Morphological and cytoskeletal changes of pancreatic cancer cells in three-dimensional spheroidal culture
Morphological and cytoskeletal changes of pancreatic cancer cells in three-dimensional spheroidal culture
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DOI：
10.1007/s00795-010-0497-0
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发表时间：
2010-12-01
影响因子：
1.8
通讯作者：
Naito, Zenya
中科院分区：
文献类型：
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作者：
Matsuda, Yoko;Ishiwata, Toshiyuki;Naito, Zenya
三维（3D）细胞培养有望模拟体内环境。我们使用纳米培养板来确定胰腺导管腺癌（PDAC）细胞系的球状形成能力，并将PDAC细胞的形态和细胞骨架蛋白的表达与二维（2D）培养的细胞进行比较。所有检测的PDAC细胞在二维培养中均呈单层生长。PANC-1和KLM-1在三维培养中形成球体，而PK-45H和MIAPaCa-2则没有形成球体。在3D培养中，附着于平板表面的细胞强烈表达F-actin，形成细胞突起。在PANC-1细胞的纳米培养板网格上检测到F-actin，而在PK-45H细胞中未检测到。三维培养细胞中微管蛋白表达水平高于二维培养。E-cadherin mRNA在PANC-1和KLM-1中的表达水平高于PK-45H和MIAPaCa-2。综上所述，PDAC细胞在三维培养中表现出形态改变、球形形成和细胞骨架蛋白的改变。e -钙粘蛋白可能是参与PDAC细胞球形形成的关键分子之一。三维球体培养系统是一种有用的方法，细胞成像的对比相显微镜和共聚焦显微镜。
Three-dimensional (3D) cell cultures are expected to mimic in vivo environments. We used a NanoCulture plate to determine the spheroid-forming ability of pancreatic ductal adenocarcinoma (PDAC) cell lines and compared the morphology and expression of cytoskeletal proteins of PDAC cells to those in two-dimensional (2D) cultures. All examined PDAC cells grew as monolayers in 2D culture. PANC-1 and KLM-1 formed spheroids in 3D culture, but PK-45H and MIAPaCa-2 did not. Strong expression of F-actin was observed in the cells attached to the surface of the plate, which formed cell projections in 3D culture. F-actin was detected on the grids of the NanoCulture plate in PANC-1 cells but not in PK-45H. The levels of tubulin expression in cells were higher in 3D culture than in 2D culture. The expression level of E-cadherin mRNA in PANC-1 and KLM-1 was higher than that in PK-45H and MIAPaCa-2. In conclusion, PDAC cells showed morphological changes, spheroid formation, and alterations of cytoskeletal proteins in 3D culture. E-cadherin might be one of the key molecules involved in spheroid formation of PDAC cells. The 3D spheroidal culture system was a useful method for cell imaging with contrast-phase microscopy and confocal microscopy.