Abstract
High levels of intracellular reactive oxygen species (ROS) in cells is recognized as one of the major causes of cancer cell apoptosis and has been developed into a promising therapeutic strategy for cancer therapy. However, whether apoptosis associated biophysical properties of cancer cells are related to intracellular ROS functions is still unclear. Here, for the first time, we determined the changes of biophysical properties associated with the ROS-mediated oesophageal cancer KYSE-150 cell apoptosis using high resolution atomic force microscopy (AFM). Oridonin was proved to induce ROS-mediated KYSE-150 cell apoptosis in a dose dependent manner, which could be reversed by N-acetylcysteine (NAC) pretreatment. Based on AFM imaging, the morphological damage and ultrastructural changes of KYSE-150 cells were found to be closely associated with ROS-mediated oridonin-induced KYSE-150 cell apoptosis. The changes of cell stiffness determined by AFM force measurement also demonstrated ROS-dependent changes in oridonin induced KYSE-150 cell apoptosis. Our findings not only provided new insights into the anticancer effects of oridonin, but also highlighted the use of AFM as a qualitative and quantitative nanotool to detect ROS-mediated cancer cell apoptosis based on cell biophysical properties, providing novel information of the roles of ROS in cancer cell apoptosis at nanoscale.
Highlights
Reactive oxygen species (ROS) within cells, such as hydrogen peroxide, superoxide anions and hydroxyl radicals, act as second messengers in the regulation of many important cellular events, PLOS ONE | DOI:10.1371/journal.pone.0140935 October 23, 2015atomic force microscopy (AFM) Analysis of ROS-Mediated Oridonin-Induced KYSE-150 Cell Apoptosis including transcription factor activation, gene expression and cellular proliferation, differentiation and senescence [1]
In the present study, using high resolution AFM, we systematically investigated the biophysical properties of human oesophageal cancer KYSE-150 cells, which were found to be closely related to ROS-mediated apoptosis induced by oridonin
We determined the ROS level in KYSE-150 cells, which demonstrated that oridonin treatment could significantly induce the over-production of ROS in a dose dependent manner and the over-produced ROS induced by oridonin could be eliminated by the pre-treatment of cells with ROS scavenger NAC (Fig 1B)
Summary
Reactive oxygen species (ROS) within cells, such as hydrogen peroxide, superoxide anions and hydroxyl radicals, act as second messengers in the regulation of many important cellular events, PLOS ONE | DOI:10.1371/journal.pone.0140935 October 23, 2015. AFM has been introduced for the study of cancer cell death induced by drug treatment, which provides the high resolution morphological information, and highlights the biomechanical changes during cell death [16,17,18] These works demonstrate that AFM is very useful for the study of anticancer effects of drugs based on the cellular biophysical properties. In the present study, using high resolution AFM, we systematically investigated the biophysical properties of human oesophageal cancer KYSE-150 cells, which were found to be closely related to ROS-mediated apoptosis induced by oridonin. We found that, for the first time, AFM detected morphological damage, membrane ultrastructural changes and biomechanical changes of apoptotic KYSE150 cells induced by oridonin were closely related to the intracellular ROS level of KYSE-150 cells, demonstrating the use of AFM as a powerful nanotool for the study of ROS-mediated cancer cell apoptosis
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