Molecular and Integrative Physiological Sciences, Department of Environmental Health, Harvard School of Public Health, 665 Huntington Avenue, Boston, MA, 02115, USA.
Clin Exp Metastasis. 2013 Mar;30(3):237-50. doi: 10.1007/s10585-012-9531-z. Epub 2012 Sep 8.
We quantified mechanical properties of cancer cells differing in metastatic potential. These cells included normal and H-ras-transformed NIH3T3 fibroblast cells, normal and oncoprotein-overexpressing MCF10A breast cancer cells, and weakly and strongly metastatic cancer cell line pairs originating from human cancers of the skin (A375P and A375SM cells), kidney (SN12C and SN12PM6 cells), prostate (PC3M and PC3MLN4 cells), and bladder (253J and 253JB5 cells). Using magnetic twisting cytometry, cytoskeletal stiffness (g') and internal friction (g″) were measured over a wide frequency range. The dependencies of g' and g″ upon frequency were used to determine the power law exponent x which is a direct measure of cytoskeletal fluidity and quantifies where the cytoskeleton resides along the spectrum of solid-like (x = 1) to fluid-like (x = 2) states. Cytoskeletal fluidity x increased following transformation by H-ras oncogene expression in NIH3T3 cells, overexpression of ErbB2 and 14-3-3-ζ in MCF10A cells, and implantation and growth of PC3M and 253J cells in the prostate and bladder, respectively. Each of these perturbations that had previously been shown to enhance cancer cell motility and invasion are shown here to shift the cytoskeleton towards a more fluid-like state. In contrast, strongly metastatic A375SM and SN12PM6 cells that disseminate by lodging in the microcirculation of peripheral organs had smaller x than did their weakly metastatic cell line pairs A375P and SN12C, respectively. Thus, enhanced hematological dissemination was associated with decreased x and a shift towards a more solid-like cytoskeleton. Taken together, these results are consistent with the notion that adaptations known to enhance metastatic ability in cancer cell lines define a spectrum of fluid-like versus solid-like states, and the position of the cancer cell within this spectrum may be a determinant of cancer progression.
我们定量分析了转移潜能不同的癌细胞的力学特性。这些细胞包括正常和 H-ras 转化的 NIH3T3 成纤维细胞、正常和过表达癌蛋白的 MCF10A 乳腺癌细胞,以及源自人类皮肤癌(A375P 和 A375SM 细胞)、肾癌(SN12C 和 SN12PM6 细胞)、前列腺癌(PC3M 和 PC3MLN4 细胞)和膀胱癌(253J 和 253JB5 细胞)的弱转移性和强转移性癌细胞系。利用磁扭转向量测定法,在较宽的频率范围内测量了细胞骨架的硬度(g')和内摩擦(g″)。g'和 g″随频率的依赖性用于确定幂律指数 x,x 是细胞骨架流动性的直接度量,并量化了细胞骨架在固体样(x=1)到液体样(x=2)状态谱中的位置。NIH3T3 细胞中 H-ras 癌基因表达的转化、MCF10A 细胞中 ErbB2 和 14-3-3-ζ 的过表达以及 PC3M 和 253J 细胞在前列腺和膀胱中的植入和生长,均导致细胞骨架流动性 x 增加。这些先前已被证明能增强癌细胞迁移和侵袭能力的扰动,都使细胞骨架向更类似液体的状态转变。相比之下,具有较强转移性的 A375SM 和 SN12PM6 细胞通过在周围器官的微循环中驻留而扩散,其 x 比弱转移性细胞系对 A375P 和 SN12C 分别小。因此,增强的血液传播与 x 的减小以及向更类似固体的细胞骨架的转变有关。总之,这些结果与这样的观点一致,即已知能增强癌细胞系转移能力的适应性定义了一个类似液体与类似固体的状态谱,而癌细胞在该谱中的位置可能是癌症进展的决定因素。