落显示出比对照1多58%的集落面积。DRG/MIA PaCa细胞的Ki-67指数（平均值，5.02%）显著高于对照1（平均值，1.18%）（P < .05）;相反，DRG/MIA PaCa细胞的凋亡指数（平均值，0.45%）显著低于对照1（平均值，1.85%）（P < .001）。与对照组相比，DRG/MIA PaCa中的促生存基因MALT 1和TRAF增加2倍。我们证明了神经上皮相互作用是神经和PanCa细胞生长的互利过程。这是可能的，癌基因和生长因子可能协同作用，促进增殖和/或抑制凋亡，生存策略的发展PNI在PanCa的。(c)2007年爱思唯尔公司All rights reserved.
Pancreatic cancer (PanCa) is characterized by perineural invasion (PNI), early lymph node and liver metastasis, and poor prognosis. PNI is one of the important causes of local recurrence. Little is known about the mechanism of PNI in PanCa. We presented a novel model system that may shed light on the mystery of PNI in PanCa. In this study, mouse dorsal root ganglia (DRGs) and human PanCa cell line (MIA PaCa-2) were cocultured in Matrigel matrix (BD Biosciences, San Jose, CA) to build this PNI model. MIA PaCa-2 cell line alone (control 1) or DRG alone (control 2) was cultured with Matrigel matrix as controls. Neurite outgrowth, cell colony growth, neurite-colony contact, and retrograde extension were observed under inverted microscopy and then were photographed and quantitated with the Optimas imaging system (Optimas Corp., Bothell, MA). At day 14, both the experimental and control 2 samples were harvested and subjected to total RNA isolation and fixed in paraffin-embedded blocks. Slides cut from paraffin blocks were studied with Ki-67 immunostaining and TUNEL assay. Gene profiling was performed using complementary DNA microarray. Overexpressed target genes were verified by quantitative reverse transcriptase polymerase chain reaction. The results showed that reciprocity was observed between neurites and MIA PaCa colonies with 24 hours of coculture. Neurite outgrowth was stimulated in the presence of pancreatic carcinoma cells, which showed 2-fold more area than did control 2. After 72 hours, MIA PaCa colonies cocultured with DRG exhibited 58% more colony area than did control 1. The Ki-67 index of the DRG/MIA PaCa cells (mean, 5.02%) was significantly higher than that in control 1 (mean, 1.18%) (P < .05); in contrast, the apoptotic index in the DRG/MIA PaCa cells was significantly lower (mean, 0.45%) than that in the control 1 (mean, 1.85%) (P < .001). Prosurvival genes MALT1 and TRAF were increased 2-fold in DRG/MIA PaCa compared with controls. We demonstrated that neural-epithelial interaction is a mutually beneficial process for the growth of nerves and PanCa cells. It is possible that oncogenes and growth factors might act synergistically in promoting proliferation and/or inhibiting apoptosis, a survival strategy crucial to the development of PNI in PanCa. (c) 2007 Elsevier Inc. All rights reserved.