Li Yanchun, Chen Jv, Liu Yutong, Zhang Weige, He Wenhui, Xu Hanying, Liu Lianqing, Ma Enlong
Department of Pharmacology, Shenyang Pharmaceutical University, Shenyang, China.
State Key Laboratory of Robotics, Shenyang Institute of Automation, China Academy of Sciences, Shenyang, China.
PLoS One. 2017 Jun 19;12(6):e0179115. doi: 10.1371/journal.pone.0179115. eCollection 2017.
As an inhibitor of microtubule assembly, combretastatin A-4 (CA-4)-induced biological responses in tumor cells have been well known, but the corresponding changes in nano-biophysical properties were not investigated given the lack of an ideal tool. Using AFM technique, we investigated the alteration of nano-biophysical properties when CA-4-treated tumor cells underwent the different biological processes, including cell cycle arrest, apoptosis and autophagy. We found that CA-4-resistant cells were rougher with the presence of characteristic "ridges", indicating that the development of "ridge" structure may be a determinant of the sensitivity of cells to CA-4 compounds. CA-4 induced G2/M arrest and apoptosis in sensitive cells but triggered anti-apoptotic autophagy in resistant cells. CA-4 treatment caused an increase in stiffness in both sensitive and resistant cells. However, these cells exhibited different changes in cell surface roughness. CA-4 decreased Ra and Rq values in sensitive cells but increased these values in resistant cells. The reorganization of F-actin might contribute to the different changes of nano-biophysical properties in CA-4-sensitive and-resistant cells. Our results suggest that cellular nano-biophysical properties, such as "ridges", roughness and stiffness, could be applied as potential biomarkers for evaluating CA-4 compounds, and knowledge regarding how biological alterations cause changes in cellular nano-biophysical properties is helpful to develop a new high-resolution screening tool for anti-tumor agents.
作为一种微管组装抑制剂，藤黄酸A - 4（CA - 4）在肿瘤细胞中诱导的生物学反应已广为人知，但由于缺乏理想工具，其纳米生物物理性质的相应变化尚未得到研究。利用原子力显微镜（AFM）技术，我们研究了CA - 4处理的肿瘤细胞在经历不同生物学过程（包括细胞周期停滞、凋亡和自噬）时纳米生物物理性质的改变。我们发现，具有特征性“脊”的CA - 4耐药细胞更粗糙，这表明“脊”结构的形成可能是细胞对CA - 4化合物敏感性的决定因素。CA - 4在敏感细胞中诱导G2/M期停滞和凋亡，但在耐药细胞中引发抗凋亡自噬。CA - 4处理使敏感细胞和耐药细胞的硬度均增加。然而，这些细胞在细胞表面粗糙度方面表现出不同的变化。CA - 4降低了敏感细胞的Ra和Rq值，但增加了耐药细胞的这些值。F - 肌动蛋白的重组可能导致CA - 4敏感和耐药细胞中纳米生物物理性质的不同变化。我们的结果表明，细胞的纳米生物物理性质，如“脊”、粗糙度和硬度，可作为评估CA - 4化合物的潜在生物标志物，并且了解生物学改变如何引起细胞纳米生物物理性质的变化有助于开发一种新的用于抗肿瘤药物的高分辨率筛选工具。
