Characterization of the metastatic potential of the floating cell component of MIA PaCa-2, a human pancreatic cancer cell line
Characterization of the metastatic potential of the floating cell component of MIA PaCa-2, a human pancreatic cancer cell line
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DOI：
10.1016/j.bbrc.2019.11.120
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发表时间：
2020-02-19
影响因子：
3.1
通讯作者：
Ishiwata, Toshiyuki
中科院分区：
文献类型：
--
作者：
Sasaki, Norihiko;Gomi, Fujiya;Ishiwata, Toshiyuki
在胰腺癌中，形态和功能不同的癌细胞存在于同一患者体内。这种异质性被认为促进了转移和对放化疗的抵抗。Mia Paca-2是已建立的人胰腺导管腺癌(PDAC)细胞系，含有圆形和梭形贴壁细胞以及圆形漂浮细胞。在这项研究中，我们旨在评估与贴壁细胞相比，漂浮细胞是否具有更大的转移潜力和/或对药物诱导的凋亡更具抵抗力。延时分析显示，两种类型的贴壁细胞双向转化，部分贴壁的圆形细胞转化为漂浮细胞。流式细胞仪和电子显微镜显示，大约90%的漂浮细胞是活的。QRT-PCR分析显示，漂浮细胞表达的整合素和三磷酸腺苷结合盒(ABC)转运体水平低于贴壁细胞。相反，除了波形蛋白外，漂浮细胞比贴壁细胞表达更多的上皮细胞到间充质细胞的转化标志。漂浮细胞包括较多的G2/M期细胞，迁移分析显示漂浮细胞相对于贴壁细胞的迁移能力降低。细胞聚集实验表明，漂浮细胞的聚集性低于贴壁细胞。在3D培养中，漂浮细胞来源的球体对包括吉西他滨、5-FU和阿布拉沙尼在内的抗癌药物比来自贴壁细胞的球体更敏感。漂浮细胞来源的球体的干性标志物的表达水平低于贴壁细胞来源的球体。人PDAC细胞株的形态特征可能有助于阐明癌细胞在转移过程中所经历的一系列变化，并可能有助于开发新的PDAC诊断方法和针对PDAC患者的更具体的治疗。(C)2019 Elsevier Inc.保留所有权利。
In pancreatic cancer, morphologically and functionally heterogeneous cancer cells reside within the same patient. The heterogeneity is believed to promote metastasis and resistance to chemoradiotherapy. MIA PaCa-2, an established human pancreatic ductal adenocarcinoma (PDAC) cell line, contains round and spindle-shaped adherent cells, as well as, round floating cells. In this study, we aimed to assess if the floating cells might have greater metastatic potential and/or be more resistant to drug-induced apoptosis compared to adherent cells. Time-lapse analysis revealed that the two types of adherent cells transformed bilaterally, and some of the adherent, round cells converted to floating cells. Flow cytometry and electron microscopy showed that approximately 90% of the floating cells were viable. qRT-PCR analysis revealed that floating cells expressed lower levels of integrins and ATP-binding cassette (ABC) transporters than adherent cells. In contrast, except for vimentin, floating cells expressed more epithelial to mesenchymal transition markers than adherent cells. Floating cells included a larger population of G2/M-phase cells, and migration assays revealed a decreased migration ability by floating cells relative to adherent cells. A cell aggregation assay showed that the aggregative properties of the floating cells were lower than those of the adherent cells. In 3D culture, spheres derived from floating cells were more sensitive to anti-cancer drugs, including gemcitabine, 5-FU, and abraxane, than those derived from adherent cells. Expression levels of stemness markers in the spheres derived from floating cells were lower than those derived from adherent cells. Morphological characterization of human PDAC cell lines may help to clarify the series of alterations cancer cells undergo during the metastatic process and may contribute to the development of new PDAC diagnostics and more patient-specific treatments for those with PDAC. (C) 2019 Elsevier Inc. All rights reserved.