细胞黏附分子L1基因沉默对胰腺癌细胞Capan-2神经侵袭能力的影响
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国家自然科学基金(8107205).


Effect of L1 cell adhesion molecule gene silencing on perineural invasion ability of pancreatic cancer Capan-2 cells
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Supported by National Natural Science Foundation of China(8107205).

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    摘要:

    目的 观察沉默细胞黏附分子L1基因(L1CAM)表达后对胰腺癌Capan-2细胞体外神经侵袭能力的影响。方法 应用慢病毒介导的L1CAM-shRNA干扰载体转染胰腺癌Capan-2细胞,以L1CAM-NC转染细胞为对照。分别将L1CAM-shRNA组及L1CAM-NC组细胞与大鼠背根神经节(DRG)、基质胶(matrigel)一起构成共培养神经侵袭模型,倒置显微镜下观察神经突及细胞生长情况并拍照,利用Image pro plus图像分析软件对照片进行分析。结果 共培养模型中,L1CAM-NC对照组细胞向DRG方向不断迁移增殖,包绕神经突起并沿之向DRG爬行,而L1CAM-shRNA组中并未观察到该现象。共培养第3天和第5天时,L1CAM-NC对照组细胞集落面积显著高于L1CAM-shRNA组(P<0.01),但两组的神经突生长差异无统计学意义。结论 干扰L1CAM的表达可能通过抑制细胞集落形成及迁移爬行的方式抑制胰腺癌Capan-2细胞的神经侵袭作用。

    Abstract:

    ObjectiveTo evaluate the effect of L1 cell adhesion molecule (L1CAM) gene expression silencing by short hairpin RNA (shRNA) on perineural invasion of pancreatic cancer Capan-2 cells in vitro. MethodsWe transfected Capan-2 cells with lentivirus-mediated shRNA targeting L1CAM (L1CAM-shRNA) and negative control shRNA (L1CAM-NC), and then the transfected Capan-2 cells were co-cultured with mouse dorsal root ganglia (DRG) in matrigel matrix. The procession of neurite outgrowth and cell colony growth were observed by inverted microscope. Areas of cell colonies and neurites were quantitated using Image pro plus software. ResultsThe cancer cells migrated to DRG and grew around the neurites in the L1CAM-NC/DRG group, which was not observed in the L1CAM-shRNA/DRG group. On day 3 and 5 of co-culture, the area of cell colonies in the L1CAM-shRNA/DRG group was significantly less than that in L1CAM-NC/DRG group (P<0.01); however, there was no difference in neurite outgrowth between the two groups. ConclusionIt is demonstrated that down-regulation of L1CAM can suppress the perineural invasion of Capan-2 cells in vitro by inhibiting cell proliferation and migration.

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  • 收稿日期:2012-05-17
  • 最后修改日期:2012-09-16
  • 录用日期:2012-09-26
  • 在线发布日期: 2012-10-24
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