Distinct Subpopulations of Head and Neck Cancer Cells with Differential Intracellular ROS Exhibiting Diverse Chemoresistance, Stemness and Proliferative Activity

Head and neck squamous cell carcinoma (HNSCC) is a lethal cancer. Emerging evidence supports cancer-initiating cells (CICs) are responsible for tumor growth, chemoresistance, etc. However, the physiological mechanisms by which to maintain the characteristics of CICs in HNSCC (HN-CICs) remain elusive. For hematopoietic stem cells, low intracellular reactive oxygen species (ROSLow) level helps sustain their stemness properties. Therefore; to understand the role of ROSLow cells, which are speculated to be the HN-CICs, may reveal physiological dependencies for head and neck tumorigenesis. Previously, we have identified the existence of HN-CICs from sphere cells. Herein, we initially demonstrated that there were more ROSLow cells in sphere cells than in parental HNSCCs. Further, the ROSLow cells mostly co-expressed CICs surface markers (memGrp78, Glut3, etc.). Next, we exploited flow cytometry to sort the ROSHigh, ROSMidi and ROSLow cells. The isolated ROSLow cells exerted more CICs properties, quiescence, chemoresistance, in vitro malignancy and in vivo tumorigenicity; however, the ROSHigh cells were the most proliferative. Lastly, the pharmacological depletion of ROS modulators in cisplatin-treated HN-CICs was performed to reduce the CICs properties by enhancing cell differentiation and to sensitize the cisplatin cytotoxicity by promoting cell death. Overall, we identified the subpopulations of cells with differential intracellular ROS level harboring diverse chemoresistance, proliferative activity and stemness properties. We also demonstrated that the ROSLow cells not only exert CICs properties but mainly contribute to tumor growth and chemoresistance in HNSCCs. Modulation of low intracellular ROS level should be a future alternative HNSCC treatment in combination with conventional chemotherapy.