Kim Hyonchol, Yamagishi Ayana, Imaizumi Miku, Onomura Yui, Nagasaki Akira, Miyagi Yohei, Okada Tomoko, Nakamura Chikashi
Biomedical Research Institute, National Institute of Advanced Industrial Science Technology, 1-1-1 Higashi, Tsukuba, Ibaraki, Japan.
Biomedical Research Institute, National Institute of Advanced Industrial Science Technology, 1-1-1 Higashi, Tsukuba, Ibaraki, Japan.
Colloids Surf B Biointerfaces. 2017 Jul 1;155:366-372. doi: 10.1016/j.colsurfb.2017.04.039. Epub 2017 Apr 23.
Intercellular adhesion between a macrophage and cancer cells was quantitatively measured using atomic force microscopy (AFM). Cup-shaped metal hemispheres were fabricated using polystyrene particles as a template, and a cup was attached to the apex of the AFM cantilever. The cup-attached AFM chip (cup-chip) approached a murine macrophage cell (J774.2), the cell was captured on the inner concave of the cup, and picked up by withdrawing the cup-chip from the substrate. The cell-attached chip was advanced towards a murine breast cancer cell (FP10SC2), and intercellular adhesion between the two cells was quantitatively measured. To compare cell adhesion strength, the work required to separate two adhered cells (separation work) was used as a parameter. Separation work was almost 2-fold larger between a J774.2 cell and FP10SC2 cell than between J774.2 cell and three additional different cancer cells (4T1E, MAT-LyLu, and U-2OS), two FP10SC2 cells, or two J774.2 cells. FP10SC2 was established from 4T1E as a highly metastatic cell line, indicates separation work increased as the malignancy of cancer cells became higher. One possible explanation of the strong adhesion of macrophages to cancer cells observed in this study is that the measurement condition mimicked the microenvironment of tumor-associated macrophages (TAMs) in vivo, and J774.2 cells strongly expressed CD204, which is a marker of TAMs. The results of the present study, which were obtained by measuring cell adhesion strength quantitatively, indicate that the fabricated cup-chip is a useful tool for measuring intercellular adhesion easily and quantitatively.
利用原子力显微镜（AFM）对巨噬细胞与癌细胞之间的细胞间黏附进行了定量测量。以聚苯乙烯颗粒为模板制作杯状金属半球，并将杯子附着在AFM悬臂的顶端。将附着有杯子的AFM芯片（杯芯片）靠近小鼠巨噬细胞（J774.2），细胞被捕获在杯子的内凹处，然后通过将杯芯片从基质上撤回将其捡起。将附着有细胞的芯片移向小鼠乳腺癌细胞（FP10SC2），并对两个细胞之间的细胞间黏附进行定量测量。为了比较细胞黏附强度，将分离两个黏附细胞所需的功（分离功）用作参数。J774.2细胞与FP10SC2细胞之间的分离功几乎是J774.2细胞与另外三种不同癌细胞（4T1E、MAT-LyLu和U-2OS）、两个FP10SC2细胞或两个J774.2细胞之间分离功的两倍。FP10SC2是从4T1E建立的高转移细胞系，表明随着癌细胞恶性程度的增加，分离功也增加。本研究中观察到的巨噬细胞与癌细胞之间强黏附的一个可能解释是，测量条件模拟了体内肿瘤相关巨噬细胞（TAM）的微环境，并且J774.2细胞强烈表达TAM的标志物CD204。本研究通过定量测量细胞黏附强度获得的结果表明，制作的杯芯片是一种易于且定量测量细胞间黏附的有用工具。
