Quantitative Evaluation of Cancer Cell Adhesion to Self-Assembled Monolayer-Patterned Substrates by Reflection Interference Contrast Microscopy
Quantitative Evaluation of Cancer Cell Adhesion to Self-Assembled Monolayer-Patterned Substrates by Reflection Interference Contrast Microscopy
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DOI：
10.1021/acs.jpcb.5b11870
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发表时间：
2016-02-25
影响因子：
3.3
通讯作者：
Yoshikawa, Hiroshi Y.
中科院分区：
文献类型：
--
作者：
Matsuzaki, Takahisa;Ito, Kosaku;Yoshikawa, Hiroshi Y.
通过使用自组装单层（SAM）图案化基底和反射干涉衬度显微镜（RICM），已经定量评估了具有不同转移潜能和抗癌药物抗性的癌细胞的粘附。具有优化直径的细胞粘附SAM点可以防止细胞粘附，从而允许通过RICM对单个癌细胞与基底之间的接触面积的统计学可靠数量进行系统评价。统计图像分析显示，高转移小鼠黑色素瘤细胞比低转移细胞表现出更大的接触面积。我们还发现，随着（-）-表没食子儿茶素没食子酸酯（EGCG）浓度的增加，这两种癌细胞类型都表现出从“强”粘附状态到“弱”粘附状态的明显转变，所述（-）-表没食子儿茶素没食子酸酯已知表现出癌症预防活性。粘附转变的数学分析表明，粘附的高转移性小鼠黑色素瘤细胞表现出更多的EGCG耐受性比低转移性细胞。此外，延时RICM观察显示，EGCG以逐步的方式削弱癌细胞粘附，可能是通过粘着斑复合物。这些结果清楚地表明，接触面积可以用作确定癌症表型及其耐药性的定量测量，这将为癌症转移和癌症预防机制提供物理见解。
Adhesion of cancer cells with different metastatic potential and anticancer drug resistance has been quantitatively evaluated by using self-assembled monolayer (SAM)-patterned substrates and reflection interference contrast microscopy (RICM). Cell-adhesive SAM spots with optimized diameter could prevent cell cell adhesion and thus allowed the systematic evaluation of statistically reliable numbers of contact area between single cancer cells and substrates by RICM. The statistical image analysis revealed that highly metastatic mouse melanoma cells showed larger contact area than lowly metastatic cells. We also found that both cancer cell types exhibited distinct transition from the "strong" to "weak" adhesion states with increase in the concentration of (-)-epigallo-catechin gallate (EGCG), which is known to exhibit cancer preventive activity. Mathematical analysis of the adhesion transition revealed that adhesion of the highly metastatic mouse melanoma cells showed more EGCG tolerance than that of lowly metastatic cells. Moreover, time-lapse RICM observation revealed that EGCG weakened cancer cell adhesion in a stepwise manner, probably via focal adhesion complex. These results clearly indicate that contact area can be used as a quantitative measure for the determination of cancer phenotypes and their drug resistance, which will provide physical insights into the mechanism of cancer metastasis and cancer prevention.